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ECKERT
ANIMAL
PHYSIOLOGY
MECHANISMS AND ADAPTATIONS
DAVID R A N D A L L
U N I V E A S ~O
T FW B R I T I S HC O L U M B I A
1 WARREN B U R G G R E N
KATHLEEN FRENCH
U N I V E R S I TOYF C A L I F O R N I A S, A N D L E G O
W I T H C O N T R I B U T I O NBSY
RUSSELL FERNALD
S T A N F O R DU N I V E R S I T Y
Copyright O 1978, 1983, 1988, 1997 by W. H. Freeman and Company. All rights
reserved.
I
i CONTENTS
I
I
I
PEDAGOGY
The ideas developed in the text are illuminated and
.augmented by liberal use of illustrations and figure
E ckert Animal Physiology has benefitted greatly from
the contributions of several people whose efforts
we gratefully acknowledge. Russell Fernald, Stanford Uni-
Bill Kristan, University of California at Sun Diego
Paul Lennard, Emory University
Jon E. Levine, Northwestern University
versity, participated in the planning and reorganization Harvey B. Lillywhite, University of Florida, Gainesville
of the book and the initial revision of many of the chap- Duane R. McPherson, SUNY at Geneseo
ters. Lawrence C. Rome, University of Pennsylvania, Duncan S. MacKenzie, Texas A&M University
wrote the initial draft of Chapter 10 (Muscles and Eric Mundall, late, University of California at
Animal Movement), which contained most of the updated Los Angeles
material except for the sections on cardiac and smooth Kenneth Nagy, University of California at Los Angeles
muscle and on neuronal control. Harold Atwood, Richard A. Nyhoff, Calvin College
University of Toronto, was involved in some early Richard W. Olsen, UCLA School of Medicine
discusions of the revision. Further, we are grateful for C. Leo Ortiz, University of California at Santa Cruz
the informal comments of associates around the world, Harry Peery, University of Toronto
and for the formal reviews of manuscript, which were J. Larry Renfro, University of Connecticut
provided by the follow~ngcolleagues from across the Marc M. Roy, Beloit College
country: Roland Roy, Brain Research Institute, UCLA School of
Medicine
Joseph Bastian, University of Oklahoma Jonathon Scholey, University of California at Davis
Robert B. Barlow, Institute for Sensory Research, C. Eugene Settle, University of Arizona
Syracuse University Michael P. Sheetz, Washington University School o f
Francisco Bezanilla, UCLA School of Medicine- Medicine
Center for the Health Sciences Gregory Snyder, University of Colorado at Boulder
Phillip Brownell, Oregon State University Joe Henry Steinbach, Washington University School o f
Richard Bruch, Louisiana State University Medicine
Wayne W. Carley, National Association of Biology, Curt Swanson, Wayne State University
Teachers Malcolm H . Taylor, University of Delaware-School o f
Ingrith Deyrup-Olsen, University of Washington Life and Health Sciences
' Dale Erskine, Lebanon Valley College Ulrich A. Walker, Columbia University-College o f
A. Verdi Farmanfarmaian, Rutgers University Physicians
Robert Full, University of California at Berkeley Eric P. Widmaier, Boston University
Carl Gans, University of Michigan Andrea H. Worthington, Siena College
Edwin R. Griff, University of Cincinnati Ernest M . Wright, UCLA School of Medicine-Center
Kimberly Hammond, University of California at for the Health Sciences
Riverside
David F. Hanes, Sonoma State University Finally, the good sense and kind words of Kay Ueno and
Michael Hedrick, California State University- Hayward Kate Ahr, our editors, have much improved the book and
James W. Hicks, University of California at Irvine ensured its publication.
Sara M . Hiebert, Swarthmore College DAVIDRANDALL
William H. Karasov, University of Wisconsin at Madison WARREN BURGGREN
Mark Konishi, California Institute of Technology KATHLEEN FRENCH
xvii
0 ur knowledge of animal physiology is based on infor-
mation (data) derived from experimentation. Since
the ultimate goal of animal physiology is to understand
data is that water-breathing animals regulate acid-base bal-
ance by modifying the excretion of HCO,- in exhaled wa-
ter, while air-breathing animals regulate acid-base balance
how a process operates within an organism, experiments by modifying the elimination of CO, gas in exhaled air. The
must be designed to allow the measurement of key vari- following testable hypothesis could be derived from this
ables (e.g., metabolic rate, blood flow, urine production, general law: A transition from H C O , elimination to CO,
muscle contraction) in the animal (or its cells or tissues) elimination occurs when water-breathing tadpoles meta-
while it is in a known state such as resting, exercising, di- morphose into air-breathing frogs. Although hypotheses
gesting, or sleeping. This kind of experimentation is partic- are framed as statements rather than as questions, the goal
ularly challenging and requires the use of a variety of tech- of experimentation is to test the validity of hypotheses, and
niques and methods. Many of the experimental techniques thus answer the implied questions.
and measuring devices common in animal physiology are Physiological experiments should begin with a well-
"time-honored." These include pressure transducers to formed, specific hypothesis that focuses on a particular
measure pressure, catheter implantation to draw blood or level of analysis and is amenable to a verifiable experimen-
inject samples, respirometers for determining metabolic tal approach. Although a hypothesis such as killer whales
rates, and numerous others. A description of each of these have a very high cardiac output while in pursuit of seals
is beyond the scope of this chapter, especially since such may be interesting and in fact true, it is merely an intellec-
fundamental techniques are well described in texts such as tual exercise to suggest this hypothesis unless a feasible ex-
J. N. Cameron's Principles of Physiological Measurement. perimental approach exists for gathering data necessary to
In this chapter, we will focus on a few of the many molec- accept (prove) or reject (disprove) it. However, the search
ular and cellular techniques that have recently been added for means to test novel hypotheses has been an important
to the physiologist's tool box, briefly describing them and stimulus for development of new experimental techniques
illustrating their use in physiological research. First, how- and measuring instruments. For example, telemetry devices
ever, we consider the nature of hypotheses and the general currently available for gathering data on blood flow in
principles that apply in testing them. small to medium-sized animals like ducks, fishes, and seals
By knowing why and how experiments in animal phys- are being modified for use on even larger animals.
iology are performed-whether they employ traditional or
emerging methods-you will be much better able to eval- The August Krogh Principle
uate the strengths and limitations of the information you August Krogh was a Danish animal physiologist with ex-
will learn in this book. tremely broad interests in comparative physiology. Dozens \
00 @ Q @
000 @@
Spleen cells Myeloma cells
Grow in
mass culture
Antibody Antibody
certainly lead to better methods of controlling or even cur- normal gene product helped alleviate most of the symp-
ing them. Over the last several years numerous laboratories toms of cystic fibrosis in the treated patients.
worldwide have been engaged in a massive project to "map"
the locations of all human genes on the long strands of DNA
in human chromosomes and determine their nucleotide se-
quences. This Human Genome Project is providing invalu-
able data for researchers studying genetic diseases.
DNA cloning and recombinant DNA technology also
form the basis for gene therapy. In this approach to treat-
ing those with genetic disorders, the normal form of the
gene that is missing or defective is introduced into patients.
For example, persons with cystic fibrosis have a defective
CFTR gene and thus cannot produce the normal protein
encoded by this gene. One result of this defect is production
of a very thick mucus in the lungs' airways, which leads to
potentially lethal breathing problems. Molecular biologists
have engineered common cold viruses with the normal
CFTR gene. When some cystic fibrosis patients were in-
fected with an engineered cold virus, the viral particles car-
ried the normal human gene into the patients' lung cells,
where it became established. Subsequent synthesis of the
20 PRINCIPLES OF PHYSIOLOGY
...............................................................................
Plasmid vector mutations that cause a heart with abnormallv thin ven-
tricular walls and another with a constriction of the arte-
DNA fragment rial outflow tract of the heart. Both of these conditions
mimic human disease states.
Mutations often produce abnormal effects only in the
homozygous state (i.e., when an individual receives a mu-
tated form of a gene from each parent). Even when a mu-
tation causes a lethal condition incompatible with long-
term survival, it can be "preserved" in the parents, who are
heterozygous for the mutation, carrying one normal and
one mutated form of the gene. Each time these parents
breed, some of the offspring will be homozygous and show
the abnormal effects. Thus, the heterozygous parents are
a "living gene library" of these mutations.
Bacterial chromosome
Transgenic animals
Transgenic animals are another type of genetically engi-
neered organism with the potential for making great con-
tributions to physiology. A transgenic animal is one whose
genetic constitution has been experimentally altered by the
addition or substitution of genes from other animals of
the same or other species. Transgenic animals (especially,
mice) are at the forefront of the menagerie of animal mod-
els that are helping researchers understand basic physio-
logical processes and the disease states that results from
Figure 2-4 DNA cloning is a way to isolate and maintain individual their dysfunction.
genes. In the cloning procedure illustrated, a specific DNA fragment to Numerous techniques have been employed to produce
be cloned is inserted Into a plasmid vector, whichalso contains a gene transgenic animals. In one method, "foreign" DNA con-
conferring resistance to the antibiotic ampicill~n.When the resulting re-
taining a gene of interest, called a transgene, is injected into
combinant plasm~dsare mixed with E. colicells, a few cells take up a plas-
mid, which can replicate within the cells. If the cells are placed in media
a pronucleus of fertilized eggs (commonly from mice),
containing ampicillin, only those that have taken up the vectorwill grow. which then are implanted into pseudopregnant females. At
As each selected cell multiplies, it eventually forms a colony of cells a relatively low frequency, the transgene is incorporated
(clone) all containing the same recombinant plasmid. into the chromosomal DNA of the developing embryos,
leading to offspring that carry the transgene in all their
"Made-to-order" mutants germ-line cells and somatic cells (Figure 2-5). Mice ex-
As mentioned in Chapter 1, mutations are permanent pressing the transgene then are mated to produce a trans-
changes in the nucleotide sequence of DNA. Mutations, genic line. This approach is used to add functional genes,
which can occur spontaneously or be induced experimen- either extra copies of a gene already present in the animal
tally, are duplicated and passed on to daughter cells at the or a gene not normally present, leading to overexpression
time of cell division. Mutant genes can tell us a great deal of the gene product. Subsequent analysis of the morphol-
about how physiological processes work. The specific dis- ogy and physiology of the transgenic animals can provide
ruption in a physiological process resulting from a single considerable insight into physiological processes that can-
mutant gene can pinpolnt the functions controlled by par- not easily be investigated in other ways.
ticular genes, information that may not be revealed by con- Transgenic animals characterized by underexpression
ventional physiological techniques. or complete lack of expression of a particular gene can be
For example, cardlovascular physiologists are produc- equally informative. M. R. Capecchi (1994)has reviewed
ing and analyzing the effects of mutations in zebrafish to a procedure for replacing a functional gene with a defective
understand heart development. In research described by one, thereby producing so-called knockout mice. These
J-N. Chen and M. Fishman (1997),dozens of specific car- mice cannot express the protein originally coded for by the
diovascular mutations have been produced in zebrafish. replaced gene and thus lack the functions mediated by the
The process starts when adult zebrafish are exposed to missing protein. The molecular and genetic basis of physi-
powerful mutagens-compounds that produce perma- ological processes can be determined by examining the ef-
nent mutations in the germ cell line. Subsequent matings fect of such functional ablation of genes. Knockout mice
of the F, and F2 generations lead to embryos with large are used extensively to unravel human physiological
numbers of mutations. Very rarely, an embryo will appear processes, because human and mouse genes are greater
with just one specific mutation in a structure or process of than 98% identical. To cite a couple of examples, re-
interest. The Fishman group, for instance, has identified searchers are investigating the normal genes that regulate
EXPERIMENTAL METHODS FOR EXPLORING PHYSIOLOGY 21
...............................................................................
individual cells, are used to measure various properties of
cells or inject materials into them. Although cellular phys-
iologists employ these devices in a variety of ways, the
technology used to make them is decades old. Essentially,
I Fertilized eggs collected
from female
a region in the middle of a glass capillary tube is heated to
the point of melting. The ends of the tube are then pulled
apart, which draws the soft spot in the glass down to an
invisibly small diameter before it breaks and separates.
Two micropipettes, each with a drawn-down tip as small
as just a micron in diameter, are produced as a result.
When a micropipette is filled with an appropriate solu-
I Injected eggs implanted
into oviduct of new female
tion, it can function as a microelectrode. Typically, a mi-
cropipette (or microelectrode) is mounted in a microma-
nipulator, a mechanical device that holds the pipette
steady and allows its tip to be moved incrementally in
three different planes.
Measuring electrzcalproperties
Since neurons communicate via electrical signals, micro-
electrodes can be used to "eavesdrop" on their communi-
cation by measuring the electrical signals across the cell
membrane and changes In these signals under different con-
ditions. The microelectrodes used to measure the electric
10-30% of offspring contain transgene
potential (voltage)across the cell membrane cause virtually
1 no flow of current from the cell into the electrode. Thus lit-
axQ
tle or no disruption of the nerve cell occurs even as its com-
A microelectrode
munication with neighboring
for recording
cells iselectrical
being detected.
signals from
neurons or muscle cells is made by filling a micropipette
Breed transgenics to ma~ntainDNA in germ line with an ionlc conducting solution (typicallyKCl) and con-
necting it to an appropriate amplifier. A second electrode
Figure 2-5 A transgenic animal is produced by adding or substituting
genes from another animal of the same or different species.To introduce
connected to the amplifier is placed in the fluid or organism
a transgene into mice, cloned "foreign" DNA is injected into fertilized in the vicinity of the first electrode. When the tip of the first
eggs, which then are implanted into a female. A proportionof the viable electrode is pushed through the cell membrane into the cy-
offspring will retain the transgene, which can be maintained in the germ toplasm, it completes an electric circuit whose properties
line by selective breeding. (voltage, current flow) can be measured. Since microelec-
trode recording techniques were introduced in the 1950s,
early heart development in the embryo and the oncogenes our understanding about the electrical activities within a
responsible for some t y p ~ of
s cancer in studies with knock- cell have increased dramatically.
out mice. One of the most revolutionary advances in microelec-
trode recording methodology is patch clamping. With this
technique, the behavior of a single protein molecule con-
CELLULAR TECHNIQUES stituting an ion channel can be recorded in situ (Latin for
Understanding cells and cellular behavior is a goal of many "in its normal place"), as illustrated in Figure 2-6. This
experiments in physiology. With a knowledge of cellular method lies at the heart of the recent explosion of knowl-
behavior and communication, we can begin to understand edge about membranes, including their channels and how
how communities of cells function as tissues, and tissues as they regulate the movement of materials across the mem-
organs. Physiological analysis at the cellular level has been brane (see Chapters 4-6).
pursued most vigorously using several now-standard tech-
niques. In this section we discuss three very common and Measuring ion and gas concentrations
productive cellular techniques: recording with microelec- Specially constructed microelectrodes can be used to probe
trodes, microscopy, and cell culture. the intracellular concentration of common inorganic ions
including H+,Na+, K+, C1- ,Ca2+,and Mg2+.Because cells
Uses of Microelectrodes and Micropipettes use movements of ions across cell membranes to commu-
Many experiments in cellular physiology make use of mi- nicate and to do work, the magnitude, direction, and time
cropipettes or various types of microelectrodes. These tiny course of ion movements provide important information
glass "needles," which can be inserted into tissues or even about certain processes. ~croelectrodesthat measure the
22 PRINCIPLES O F PHYSIOLOGY
...............................................................................
A sides of the ion-exchange barrier in the tip. Proton-selective
Flu~dto conduct microelectrodes are particularly useful for measuring the
pH of blood and other body fluids.
Specimen embedded
in paraffin wax or
plastic resin and mounted
on arm of microtome
Ribbon of thin
'2W"
I-
sections
Reflecting prism
Ribbon of sections on
glass slide, stained
and mounted under
a cover slip Objective lenses
Specmen
>Condenser
lenses
Path of light
3ase with
ight source
Figure 2-7 Specimens are preparedfor light microscopy by cutting them through a.compound light microscope. (B) The compound optical mi-
into thin sections and staining. (A) Cells and tissue removed from living croscopetransrnitslight vertically up through a condenser lens, the spec-
organisms first are fixed to preserve their structure and then cut ~ n t o
thin imen on the slide, an objective lens, and finally the ocular lens in the eye-
sections using a metal or glass knife. These sections are mounted on a piece, from which the specimen is viewed. [Adapted from Lodish et al.,
glass slide, where they can then be stained for subsequent viewing 1995.1
As improvements have been made in the available op- cent-labeled specimens without the need for thin section-
tics, staining techniques also have improved. Many organic ing. In this microscope, the specimen is illuminated with ex-
dyes, originally developed for use in the textile industry, citing light from a focused laser beam, which rapidly scans
were discovered through trial and error to selectively stain different areas of the specimen in a single plane. The light
particular cellular constituents. Some of these dyes stain ac- emitted from that plane is assembled by a computer into a
cording to the charge, such as hematoxylin, which marks composite image. Repeated scanning of a specimen in dif-
negatively charged molecules like DNA, RNA, and acidic ferent planes provides data with which the computer can
proteins. Howevel; the basis of the specificity of many dyes then create serial sections of the fluorescent images. Figure
is not known. 2-9 compares the images obtained with conventional and
Staining with fluorescent-labeled reagents, rather than confocal fluorescence microscopes.
traditional dyes, increases the sensitivity of visualization. Visualization by other types of microscopy depends
Fluorescent molecular labels absorb light of one wave- on the specimen changing one or more properties of the
length and emit it at another, longer wavelength. When a light passing through the tissue on the slide, rather than
specimen treated with a fluorescent reagent is viewed on fixation and staining. Since these methods do not re-
through a fluorescence microscope, only those cells or cel-
lular constituents to which the label has bound are visual-
ized (Figure 2-8). Probably the most common and useful
type of fluorescence microscopy is immunofluorescencemi-
croscopy in which specimens are treated with fluorescent-
labeled monoclonal and polyclonal antibodies. A good ex-
ample of the images obtained with this technique is shown
in Figure 2-2.
Because immunofluorescence microscopy gives poor re-
sults with fixed thin sections, this technique usually is ap-
plied to whole cells. However, the images obtained by stan-
dard fluorescence microscopy of whole cells represent a
supposition of emitted light coming from labeled molecules
located at many depths in the cell. For this reason, the im-
ages often are blurred. The confocal scanning microscope
eliminates this problem, providing sharp images of fluores-
Incident light
> Eyepiece
source
Barrier filter
d Dlchromatlc
filter
Specimen
Figure 2-8 Aspecimen stained with a fluorescent label is viewed through Figure 2-9 Conventional and confocal microscopy provide different im-
a fluorescence microscope, which produces an image only of structures ages of biological material. These photomicrographs are of a lysed mi-
that bind the label-The incident light source is passed through an exc~ter totic fertilized egg from a sea urchin.A fluorescein-tagged antibody was
I filterthat passes blue light (450-490 nm) to provide optimal illumination used to bind an antibody for tubulin, a major structural component of the
for the specimen.The incident light is directed towards the specimen by mitotic spindle. (A) Conventional fluorescence microscopy shows a
a beam-splitting mirror that reflects light below 510 nm downwards but blurred image as a result of fluorescein molecule above and below the
transmits light above 510 nm upwards. The fluorescent signals emitted plane of focus. (B) Confocal microscopy,which detects fluorescence only
from the labeled specimen pass upward through a barrier filter that re- from within the plane of focus, produces a much sharper image of the
moves unwanted fluorescent signals not corresponding to the wave- same sea urchin egg. [From White et al., 1987.1
lengths emitted by the label used to stain the specimen.
E X P E R I M E N T A L M E T H O D S FOR E X P L O R I N G P H Y S I O L O G Y 25
...............................................................................
quire staining, they can be used on living tissue, provided Electron microscopy
it is thin enough to allow sufficient light to pass through. For all imaging devices, the limit of resolution is directly re-
Bright field microscopy (Figure 2-10A) reveals few details lated to the wavelength of the illuminating light. That is, the
compared to phase-contrast microscopy, in which the im- shorter the wavelength of the illumination, the shorter the
age has varying degrees of brightness and directness due minimal distance between two distinguishable objects (i.e.,
to differential light refraction by different components of the greater the resolution). In electron microscopy, a high-
the specimen (Figure 2-10B). In Nomarski microscopy, velocity electron beam, rather than visible light, is used for il-
also called differential-interference contrast microscopr, lumination. Because the wavelength of electron beams is
an illuminating beam of plane polarized light is split into much shorter than that of visible light, electron microscopes
closely parallel beams before it passes through the tissue have much better resolution. Indeed, modern transmission
specimen and the exiting beams are reassembled into a electron microscopes typically have a resolution of 0.5 nm (5
single image. Slight differences in the refractive index or angstroms, A), whereas light microscopes have a resolution
thickness of adjacent parts of the specimen are converted of no less than about 1000 nrn (1pm). Because the effective
into a bright image, if the beams are in phase when they wavelength of an electron beam decreases as its velocity in-
recombine, or a dark image, if they are out of phase. The creases, the limit of resolution of an electron microscope de-
final image gives an illusion of depth to the specimen (Fig- pends on the voltage available to accelerate the illuminating
ure 2-1OC). In dark-field microscopy, light is directed to- electrons.
wards the specimen from the side so the observer sees only The transmission electron microscope forms images by
light scattered from cellular constituents. The image there- sending electrons through a specimen and focusing the re-
fore appears as if the specimen has numerous sources of sulting image on an electron-sensitive fluorescent screen or
light within it. photographic film (Figure 2-11). The electron beam is mod-
In addition to direct viewing through the microscope, ified by magnets, which bring the electrons into alignment
images can be stored electronically after collection by digi- and focuses them on the specimen, much like the condenser
tal or video cameras. With a digital camera, a color image lens in a compound light microscope. Image formation de-
is collected in its entirety on a two-dimensional array of pends on the differential scattering of electrons by different
photosensitive elements. Although digital cameras provide regions of the specimen; scattered electrons cannot be fo-
a very high resolution, the required light intensity can be cused by the objective lenses and thus do not impinge on
high. Alternatively, a video camera, which has lower light the viewing screen. Because the electron beam passes
requirements, can be used to sample the image according to through an unstained sample nearly uniformly, little differ-
a preset scanning pattern. Because of the high light sensi- entiation of its components is possible without staining.
tivity of the video camera, it permits viewing of cells for The most common stains for electron microscopy are salts
long periods without associated light damage. Such image of heavy metals (e.g., osmium, lead, or uranium), which in-
intensification is particularly important for viewing live crease electron scattering. In photographs of an electron
cells that contain fluorescent labels, which can be toxic to microscope image, components stained with such electron-
cells at high light intensities. dense materials appear dark.
Figure 2-10 Different light microscopictechniques give strikingly differ- ens the visual contrast between different regions of the specimen.
ent images. (A) Bright-field image of a cell, typical of that obtained with (C) Nomarski (differential-interferencecontrast) microscopy provides the
an unstained specimen viewed through a compound light microscope, sensation of depth to the image. [Courtesy of Matthew J. Footer.]
exhibits l~ttlecontrast and few details. (B) Phase-contrast image height-
P R I N C I P L E S OF P H Y S I O L O G Y
...................................
The exquisite detail available from the transmission
Tungsten filament
electron microscope can provide important insights into the
structure of biological tissue (Figure 2-12A). Unfortunately,
the size of the specimen that can be examined is very small,
because it must be thin sectioned. Consequently, it is diffi-
cult to develop an understanding of the three-dimensional
structure without the truly tedious procedure of recon-
structing an image from a series of individual sections. De-
Condenser lens
velopment of various techniques for preparing specimens
for transmission electron microscopy have extended the
range of objects that can be visualized and the information
available from images.
The scanning electron microscope, like the transmis-
sion electron microscope, uses electrons rather than pho-
tons to form images of the specimen. However, the scan-
ning electron microscope collects electrons scattered from
the surface of a specially prepared specimen. This instru-
ment provides excellent three-dimensional images of the
surface of cells and tissues, but it cannot reveal features be-
neath the surface (Figure 2-12B). Before examination in a
scanning electron microscope, the specimen is coated in an
extraordinarily thin film of a heavy metal like platinum.
The tissue is then dissolved away with acid, leaving a metal
replica of the tissue's surface, which is viewed in the micro-
scope. Scanning electron microscopes have a resolution of
Viewing screen or
about 10 nm, considerably less than the resolution of trans-
mission electron microscopes.
Cell Culture
Figure 2-11 Electron microscopes share features such as lenses with
compound opt~calmicroscopes, but use an electron beam rather than a The rearing of cells in uitro (Latin for "in glass") in glass
light beam to illuminate the specimen. In a transmission electron micro- or plastic containers is known as cell culture. This tech-
scope, shown here, an image is formed by passing electrons through an
nique has revolutionized our ability to study cells and
object and projecting them onto a fluorescent screen. In a scanning elec-
tron microscope, electrons reflected from the surface of a specimen
the physiological processes they support at the tissue and
coated with a reflect~vemetal film are collected by lenses and viewed on organ level. Historically, explants (small pieces of tissue
a cathode ray tube. removed from a donor animal) were kept alive and grown
in a flask filled with an appropriate mixture of nutrients
and other chemicals. Today, the most common procedure
is to break up (dissociate) small pieces of tissues and
Since air would deflect the focused electron beam aimed then suspend the dissociated cells in a nourishing chemical
at the sample, the specimen, must be held in a vacuum broth in which they grow and divide as separate entities.
during imaging. Specimens must be very well fixed to pre- Successfully growing cells in vitro requires the right
serve their biological structure during viewing in the electron culture medium, the liquid in which the cells are sus-
microscope. Glutaraldehyde is used to covalently cross-link pended. Up until the early 1970s, cells from all animals
proteins and osmium tetroxide to stabilize lipid bilayers. Af- were routinely grown in liquid medium consisting largely
ter fixing, the specimens are infiltrated with a plastic resin. of either serum (a clear component of blood plasma) from
Thin sections cut from the resin block then are stained and horses or fetal calves or of an unrefined chemical extract
finally placed on a metal grid in the transmission electron mi- made from ground-up chick embryos. However, these
croscope. Specimens must be sectioned into extremely thin media were poorly defined chemically, containing nu-
slices (50-100 nm thick) to allow penetration by the elec- merous unidentified compounds. Moreover, it was diffi-
tron beam. Only diamond or glass knives are sharp enough cult to predict whether cells from a particular source
to cut tissue sections into such thin slices. Glass knives are would grow in one of these media, or what components
formed by breaking on the diagonal a 2.5-cm glass square might be added if the first attempt was unsuccessful.
that is about 5 mm thick. Because glass is actually a slow- Growing cells in vitro was largely a matter of trial and er-
moving liquid, the edge formed is only sharp enough to cut ror (and luck). Today, defined culture media manufac-
tissue for a few hours before molecular flow of glass dulls the tured according to precise chemical formulas are avail-
edge. Although extremely expensive, diamond knives do not able for research. However, the successful culture of many
suffer from this problem and thus are the preferred tool for cell types requires addition of a small trace (less than 5%)
cutting thin sections. of horse serum to such defined media. This observation
tion, many types of animal cells have not yet been suc-
cessfully cultured. However, the list of cells that can be
cultured is constantly growing, the result of refinements
in media and culture techniques. For example, cell strains
derived from the following tissues and organs can be
grown in culture:
M
In contrast to normal animal cells, cancer cells com-
0.2 pm
monly exhibit rapid, uncontrolled growth in the body and
are c a ~ a b l eof indefinite growth in culture. Treatment of
some normal cultured cells with certain agents may cause
transformation, a process that makes them behave like can-
cer cells that have been isolated from tumors. Such trans-
formed cells also can be cultured indefinitely. Homoge-
neous populations of such "immortal" cells are termed cell
lines. Although normal cells differ from cancer cells and
transformed cells in many ways, cell culture of the latter has
permitted certain types of studies that are not feasible with
primary cell cultures of normal cells.
Cell culture has numerous potential uses in animal
physiology. New devices such as silicon wafer sensors for
measuring acidity and other variables have been combined
with cell culture techniques to provide important insights
into cellular and organismal physiology. For example, the
hormonal regulation of H+secretion from a variety of cells
grown in vitro can be studied by stimulating the cultured
cells with agonists and antagonists and measuring the
changes in the rate of medium acidification. This approach
has also been used to study tissues and organs with unusual
rates or ~ r o ~ e r t iof
1 L
e sH + secretion, such as the swim blad-
der tissues of fishes.
A
condiner sepa;ated
with solvent components
B w ~ t hSDS
Add solvent
through column
Partially
separated
proteins
IStain to visualize
separated bands
Collect fractions;
larger components
pass more quickly
II Proteins
Figure 2-14 Chromatography is a powerful technique for separating the
components of a mixture in solution. (A) In paper chromatography, the
sample is applied to one end of a piece of chromatographic paper and
@ @$ @ @
Ij separated
by size
dried. The paper is then placed into a solution containing two or more
solvents, which flow upward through the paper via capillary action. Dif-
ferent components of the sample move at different rates in the paper be- Figure 2-1 5 Gel electrophoresisseparates the components of a mixture
cause they have different relative solubilities in the solvent mixture. Af- based on their charge and their mass. Proteins commonly are separated
ter several hours, the paper is dried and stained to determine the location by SDS-polyacrylam~de gel electrophoresis, as illustrated here (A) SDS,
and relative amounts of the separated components. (B) In column chro- a negatively charged detergent, is added to the sample to coat the pro-
matography,the sample is applied to the top of a column that contains a teins. (B) The sample then is placed in a well in the polyacrylamide gel
permeable matrix of beads through which a solvent flows. Then solvent and an electricf~eldis applied. Small proteins move more easily and faster
is pumped slowly through the column and is collected in separate tubes along the length of the gel than larger ones. (C)After a period of time,
(calledfractions) as ~temerges from the bottom. Components of the sam- the rotei ins in the mixture are separated into bands composed of
ple travel at d~fferentrates through the column and are thus sorted Into proteins of different sizes. These can be visualized by various protein-
d~fferentfract~ons. staining reagents. [Adapted from Lodish et al., 1995.1
30 PRINCIPLES OF PHYSIOLOGY
Band
corresponding
-
Electric
current
Electrophoretic
transfer
reagent spec~flc
to component A
corresponding
t o component B
Gel after Polymer Audioradiograms
electrophoresis sheet
Figure 2-16 Southern, Northern, and Western blotting are similar proceduresfor separating and identifying specific DNA fragments, mRNAs, and pro-
teins, respectively,within a mixture. In each method, the components of a sample first are separated by gel electrophoresis; the separate bands are trans-
ferred to a polymer sheet, which then is flooded w~tha radiolabeled reagent specific for the component of interest. The presence and in some cases
the quantity of the labeled component is determined by autoradiography.See Table 2-1 for details of each procedure.
within a complex mixture can be detected by Western blot- be the deflection of the ions from a standard path as they
ting, also known as irnrnunoblotting. (As yet, there are no head towards the analyzer. The degree of deflection is de-
Eastern, South-Western, etc., blots, but it is probably just a tected by an array of detectors, which then allows deter-
matter of time.) Table 2-1 summarizes the unique features mination of the presence and quantity of gases in the gas
of the three blotting procedures. sample introduced into the mass spectrometer.
Many of the common methods of determining compo- The various techniques for measuring chemical com-
sition are applicable to solutions but not gases. The mass position described in this section are widely used by physi-
spectrometer, however, can distinguish the different gases ologists, but many others also are employed in physiologi-
composing a gaseous mixture based on their mass and cal research. To learn about additional methods and further
charge. Animal physiologists most often use this instrument details about those discussed here, you can refer to chem-
to determine the composition of respiratory gases while an istry and biochemistry textbooks.
animal is resting or exercising in an experimental setting.
Figure 2-17 illustrates the basic design of a mass spectrom- Measuring Concentration: How Much Is Present
eter. The gas sample first is ionized by intense heating and Most instruments or analytical techniques used to deter-
passage through an electron beam. The charged ions then mine the composition of a fluid or gas mixture also provide
are focused and accelerated by an electric field into an an- data about the concentration of the components present.
alyzer where the beam of ions is deflected by either an ap- For example, the degree of color change produced in a col-
plied magnetic field or by passage through tuned rods emit- orimetric assay depends on how much of the substance be-
ting specific radio frequencies that will deflect ions. The ing measured is present in the sample. Likewise, the output
lighter the ion mass and smaller its charge, the smaller will signal from a mass spectrometer depends not only on the
TABLE 2-1
Electrophoreticblotting procedures
- - - -
Southern blottlng ' DNA fragments produced by cleavage of DNA Electrophorese mlxture of dsDNA fragments on agarose
wrth restrlctlon enzymes or polyacrylamlde gel, denature separated fragments
Into ssDNA and transfer bands t o polymer sheet, use radlo-
% labeled ssDNA or RNA t o label fragment of Interest, detect
labeled band wlth autorad~ography
Northern blotting Messenger RNAs Denature sample mixture; electrophorese on polyacrylamide
gel and transfer separated bands t o polymer sheet; use radio-
labeled ssDNA t o label mRNA of Interest; detect labeled band
with autoradiography
Western blotting Proteins Electrophorese sample mixture on SDS-polyacrylamide gel
and transfer separated bands t o polymer sheet; use radio-
labeled monoclonal antibody t o label protein of interest; de-
tect labeled band with autoradiographyt
+Ifradiolabeled monoclonal antibody is not available, then the band contaming the antibody-protein complexes can be detected by addlng a secondary
antibody that binds to any monoclonal antibody Th~ssecondary antibody is covalently linked to an enzyme, such as alkaline phosphatase, that catalyzes
a colorimetric reaction. When substrate is added, a colored product forms over the band with the protein of interest, generating a visible colored stain
in this region of the blot.
Applied magnetic field
I ,,, To vacuum
Accelerator
1 beam To amplifier
ion~zation and recorder
Sample
B py$:.~Fs fi
particles
Ion Ion Ion Ion Ion Ion
source collector source collector source collector
Figure 2-17 The identity of gases in a mixture and their concentrations flow in a curved path (3). Finally, the ion beam is detected using an ion col-
can be determined by mass spectrometry. (A) The fixed-collector mass lector situated at the end of the analyzer tube (4).The extent to which an
spectrometer, which detects how much an ionized sample is deflected by ion is bent in the applied magnetic field depends on the strength of the
an imposed magnetic field, has four essential parts. First is a carefully con- field and the ion's mass, charge, and velocity. Only those Ion species that
structed inlet device (I) through which the sample is delivered to the sys- are bent so their ~ a t h s
parallel the sides of analyzer tube will reach the ion
tem with a constant viscous flow. Second is an ionization chamber (Z), collectors and be detected. (B) At a constant magnetic field, the specific
kept under vacuum and at high temperature (about 19O0C),where the type of particles detected in the mass spectrometer is determined by the
sample passes through an electron beam and is accelerated vla applica- strength of the ion~zation voltage, which can be varied. The lower the ion-
tion of an electric field. The gas molecules leave this chamber as nega- izing voltage, the heavier is the particle detected. [Adapted from Fes-
tively charged ions. Third is an analyzer tube in which the accelerated senden and Fessenden, 1982.1
beam of ions are subjected to a magnetic field that causes the ions to
types of gases present in a mixture, but how much of each difficult, if not impossible, to do by studying the intact
is present. Thus the output signal produced by an analytical organs in situ; instead, experiments are conducted on iso-
instrument- be it a transmission spectrophotometer, den- lated organs removed from the animal by surgery and
sitometer, or mass spectrometer-is directly related to the maintained in an in uitro artificial environment. Two ex-
concentration of the substance responsible for the signal. amples will illustrate the power of this experimental
Typically, the analytical technique being used to deter- approach.
mine the concentration of a particular substance is carried When the heart of almost all vertebrates, including
out on several samples of the substance at different known mammals, is isolated and placed in a bath of saline, it con-
concentrations'; the output signals then are plotted against tinues to beat and perform work by pumping saline or
the concentrations, yielding a standard curve. The actual other fluid supplied to it. The isolated vertebrate heart will
concentration of an experimental sample corresponding to continue to beat in the absence o f neural input if it is kept
the output signal it produces is determined by comparison at an appropriate temperature and is perfused with an oxy-
with this standard curve. genated solution that has the correct ionic composition and
contains an energy source such as glucose. With the heart
isolated, physiologists can measure the effect of chemical
EXPERIMENTS WITH ISOLATED stimulation by drugs and hormones or electrical stimula-
ORGANS AND ORGAN SYSTEMS tion of nerves within the heart on the heart rate, amplitude,
All animals have several differentmajor organ systems that flow rate, and mechanical movements. Experiments per-
must be coordinated and controlled to help maintain formed on isolated hearts have been fundamental in ad-
homeostasis. As we'll examine in later chapters, the func- vancing our knowledge of the cardiovascular system.
tions of these organ systems are regulated primarily by A second example is the vertebrate pineal gland, a small
neural and/or hormonal inputs. To understand physiolog- organ found at the top of the vertebrate brain. The pineal,
ical control mechanisms, the key controlling inputs and which plays a key role in regulating daily (circadian)
their sources must be characterized. In many cases, this is rhythms in physiological processes, is sensitive to light-
32 PRINCIPLES O F PHYSIOLOGY
.........................................
related stimuli and releases various amounts of regulatory feeding, two of the most important behaviors engaged in by
chemicals into the bloodstream as a function of the time of any animal; both reproductive and feeding behaviors are
day. When the pineal gland is isolated and placed into an greatly affected by an animal's physiological state. Careful
appropriate culture system, it continues to exhibit a circa- observations usually can reveal which behavioral patterns
dian rhythm. Direct experimentation with this in vitro of one individual influence another and may suggest why
preparation has provided answers to specific questions con- this might be so. For example, in stickleback fish, the dis-
cerning pineal gland regulation of physiological systems. play of a red belly by a male signals to other male stickle-
backs that he is defending a nest and to females that he is
interested in spawning. Thus, the meaning of this signal de-
OBSERVING AND MEASURING pends on the sex of the receiver. The red belly arises from
ANIMAL BEHAVIOR
physiological processes triggered by the onset of the breed-
Scientists studying animal ~ h ~ s i o l frequently
og~ supple- ing season. The coordination between behavior and phys-
ment their experiments with observations of animal be- iology in this species was investigated using behavioral
havior. Useful behavioral experiments are difficult to analysis to guide physiological investigation.
perform, however, because the animals must be in an ap-
propriate physiological state (e.g., breeding, rearing young, Methods in Behavioral Research
digesting a meal, to name a few). Further, the experiment A variety of instruments are used to record and analyze the
must exploit natural behavioral tendencies in the animal. physiological basis of specific behavioral acts. In some ex-
Despite these difficulties, experimental methods to control periments, high-speed video cameras are used in conjunc-
and stimulate specific behavioral states can provide impor- tion with electrophysiological detectors of neural or mus-
tant insights into physiological processes that are not al- cular activity to capture simultaneously both the behavior
ways amenable to direct physiological investigation. The and its physiological underpinnings. Since behavioral acts
prerequisite for such experimentation is a thorough knowl- of interest are often rapid and fleeting, these events typically
edge of the natural behavior of the animal in its habitat. are recorded at high speed and the video tape played back
at slow speed to aid analysis. The use of x-ray cameras al-
The Power of Behavioral Experiments lows examination of the interaction of skeletal components
Research in the 1950s and 1960s on the retrieval behavior during specific behaviors (e.g., feeding, running on a tread-
of ground-nesting birds illustrates how behavioral studies mill). As in so many other aspects of physiology, the avail-
can contribute to physiological knowledge. K. Z. Lorenz ability of inexpensive, fast computers with ever-increasing
and N. Tinbergen discovered that geese not only recognized data storage capabilities has revolutionized the acquisition
their eggs and recovered them if they lay outside the nest, and analysis of data.
but also would retrieve a wide variety of objects (e.g., Figure 2-18 illustrates how many of the commonly
grapefruits, light bulbs, baseballs) laying near their nest. used techniques to study animal behavior and its underly-
Tinbergen and his students subsequently conducted inge- ing physiological processes can be brought to bear on a sin-
nious experiments with gulls in which they offered pairs of gle behavior-the feeding strike of a venomous snake. To
objects to the birds and recorded which one was recovered discover how such a strike proceeds, the motion of the
first. By exploiting the process of painvise comparison, they body and jaws must be related to the forces exerted by con-
could define the properties that gulls use to choose what to traction of the jaw muscles. The rapid strike is recorded in
retrieve. Although the birds would retrieve many different two views, dorsal and lateral, using a video camera view-
objects, these experiments showed that real eggs are always ing the animal directly and via a mirror set at 45' above the
preferred over unnatural objects. The relative size, color, snake. Quantification of the position of the animal is pos-
and speckling of an egg were found to contribute indepen- sible because of a grid image in the background that is in-
dently to the likelihood that an egg would be retrieved. cluded in the video images. The snake is placed on a plat-
Taken together, these experiments revealed that for gulls, form that records the force exerted along three orthogonal
eggs provide a powerful natural stimulus that induces spe- axes. By measuring this set of external parameters, the in-
cialized retrieval behavior. Armed with knowledge of the vestigator can record the forces associated with the snake's
exact properties of the stimuli causing this behavior, phys- movement across the surface. The force exerted by the jaws
iologists have been better able to conduct physiological ex- of the snake are recorded by a strain gauge mounted on the
periments about the nature of vision in birds. head, and muscle activity is measured by electrodes into the
Behavioral experiments often analyze the total time the four lateral jaw muscles. All of the data are recorded on
animal under study spends performing each behavior and both tape and in a computer using data-acquisition hard-
the temporal sequence of behaviors. These data, in con- ware and software.
junction with information about the behavior of other an- The values of the measured variables are typically dis-
imals and key environmental variables, frequently reveal played as a function of time and related to the behavioral
how closely behavior is related to the internal state of the analysis recorded on videotape. Data from such an experi-
animal. The great majority of information about animal be- ment reveals how contraction of the muscles results in posi-
havior collected in this way pertains to reproduction and tioning of the fang and closing of the jaws around the prey.
EXPERIMENTAL METHODS FOR EXPLORING PHYSIOLOGY 33
...............................................................................
Force
platform
Oscilloscope
High-
speed recorder
camera
Tape
recorder
Figure 2-18 The feeding strike of a venomous snake can be analyzed ing skull bones. (B) The snake is placed on a force-recording table and
to determine the muscles used and the pattern in which they contract. videotaped as it strikes its prey. The leads from the electrodes and strain
(A) To record electrical potentials from the jaw musculature, fine bipolar gauge are connected t o electronic amplifiers, which boost their low-
wire electrodes are placed surgically into the four lateral jaw muscles in voltage signals. The amplified signals are displayed on an oscilloscope
a procedure that is performed under anesthesia. Astrain gauge is also at- and chart recorder and stored on a tape recorder and computer.
tached to the top of the snake's head t o measurethe motion of underiy-
.. :.. , .. C H . 4 P T E R
3
MOLECULES, ENERGY,
AND BIOSYNTHESIS
First 1 2
shell H He
Second 3 4 5 6 7 8 9 1 0
shell LI Be B C N O F N e
Thlrd 11 12 13 14 15 16 17 18
shell Na Mg Al Si P S CI Ar
Fovrlh 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36
shell K Ca Sc Ti V Cr Mn Fe Co NI Cu Zn Ga Ge As Se Br Kr
Flfth 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54
shell Rb Sr Y Zr Nb Mo Tc Ru Rh Pd Ag Cd In Sn Sb Te I Xe
Slxth 55 56 57 72 73 74 75 76 77 78 79 80 81 82 83 84 85 86
shell Cs Ba La Hf Ta W Re 0s Ir Pt Au Hg TI Pb BI Po At Rn
Seventh 87 88 89 104 105 106
shell Fr Ra Ac
58 59 60 61 62 63 64 65 66 67 68 69 70 71
Ce Pr Nd Pm Sm Eu Gd Tb Dy Ho Er Tm Yb Lu
90 91 92 93 94 95 96 97 98 99 100 101 102 103
Th Pa U Np Pu Am Cm Bk Cf E3 Fm Md No Lw
Figure 3-1 In the periodic table of the elements, each row corresponds to a different electron orbital shell. The
elements in colored lettering are physiologically important in their ~onicforrns.
\
MOLECULES, ENERGY, A N D BIOSYNTHESIS 39
............................................................................
TABLE 3-1 portant to animal physiology because they dominate inter-
Comparison of the chemical composition of the human body actions among elements central to organic life. Indeed, the
with that of seawater and the Earth's crust*
. - interactions amongst these three particles dictate the at-
Human body Seawater Earth's crust traction among elements necessary for life itself.
Each atom consists of a dense nucleus of protons and
neutrons surrounded by a "cloud" of electrons equal in
number to the protons in the nucleus. The atomic particles
have the following charge and mass (in daltons, Da):
Proton: + 1; 1.672 Da
Neutron: 0; 1.674 Da
Electron: -1; 0.001 Da
Oxygen Fluorine
Phosphorus Chlorine
3 to 10 electrons (e.g., carbon, nitrogen, and oxygen), this its third and outer shell, is less reactive than fluorine, which
first shell is occupied by two electrons; the remaining elec- has seven electrons in its second and outer shell (see Figure
trons occupy a second shell, which can contain up to eight 3-2). Both atoms have a tendency to gain one electron to
electrons, located farther from the nucleus. In atoms with complete the outermost shell, but this tendency is greater in
11to 18 electrons (e.g., sodium, phosphorus, and chloride), fluorine, since its outermost shell feels a stronger electro-
a third shell is formed, which also can accommodate up to static pull from its nucleus than the larger chlorine atom.
eight electrons. The fourth and fifth shells each can hold up As a result, with all other things being equal, a small atom
to 18 electrons (Figure 3-3). forms stronger and hence more stable bonds with other
When the outermost shell of an atom contains the max- atoms than does a large atom.
imal number of electrons possible in that shell-that is,
when it cannot accommodate additional electrons-the
atom is highly stable and resists reactions with other atoms.
This is true of all the noble gases, such as helium and neon,
THE SPECIAL ROLES OF H, 0, N,
which appear at the far right of the periodic table. Most
AND C IN LIFE PROCESSES
elements, however, have incomplete outer electron shells Let's return to Wald's view that hydrogen (H), oxygen
and are therefore reactive with certain other atoms. Hy- (0), carbon (C),and nitrogen (N)dominate the composi-
drogen, for example, has one rather than two electrons in tion of biological systems because they lend themselves es-
its only shell, and oxygen has only six, instead of eight, elec- pecially well to the chemistry of living systems. Examina-
trons in its outer shell. Thus, the hydrogen atom and the tion of the periodic table reveals that these four elements
oxygen atom both have a tendency to share electrons so as have one or two electron shells. Of the other elements
to fill their respective outer shells and bring them into more with only one or two electron shells, helium and neon are
stable configurations. virtually inert, rare gases; boron and fluorine form rela-
Although the number of electrons in the outer shell has tively rare salts; and the metals lithium and beryllium
an important influence on the physical characteristics and form easily dissociated ionic bonds. In contrast, H, 0,N,
reactivity of an atom, other physical features are also im- and C will form strong covalent bonds by sharing one,
portant in determining its chemical properties. One of these two, three, and four electrons, respectively, to complete
is the size (or weight) of the atom. The heavier an atom (i.e., their outer electron shells.
the more protons and neutrons in its nucleus), the more Why are strong bonds important in living systems?
electrons surround the nucleus. As the number of electrons Without strong bonding, subtle changes in temperature,
exceeds ten and a third shell of electrons appears, the va- pH, or other variables in h v i r o n m e n t surrounding a
lence electrons (i.e., those in the outermost shell) are cor- molecule could cause it to break down or rearrange. Con-
respondingly more distant from the compact nucleus, and sider, for example, the biological chaos that would result
hence less strongly attracted by it than the valence electrons if the chemical bonds in the hereditary material formed by
of atoms with only two shells. This is because the electro- DNA were easily dissociated and subject to alterations (i.e.,
static interactions between the single charged electrons and mutations). In fact, mutations are quite rare (less than one
protons (monopoles) diminish with the square of the dis- per gene in every 10,000 replications) because the atoms
tance between them. Thus, chlorine, with seven electrons in composing DNA are strongly bonded to each other in a
multitude of combinations. The short-term integrity of
each organism and each species depends upon the stable
bonds holding together the structures of DNA and other
macromolecules.
Of the four major biologically important elements,
three (0,N, C) are among the very few to form double or
triple bonds. These multiple bonds not only increase the
stability of molecules containing them, but also greatly in-
crease the variety of molecular configurations that can be
formed by reaction of these elements (Figure 3-4). For ex-
ample, oxygen can react with carbon to form carbon diox-
ide, CO,. Since the two double bonds connecting the 0
atoms to the C atom satisfy the tendencies of these atoms
to react, the CO, molecule is relatively inert. Therefore,
CO, can diffuse readily from its source of production to be-
come available for recycling through the photosynthetic
Figure 3-3 Each orbital shell in atomicstructure can accommodate a set
process of green plants. Because the carbon atom has a
maximal number of negatively charged electrons, as shown here for the
first four orbital shells (numbered 1,2, 3, and 4). Atoms with more elec- valence of four, it can form four single bonds, two double
trons than needed to fill, for example, shells 1 and 2, will form an addi- bonds, and combinations of single with double or triple
tional shell or shells. bonds, endowing it with the ability to form a great diver-
MOLECULES, ENERGY, A N D BIOSYNTHESIS 41
.......................................
H WATER: THE UNIQUE SOLVENT
I
H-C-OH C-OH
I I We live on the "water planet." Because water is so com-
O=C=O H-C-OH H,N-C-H mon, it is often regarded with indifference, as some sort of
I I inert filler occupying space in living systems. The truth is
Carbon dioxide H-C-OH H-C-CH,
I I that water is directly and intimately involved in all details
of animal physiology. Water is a highly reactive substance,
Glycerol Valine quite different both physically and chemically from most
other liquids. Water possesses a number of unusual and
special properties of great importance to living systems. In-
c I +/C-S H OH deed, life as we know it would be impossible if water did
H N/ c' -C-N, I I I not have these properties. The first living systems presum-
I I 11 I C=C-C-C-H
ably arose in the aqueous environment of shallow seas. It
H-C-C\ C
,H H I I I
I N H-C-H H H is therefore not surprising that the living organisms of the
present are intimately adapted at the molecular level to the
special properties of water. Today even terrestrial animals
Thiamin (vitamin 8,)
consist of 75% or more of water. Much of their energy
Figure 3-4 The ability of carbon, oxygen, and nitrogen to form double
expenditure and physiological effort is devoted to the con-
bonds (in red), in addition to single bonds, greatly increases the
diversity of molecular structures including these elements. Glycerol is a
servation of body water and the regulation of the chemical
constituent of fats, and valine is one of the amino acids present in natural composition of the internal aqueous environment.
proteins. The special properties of water so important to life stem
directly from its molecular structure. Therefore, we should
begin with a brief consideration of the water molecule.
sity of atomic combinations with itself and other atoms, in- The Water Molecule
cluding straight and branched chains, as well as ring struc- Water molecules are held together by polar covalent bonds
tures (see Figure 3-4). between one 0 and two H atoms. The polarity (i.e.,uneven
Silicon (Si),which is in the same column and just below charge distribution) of the covalent bonds results from the
carbon in the periodic table, has some properties similar to strong tendency of the 0 atom to acquire electrons from
those of carbon. Unlike carbon, however, it is larger and other atoms, such as H. This high electronegativitycauses
does not form double bonds. Therefore, it combines with the electrons of the two H atoms in the water molecule to
two atoms of oxygen by two single bonds only: occupy positions statistically closer to the 0 atom than to
the parent H atoms. The 0-H bond is therefore about
40% ionic in character, and the water molecule has the fol-
lowing partial charge distributions (6 represents the local
partial charge of each atom):
Since the outer electron shells in all three atoms of silicon
dioxide (SiO,) are not filled, the SiO, molecule readily
bonds with others of its kind, forming the huge polymeric
molecules that make up silicate rocks and sand. Thus, it is
evident that silicon, even though it has some properties sim-
ilar to those of carbon, is far more suited for the formation The angle between the two 0-H bonds in the water
of stone than it is for large-scale participation in the orga- molecule, rather than being 90" as predicted for purely co-
nization of biological molecules. valent bonding, is found to be 104.5" (Figure 3-5). The in-
Besides its important role in combining with hydrogen creased angle can be ascribed to the mutual repulsion of the
to form water, oxygen acts as the final electron acceptor in two positively charged H nuclei, which tends to force them
the sequence of oxidation reactions through which chemi- apart. In contrast, the S-H bonds in the hydride of sulfur,
cal energy is released by cell metabolism. This important H,S, are purely covalent; so there is no asymmetrical charge
ability to oxidize (accept electrons from) other atoms and distribution as in H 2 0 . Thus, the bond angle in H2S is
molecules is due to the oxygen atom's incomplete outer closer to 90". Because of the semipolar nature of 0-H
electron shell and relatively low atomic weight. bonds, H 2 0 differs greatly, both chemically and physically,
In addition to the four major biological elements, nu- from H,S and other related hydrides. Why is this?
merous other elements participate in cell chemistry, though The uneven distribution of electrons in the water
in lesser numbers (see Table 3-1). These include phospho- molecule causes it to act like a dipole. That is, it behaves
rus (P) and sulfur (S), the ions of four metallic elements somewhat like a bar magnet, but instead of having two
(Naf, K+, M$+, and Ca2+),and the chloride ion (Cl-). We opposite magnetic poles, it has two opposite electric poles,
will return to these later. positive and negative (see Figure 3-5). As a result, the
Van der Waals
dipole moment
Figure 3-5 In the water molecule, the electron density is greater around
the oxygen atom than around the hydrogen atoms, giving the 0-H
bond a semipolar character. The mutual repulsion between the resulting
partial positive charges on the hydrogen atoms causes the angle be-
tween the two 0-H bonds to be greater than that characteristic of
purely covalent bonds. 6+ and 6- indicate a part~alpositive and negative
charge, respectively.
Wo Ion
Point
charge
Ion
Water d (arbitrary units)
Figure 3-8 Interactions between ions and charged sites are influenced with the square of the distance. For a dlpole such as the water molecule
by the distance separating them. The electrostatic force, f, between an the value of a can be as high as 4.0. (B) The drop in electrostatic force as
ion and a site of opposite charge varies inversely with the distance, d, afunction of distance is illustrated forthese twovalues of a. In the case of
raised to some power, a, between them: f l l d a .(A) For a point charge, water and a positive point charge, the actual value of a is closer to 3.0.
or monopole, the exponent a equals 2.0, so that the force drops inversely
hydrophilic (water-attracting) polar head and a hydropho- logical membranes in living cells. Micelle formation may
bic (water-repelling)nonpolar tail (Figure3-9A). If a mixture have provided the basis for the first cell-like organization of
of water and sodium oleate is shaken, the water will disperse a living system in the organic-rich shallow seas in which life
the latter into minute droplets. The sodium oleate molecules is believed to have undergone its first evolutionary stages.
in such a droplet, or micelle, are arranged with their hy-
drophobic, nonpolar tail groups huddled in the center and PROPERTIES OF SOLUTIONS
their hydrophilic, polar head groups arranged around the As noted already, water plays a critical role in living systems.
perimeter, facing outward, so as to interact with the water Indeed, many of the physical and chemical processes of cells
(Figure 3-9B).The same behavior is exhibited by phospho- occur in water solution. The fluids within the cells and tis-
lipid molecules, which also consist of hydrophobic and hy- sues of animals, as well as the aqueous environment in
drophilic groups. The tendency of amphipathic molecules to which aquatic animals live, are critically influenced by the
form micelles in water is important in the formation of bio- solutes-particularly the electrolytes-that they contain.
Polar head
tail I II
,cH
The concentration of water remains virtually unaltered Note that the pH scale is logarithmic and typically
by its partial dissociation into H + and OH-, because the ranges from 1.0 M H + to 10-l4 M H + (Table 3-3). Thus,
concentration of each of the dissociated products is only a M solution of a strong acid such as HC1, which
M mol .L-I), whereas the molar concentration dissociates completely in water, has a pH of 3.0. A solu-
of water in a liter of pure water (equal to 1000 g) is tion in which [H+] = [OH-] = M has a pH of 7.0,
1000 g .L-I divided by the gram molecular weight of wa- and so forth. A solution with a pH of 7 is said to be neu-
ter (18 g.L-l), or 55.5 M (55.5 mo1.L-I). Equation 3-1 tral-that is, neither acidic nor basic. However, the ratio
can thus be simplified to of [Hf] to [OH-] depends on temperature, so the true
"neutral pH" (called the pN) at which [Hf] = [OH-]
actually rises above 7.0 at temperatures below 25°C
and falls below 7.0 at temperatures above 25°C. The pH
of a solution can be conveniently measured as the voltage
Recall that a consequence of the law of mass action is the
produced by H + diffusing through the proton-selective
reciprocal relation between the concentrations of two com-
glass envelope of an electrode immersed in the solution
pounds in an equilibrium system. This reciprocity is ap-
(Figure 3-12).
parent in the constant [H+][OH-], which may be lumped
with the molarity of water (55.5) into a constant that will
be termed the ion product of water, Kw. At 25°C this has
a value of 1 x 10-14:
Human gastric
acidity
1 3
4
10-~
10-4
lo-"
10-10
Household vinegar
i
C
b
5 IO-~ 1o - ~ Interior of
lysosomes
L> 6 I 0-8 Cytoplasm o f
--'-&-- working muscle
Neutrality 7 10-7 Pure water at 25°C
8 Seawater
Io -~
@ b lncreaslng
9
10
IO-~
lo-'' Alkallne lakes
alkallnlty 11 10-~~ 0.1 M HCI glass
bx< Household
arnrnonla
y* 12 10-l2 Saturated lime
Figure 3-12 An electrode w~tha proton-seled~vet ~ 1ps a convenient de-
vice for measuring the pH of solutions The t ~ of
p a pH electrode contalns
solutlon a solut~onof pH 7 (I e , [H+] = lo-' M) When the t ~ IS
p Immersed In a so-
13 10-l3 10-1 lut~onof different [H+], the potent~ald~fferenceset up across the enve-
lope of proton-seled~veglass IS propoltlonal to the log of the ratlo of H+
concentrations on the two s~desof the glass
48 PRINCIPLES O F P H Y S I O L O G Y
.........................................
acids, however, have additional carboxyl or amino side
groups that can become acidic or basic. Dissociable side
groups in a macromolecule will determine to a large extent
the electrical properties of the molecule and will addition-
ally render it sensitive to the pH of its environment. This
sensitivity is most dramatically evident in the influence of
pH on the properties of an enzyme's active site. Since the
binding of a substrate to the active site of an enzyme gen-
erally includes electrostatic interactions, the formation of
the enzyme-substratecomplex is highly pH dependent. The
highest probability of binding occurs at a particular pH, the
optimal pH.
Henderson-Hasselbalch Equation
Some acids, such as HCI, dissociate completely, whereas
others, such as acetic acid, dissociate only partially. The
The Biological Importance of pH generalized chemical equation for the dissociation of an
The concentrations of H + and OH- ions are important in acid can be written as
biological systems because protons freely move from H 3 0 +
to associate with and thereby neutralize negatively charged
groups, and OH- ions are available to neutralize positively
charged groups. This ability to neutralize is especially im- in which A- is the anion of the acid HA. Accordingly, the
portant in amino acids and proteins, which are amphoteric dissociation constant derived from the law of mass action
molecules containing both carboxyl (i.e., -COOH) and is given by
amino (i.e. -NH,) groups.
In solution, amino acids normally exist in a dipolar
configuration called a zwitterion:
pH = pK' [A-I
+ log -
[HA1
In other words,
The only amphoteric groups in some amino acids are
the -COOH and -NH, attached to the alpha-carbon atom [proton acceptor]
pH = pK' + log [proton donor]
(C,); these groups enter into peptide bonds. Other amino
MOLECULES, ENERGY, A N D BIOSYNTHESIS 49
......................................
Equation 3-5 is the Henderson-Hasselbalch equation,
which permits the calculation of the pH of a conjugate
acid-base pair, given the pK' and the molar ratio of the pair.
Conversely, it permits the calculation of the pK' ,given the
pH of a solution of known molar ratio.
Buffer Systems
Changes in pH affect the ionization of basic and acidic
groups in enzymes and other biological molecules. Conse-
quently, the pH of intra- and extracellular fluids must be
held within the narrow limits in which enzyme systems
have evolved if these enzymes are to carry out their normal - 0
functions. Deviations of one pH unit or more generally dis- 0.5 1.O
Equivalents of OH-
rupt the biochemistry of organisms. This sensitivity to the
pH of the aqueous intracellular environment exists in part Figure 3-13 The greatest buffering capacity of a conjugate acid-base
because reaction rates of different enzyme systems become system is obtained when pH = pK'. On the graph, this point corresponds
mismatched and uncoordinated. Maintaining the pH of to the part of the curve with the shallowest slope (small pH changes with
large amounts of O H added).
blood is a major goal of the body's homeostatic mecha-
nisms, because large changes in blood pH can be rapidly
transmitted to other body fluids including intracellular
fluids. far greater than that of oils or other nonpolar liquids. The
The pH of body fluids is maintained within normal conductivity of water depends entirely on the presence of
ranges with the help of natural pH buffers. A buffered sys- charged atoms or molecules (ions)in solution. Electrons,
tem is one that tolerates the addition of relatively large which carry electric current in metals and semiconductors,
amounts of an acid or a base with little change in pH over play no direct role in the flow of electric current in aqueous
a certain pH range. A buffer must contain an acid (HA)to solutions. Because the concentrations of H+ and OH-, the
neutralize added bases and a base (Ap)to neutralize added ions present in pure water, are quite low (lo-' M at 2S°C),
acids. (We have already seen that HA is an acid because it the electrical conductivity of pure water is relatively low,
acts as an H+ donor and that A- is a base because it acts though far higher than that of nonpolar liquids. The con-
as an H+ acceptor.) The properties of buffered systems are ductivity of water is greatly enhanced by the addition of
determined by adding small amounts of an acid or base, electrolytes, which dissociate into cations (positive ions)
and recording the pH after each addition. A plot of pH ver- and anions (negative ions) in water (seeFigure 3-7B). Thus,
sus the amount of added acid or base is a titration curve. seawater conducts electrlc current far more readily than
The greatest buffering capacity of a conjugate acid-base freshwater. Spotlight 3-1 reviews some common terms,
pair occurs when [HA] and [Ap] are both large and equal. units, and conventions that apply to electrical properties.
Referring to the Equation 3-5, we see that this situation ex- The role of ions in conducting electric current in solu-
ists when pH = pK' (since log,,, 1 = 0). This point corre- tion is illustrated in Figure 3-14. In this example, two elec-
sponds to that portion of a titration curve along which trodes are immersed in a solution of KC1 and connected by
there is the smallest change in pH (Figure 3-13). wires to a source of electromotive force (emf), the two
The most effective buffer systems are combinations of
weak acids and their salts. Weak acids dissociate only
slightly, thus ensuring a large reservoir of HA. The salts of
weak acids dissociate completely, providing a large reser-
voir of A-. Added H + therefore combines with A- to form
HA, and added O H combines with H+ to form H,O. As
H+ is thereby removed, it is replaced by dissociation of HA.
The most important inorganic buffer systems in the body
fluids are the bicarbonates and phosphates. Amino acids,
peptides, and proteins, because of their weak-acid side
groups, form an important class of organic buffers in the
cytoplasm and extracellular plasma.
- -
106.3 cm long. R = resistlvlty x lengthlcross-sect~onalarea.
. " * 411c7
Resistance Capacitance Battery Variable
resistance
+ -D-
Symbols
4- I-
Ground Switch Meter . Amplifier
terminals marked + and -. The emf causes a current (i.e., rnarily by K+ and C1-; because the concentrations OH-,
a unidirectional displacement of positive electric charge) to H,O+, and H+are so low, their contribution to the current
flow through the electrolyte solution from one electrode to will be ignored. When a potential difference (voltage)is ap-
the other. What does this electric current consist of? In the plied to an electrolyte solution, the cations migrate toward
wire, it consists of the displacement of electrons from the the cathode (electrodewith the negative potential) and the
outer shell of one metal atom to another, then to another, anions migrate toward the anode (electrode with positive
and so on. In the KC1 solution, electric charge is carried pri- potential).
MOLECULES, ENERGY, A N D BlOSYNTHESlS 51
....................................
The rate at which each species of ion migrates in solu-
tion is termed its electrical mobility. This mobility is deter-
mined by the ion's hydrated mass and the amount of charge
(monovalent, divalent, or trivalent) that it bears. The mo-
bility of Ht is considerably higher than the mobilities of
other common ions. The movement of ions that constitutes
an ionic current is crudely analogous to a wave of falling
dominoes, in which each domino (ion) is displaced just
enough to cause a displacement of the next domino. In-
stead of interacting mechanically, like falling dominoes,
ions influence each other through electrostatic interactions,
with like charges repelling each other.
The current in a solution is said by convention to flow
in the direction of cation migration. Anions flow in the Ammeter
opposite direction. The rate at which positive charges are (charge . s-')
displaced past a given point in the solution, plus the rate
at which negative charges are displaced in the opposite di-
rection, determines the intensity of the electric current, Pressure
d~fference
that is, the number of unit charges flowing past a point in a
1second. Thus electric current is analogous to the volume
of water that flows in a second past a point in a pipe
(Figure 3-15). \ /
An electric current always meets some electrical resis-
tance to its flow, just as water meets a mechanical resistance
>-
fr
owing to such factors as friction during its flow through a
pipe. In order for the charges to flow through an electrical
resistance, there must be an electrostaticforce acting on the
charges. This force (analogous to hydrostatic pressure in a
(x) pump Pipe
Anod Carbonyl
ions are based on the same principles that determine ion ex-
change at sites on such nonbiological materials as soil par- Silicate
ticles, glass, and certain plastics. Interactions between k e d
ion-binding sites and various ions are highly important in
certain physiological mechanisms, such as enzyme activa-
tion and the selectivity of membrane channels and carriers Ether
for particular ions.
The energetic basis for interaction between an ion and
an ion-binding site is the electrostatic attraction between Figure 3-18 Many biolog~calmolecules contain groups that exhibit a
partial charge separation. Most common are oxygen-containing groups
the two and is identical in principle to the interactions that
in which the highly electronegative oxygen atom draws electrons from
occur between anions and cations in free solution. Thus, a neighboring atoms. The electron-cloud distributions of several molecu-
site with a nega~ivecharge or a partial negative charge (re- lar slde groups are indicated by shading. Although not present in animals,
call the partial charge on the oxygen atom of the water silicate is a major component of the skeleton of diatoms.
molecule) attracts cations; a site with a positive charge at-
tracts anions. Two or more species of cations in solution * Equation 3-6 should not be confused with Coulomb's law, which is given .
will compete with each other to bind electrostatically to an in the footnote on page 43.
MOLECULES, ENERGY, A N D BlOSYNTHESlS 53
......................................
more rapidly with distance (i.e., a in Equation 3-6 is greater Having discussed some of the basics of the interactions
than 1).This plays an important role in the electrostatic of atoms, elements, and molecules, let us now turn our
tug-of-war experienced by an ion dissolved in water at- attention to those molecules of specific importance to
tracted to a site of opposite charge. organisms.
In an aqueous environment (i.e., in a solution as op-
posed to a vacuum), the coulombic relation (Equation 3-6)
BIOLOGICAL MOLECULES
between the atomic radius of a cation and its affinity for a
given fixed electronegative site is modified by the electro- Even a "simple" unicellular organism has an almost inde-
static interaction of the cation with dipolar water mole- scribably complex molecular makeup. This complexity is
cules. The cation is attracted to both the electron-rich oxy- further compounded by the fact that no two animal species
gen atom of the fixed monopolar site and the electron-rich have the same molecular composition. In fact, the molecu-
oxygen atom of the dipolar water molecule. Thus water lar composition of no individual of a species is identical to
and the site engage in competition for binding of the cation. that of any other in the same species except in those repro-
The more successfully the site competes with water for a duced by cellular fission (e.g., the two daughter cells of an
given ionic species, the greater the "selectivity" of the site amoeba or monozygotous mammalian twins). Such bio-
for that ionic species (Figure3-19). The selectivity sequence chemical diversity is a major factor in evolution, for it pro-
of a site for a group of different ions will be determined by vides an enormous number of variables in a population of
the field strength and the polar/multipolar distribution of organisms and acts as the raw material, so to speak, on
electrons near the site. In addition, the nucleus of a small which natural selection operates. This diversity is in part
atom can more closely approach another atom than can the made possible by the great potential for structural variabil-
nucleus of a large atom. Thus, small monovalent cations ity exhibited by the carbon atom, with its ability to form
will interact more strongly with a particular electronegative four highly stable bonds. In fact, carbon is the "backbone"
site than will large monovalent cations because they carry molecule for the four major classes of organic compounds
the same unit charge but have a smaller distance of closest found in living organisms: lipids, carbohydrates, proteins,
approach. and nucleic acids. We will review the chemical structures of
In addition to the principles of electrostatic interaction these four classes of substances and consider some proper-
briefly described here, there are steric constraints on the ties important to their roles in physiology. More specialized
binding of ions with some sites. If, for example, a site is sit- texts on biochemistry should be consulted for further de-
uated so that an interacting ion must squeeze into a narrow tails (see Suggested Readings).
depression or hollow in or between molecules, the hydrated
size of the ion will then also have an effect on the total en- Lipids
ergy required to reach and interact with the site. Lipids comprise a diverse group of water-insoluble biolog-
ical molecules with relatively simple chemical structures.
The various lipids have a variety of functions. For example,
fats serve as energy stores, while phospholipids and sterols
are major components of membranes (see Chapter 4).
Fats are composed of triglyceride molecules, each of
lnteract~on which consists of a glycerol molecule connected through es-
w~thwater ter bonds with three fatty acid chains. When triglycerides
are hydrolyzed (i.e., digested) by the insertion of H+ and
OH- into the ester bonds, they break down into glycerol
and three fatty acid molecules (Figure3-20). The three fatty
acids in a triglyceride may or may not be the same, but they
all contain an even number of carbon atoms. If all the car-
bon atoms in a fatty acid chain are linked by single bonds
Interaction (i.e., each carbon except the carboxyl carbon bears two hy-
with water drogens), the fatty ac[d is said to be saturated. If the fatty
acid chain contains one or more double bonds between car-
bon atoms, the fatty acid is said to be unsaturated. The de-
gree of saturation and the length of the fatty acids (i.e.,
Figure.3-19 The ability of a fixed anionic site to compete with water mol- number of carbon atoms) composing a fat determine its
ecules for a cation depends on the ion-attracting strength of the site as physical properties.
well as the slze of the cation, because smallercations allow a smaller dis- Fats containing unsaturated fatty acids generally have
tance of closest approach. (A) The force of attraction of a small monova-
low melting- -points and form oils or soft fats at room tem-
lent cation to a strong anionic point site is greater than its attraction t o
water (and vice versa for a large monovalent cation). (B) The force of at-
perature, whereas fats containing saturated fatty acids form
traction of a small monovalent cation t o a weak point site is less than its solids at room temperatures (Table 3-4). That is why the
attraction to water (and vice versa for a large monovalent cation). process of hydrogenation (saturating the fatty acid chains
54 PRINCIPLES OF PHYSIOLOGY
.............................................................................
Figure 3-20 Fats are composed of triglyceride mole-
cules, which are hydrolyzedto glycerol and fatty acids.
This reaction is catalyzed by the enzyme lipase. R rep-
resents a fatty acid radical. The fatty acid groups in a
H-+--o-c~&
&!?- H-t-OH;KCOOH GOOH
+"ii;COOH
+ +
pamcular trlglycerlde may be the same or different.
Fatty acids
H
a Triglyceride Glycerol
with hydrogens and thereby breaking the double bonds) This property allows phospholipid molecules in biological
converts vegetable oil into Crisco, for example. In addition, membranes to form a transition layer between an aqueous
if the number of double bonds is constant, the shorter the phase and a lipid phase. As discussed in the next chapter,
chain length of a fatty acid, the lower its melting point, as biological membranes consist largely of two phospholipid
illustrated by the saturated fatty acids in Table 3-4. Satu- layers, with the nonpolar "tails" of each layer oriented in-
rated fatty acids are more readily converted by metabolic ward toward each other and all the polar "heads" oriented
processes into sterols such as cholesterol. Because excess toward the aqueous phases (see Figure 4-6).
cholesterol appears to be a risk factor for cardiovascular Other types of lipids found in membranes are glyco-
disease in humans, many dietary guidelines recommend lipids, which contain one or more sugar groups, and sphin-
limiting consumption of saturated fats. Cholesterol, how- golipids, which contain a long-chain amino alcohol called
ever, is a component of biological membranes and also is sphingosine. Sphingolipids are present in particularly high
the precursor for synthesis of the steroid hormones (see concentrations in brain and nerve tissue. Waxes constitute
Figure 9-23). another group of lipid; they form an important water-
Triglycerides typically accumulate in the fat vacuoles of proofing layer in certain insects (see Chapter 14).
specialized adipose cells in vertebrates. Because of their low
solubility in water, these energy-rich molecules can be Carbohydrates
stored in large concentrations in the body without requir- Carbohydrates are polyhydroxyl aldehydes and ketones
ing large quantities of water as a solvent. Triglyceride en- with the general chemical formula of (CH,O), . The sim-
ergy stores are also rendered highly compact by the rela- plest carbohydrates are the monosaccharide sugars, the
tively high proportions of hydrogen and carbon and low most common of which contain six carbons (hexoses) or
proportions of oxygen in the molecule. Thus, 1g of triglyc- five carbons (pentoses).Monosaccharides typically exist
eride will yield about two times as much energy upon oxi- as ring structures containing four or five carbon atoms
dation as 1g of carbohydrate (Table 3-5). and one oxygen, with the remaining carbon(s) outside the
In phospholipids one of the outer fatty acid chains of ring (Figure3-21A). Green plants manufacture the hexose
a triglyceride is replaced with a phosphate-containing glucose from H,O and CO, by the process of photosyn-
group (see Figure 4-3). Thus the phospholipids are amphi- thesis. All the energy trapped by photosynthesis and trans-
pathic molecules, with a hydrophilic portion (the phos- mitted as chemical energy to the living world (i.e., all plant
phate-containing group) and a hydrophobic portion (the and animal tissues) is channeled through such six-carbon
fatty acid chains), which is soluble in lipids (or lipophilic). sugars as glucose. As noted later in this next chapter, the
complete or partial degradation of glucose to H,O and
C 0 2 during cellular respiration releases the chemical en-
TABLE 3-4 ergy that was stored in its molecular structure during pho-
Melting points of various fatty acids tosynthesis. The two most important pentose sugars are
No. of No. of Melt~ng ribose and 2-deoxyribose (see Figure 3-21A). These pen-
carbon double polnt toses, which occur in the backbones of all nucleic acid mol-
Fatty ac~ds atoms bonds PC) ecules, are essential for replication of DNA and synthesis of
Saturated proteins.
Lauric acid 11 0 44
Palm~ticacid 16 0 63
TABLE 3-5
Arachidlc acid 20 0 75 The energy content of the three major categories of foodstuffs
Llgnoceric acid 24 0 84
Energy content
Unsaturated Substrate (kcal .g-')
Proteins
Proteins are the most complex and the most abundant or-
ganic molecules in the living cell, making up more than half
the mass of a cell as measured by dry weight. Although the
basic structure of all proteins is similar, a vast array of dif-
ferent proteins with diverse functions is found in biological
Lactose
systems. Table 3-6 lists the major functional types of pro-
teins with several examples of each type. Enzymes consti-
Figure 3-21 The simple sugars are monosaccharides and disaccharides. tute the largest functional group of proteins, with more
(A) Glucose, the most prevalent hexose in cells, is degraded t o provide than 1000 already identified and many unknown ones pre-
energy. The hydroxyl groups in red can form a covalent bond with an-
sumably to be discovered.
other sugar molecule, forming a disaccharide. Two pentoses, ribose and
2-deoxyribose, are constituents of nucleic acids. (B) Disaccharides are
formed by condensation of two monosaccharideunits. Sucrose and lac-
Primary structure
tose both contain one glucose unit (shaded) plus a second monosac-
charide. The glycosidic bond (red) linking two monosaccharideunits can
Proteins are composed of linear chains of amino acids,
have two different orientations, designated a and P. which are amphoteric molecules containing at least one
carboxyl group and one amino group. The 20 common
amino acids that make up proteins are all alpha-amino
Cells contain enzymes that can convert glucose to other acids, in which the amino group is bonded to the alpha-
monosaccharides or link two monosaccharide molecules carbon (C,) atom, that is, the carbon atom adjacent to the
to form a disaccharide sugar such as sucrose or lactose carboxyl group. Amino acids differ from one another in the
(Figure 3-21B). Cells also can synthesize various carbohy- structure of their side groups, generically referred to as R
(1 - 4) chain
Figure 3-22 Glycogen, a large glucose polymer, IS the prlmary carbo- I~nked,w~thbranches extending from carbon 6 every elght to ten glucose
hydrate storage form In anlmal cells Aglycogen molecule IS a long cham resldues Only a small port~onof a glycogen molecule IS dep~cted.
of glucose resldues, In whlch carbons 1 and 4 in adjacent molecules are
56 PRINCIPLES OF PHYSIOLOGY
...............................
Figure 3-23 Chitin, a structural carbohydrate
units joined by 1-
polymer, consists of N-acetyl glucosamine
4 glycosidic bonds. In
N-acetyl glucosamine, an acetamide group
(colorshading) replaces the hydroxyl group on
carbon 2 of glucose.
groups (Figure 3-24A). The protein-synthesizing machin- rial. Indeed, all the hereditary information carried in the
ery of cells joins amino acid molecules via covalent peptide genetic material is translated initially into protein mole-
bonds, forming long polypeptide chains. Adjacent Ca cules: the amino acid sequence laid down during protein
atoms in a polypeptide chain are separated by a planar synthesis is the expression of this information and is the
amide group (Figure 3-24B).The specific linear sequence of primary determinant of the properties of any protein mol-
amino acid residues of a polypeptide is termed its primary ecule. Since there are about 20 different amino acid build-
structure. Since the amino acid residues of a polypeptide ing blocks, an impressive variety of different amino acid
chain differ only in their side groups, these groups are like sequences is possible. Suppose, for example, that we were
letters in the protein alphabet, defining the primary struc- to construct a polypeptide molecule consisting of one of
ture of a protein (Table 3-7). A protein molecule may con- each of those 20 building blocks. How many different lin-
sist of one, two, or several polypeptide chains, either cova- ear arrangements could we make without ever repeating
lently linked or held together by weaker bonding. the same sequence of amino acids? This is determined by
The amino acid sequence of a polypeptide (i.e., its pri- multiplying20X 1 9 X 18 X 1 7 X 1 6 X . . . X 2 X 1
mary structure) is encoded in an organism's genetic mate- (i.e., 20!), or 1018. But this startlingly large figure, which
TABLE 3-6
Classification of proteins and peptides according to biological function
Type/examples Occurrence or function Type/examples Occurrence or function
Amide
group
Peptide
bond
-CH,-CH2-C
3.6 amino acid residues per turn. The side group of each amino acid proline, for example, the side group is a rigid
amino acid residue extends outward from the helical back- ring that includes the alpha-nitrogen atom (see Table 3-7).
bone, free for interaction with other side groups or other Thus, the C6N bond in proline (and hydroxyproline)can-
molecules. The stability of the a helix is enhanced substan- not rotate; as a result, whenever proline (or hydroxypro-
tially by hydrogen bonding between the oxygen atom of a line) occurs in a peptide chain, it interrupts the a helix,
carbonyl group and the hydrogen atom of the amide group causing the peptide backbone to bend.
four residues ahead. Because of the stability of the a helix, Another major type of protein secondary structure is
a polypeptide chain spontaneously assumes this conforma- the beta (p)pleated sheet (Figure3-26). This consists of lat-
tion, provided that the side groups do not interfere. In the erally associated P strands, which are fairly short, nearly
MOLECULES, E.NERGY, A N D BIOSYNTHESIS 59
. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. . . . . . . . . . . . . . . . . . . . . . . . .
Face view Figure 3-26 The P pleated sheet is an ele-
ment of secondary structure in silk fibers and
some other fibrous proteins. Pleated sheets
are formed by the lateral association of two b
strands stabilized by hydrogen bonds (black
dots). The side groups (R) extend above and
below the plane of the sheet. The broad ar-
rows represent b strands. [Adapted from
Lodish et al., 1995.1
Side view
fully extended stretches of the polypeptide chain. Hydro- tor to the conformation of proteins is the sulfhydryl side
gen bonding between carbonyl oxygen atoms and amide group (-SH) of the amino acid cysteine. The reaction of
hydrogen atoms in adjacent P strands forms a pleated sheet two cysteine residues forms a disulfide linkage (S-S), which
with the side groups of the amino acid residues projecting covalently joins the residues (Figure 3-27).A disulfide link-
above or below the plane of the sheet. Association of P age can covalently cross-link different portions of a poly-
strands within the same polypeptide chain contributes to peptide chain, thereby stabilizing its folded tertiary struc-
secondary structure, whereas association of p strands in ture, or connect two separate chains. Since the sulfhydryl
different polypeptide chains contributes to quaternary group is highly reactive, it is not surprising that one or more
structure. cysteine residues frequently occupy the active sites of en-
Long polypeptide chains with an uninterrupted a-helix zymes. The toxicity of mercury and other heavy metals is
conformation are characteristicof fibrous proteins, such as due, in part, to their reaction with the sulfur atom of cys-
the alpha-keratins that form hair, fingernails and claws, teine, displacing the hydrogen atoms. This reaction can poi-
wool, horn, and feathers. Beta-keratins are an exception, son (i.e., render catalytically inoperative) the active site of
having a secondary structure consisting of P pleated sheets an enzyme.
' rather than a helices. Beta-keratins are the major con- Some, but not all, proteins undergo self-assembly,
stituent of spider webs and silk, which is produced by cater- forming a quaternary structure. The amino acid sequence
pillars. Nonstructural intracellular proteins typically have of a polypeptide chain-and hence the positions of the
a random-coil secondary structure, although these proteins different amino acid side groups-not only determines
may contain short segments in the a-helix or P-pleated- the secondary and tertiary structure of the molecule,
sheet conformation. but also may allow interaction with other polypeptide
Regions of a polypeptide chain with long a helices of- chains, thereby forming protein molecules with two
ten assume a rodlike tertiary structure, whereas those lack- or more subunits. The association of subunits can involve
ing this feature have a globular tertiary structure. Two types covalent disulfide linkages between them as well as nonco-
of relatively weak, noncovalent interactions help stabilize valent interactions between complementary regions on
tertiary structure: coulombic (electrostatic)interactions be- their surfaces. For example, negatively charged groups
tween the charged side groups and van der Waals forces be- of one subunit fit against positively charged groups of an-
tween hydrophobic side groups. Another major contribu- other subunit; hydrophobic, nonpolar side groups on the
H
I
2 @ r ~ ~ , - ~ - Cysteine
~ ~ ~ ~
I
N"2
I
Cystine
~isulfide
linkage Nucleic Acids
Figure 3-27 A disulfide bond can contribute to the tertiary structure of Deoxyribonucleic acid (DNA)was first isolated from white
proteins by linking cyteine residues present in different portions of the blood cells and fish sperm in 1869 by Friedrich Miescher.
same polypeptide chain. Disulfide bonds also can form between cysteine ~~~i~~the next decadesthe chemical composition of DNA
residues in different polypeptide chains, thereby contributing to quater-
nary structure.
was gradually worked out, and evidence slowly accumu-
lated that implicated it in the mechanisms of heredity. We
now know that DNA, which is associated with the chro-
mosomes, carries coded information, arranged into genes,
subunits meet to the mutual exclusion of water molecules;
that is passed from each cell to its daughter cells and from
or residues in each subunit are oriented so they can form
one generation of organisms to the next. A second group of
hydrogen bonds. Some enzymes, the respiratory pigment
nucleic acids, ribonucleic acid (RNA),was subsequently
hemoglobin, and many other proteins consist of more than
discovered. RNA is now known to be instrumental in
one polypeptide chain held together by noncovalent bonds.
translating the coded message of DNA into sequences of
In some multi-subunit proteins, /Ipleated sheets connect
amino acids during synthesis of protein molecules.
the subunits. The three subunits of collagen, the major pro-
The nucleic acids are polymers of nucleotides, which
tein in connective tissue, are twisted into a characteristic
consist of a pyrimidine or purine base, a pentose sugar, and
superhelix (Figure 3-28). The subunits of all these pro-
a phosphoric acid residue (Figure 3-29). The nucleotides
teins will assemble themselves spontaneously if added sep-
composing DNA contain deoxyribose, whereas those com-
arately to an aqueous solution and mixed. The associated
posing RNA contain ribose (see Figure 3-21A).The major
and dissociated subunits of hemoglobin are illustrated in
nucleotides found in nucleic acids contain the following
Figure 13-2A.
bases: adenine, thymine, guanine, cytosine, and uracil.
Except for covalent disulfide linkages between cysteine
Thymine occurs only in DNA, and uracil only in RNA; the
residues, the secondary, tertiary, and quaternary structure
other three bases are found in both nucleic acids. Stable
of proteins depends on coulombic interactions, hydrogen
base pairs, linked by hydrogen bonds, can form between
bonding, and van der Waals forces. All of these noncova-
adenine and thymine (A-T), guanine and cytosine (G-C),
lent interactions are relatively weak and heat labile. Heat-
and adenine and uracil (A-U), as depicted in Figure 3-30.
ing a protein disrupts these interactions leading to alter-
In a polynucleotide chain, phosphodiester bonds link the 3'
ations in its conformation, called denaturation. Hair
carbon of one pentose ring and the 5' carbon of the next
curling irons work in this way, temporarily heating the pro-
pentose (Figure 3-31). The purine and pyrimidine bases ex-
teins in the hair shaft and then letting them cool in slightly
tend outward from the polynucleotide backbone and are
new configurationsthat alter the shaft's orientation. In this
not involved in the repetitive, nonvarying backbone.
same way, high temperatures can change the shape of en-
Native DNA consists of two chains (or strands) in
zymes, rendering them inactive and killing the cells in
which the sequence of bases is complementary (e.g., an ade-
which they reside.
d
5' structure conslstlng of a purine or pyrlml-
$- d~nebase, a pentose sugar, and a phos-
phoric ac~dres~dueP In DNA,the pen-
tose IS 2-deoxyribose, wh~chhas two
H 3'
hydrogen atoms attached to the C, atom,
In RNA, one of these hydrogens, lnd~cated 5r 0 H
OH W by shading, IS replaced by a hydroxyl 1
group. HO-P=O
I *h@:
0 I
deoxyribose
0 H
I
Figure 3-31 The backbone structure of polynucleotide cha~nsconslsts
-
GUANINE CYTOSINE BASE PAIR of pentose res~dueslinked by phosphod~esterbonds The bases extend
away from the backbone Th~sdiagram shows a small portion of a single
Guanine Cytosine strand of DNA. [Lehninger, 19751
6h
b
The genetic information of an organism is encoded
in the sequence of bases in its DNA. In a process called
transcription, a DNA strand acts as a template for syn-
thesis of messenger RNA (mRNA) in the nucleus (see
Figure 3-32, bottom). The mRNA strand, which contains
the informational sequence present in its DNA template,
leaves the nucleus and enters the cytoplasm to be decoded
deoxyribose
C-1' of '
deoxyribose
by a ribosome into the amino acid sequence of a polypep-
tide chain. In this process of translation, certain sequences
of three bases in the DNA code for certain amino acids. For
Figure 3-30 In nucleic acids, hydrogen bonding (black dots) between
purine and pyrimidine bases forms the stable base pairs G-C, A-T (in
example, GGU, GGC, CGA, and GGC all code for the
DNA),and A-U(in RNA).The structure of uracil (U) is the same as thymine amino acid glycine; and GCU, GCC, GCA, and GCT code
except that the methyl (-CH,) group on C, is replaced with a hydrogen. for the amino acid alanine. Thus, the genetic code consists
62 PRINCIPLES O F P H Y S I O L O G Y
.........................................
DNA DNA ENERGETICS OF LIVING CELLS
The biochemical reactions of animal cells are in many
senses those of a chemical machine. As in all machines,
each event is accompanied by an energy transaction. For a
machine or a cell to perform work, energy must be trans-
ferred from one part of the system to another, usually with
the conversion of at least part of the energy from one form
to another. This holds true even when the parts of the sys-
tem are as minute as reacting molecules.
Animals are fueled by the intake of organic food mol-
ecules and their subsequent degradation by digestive and
metabolic processes. During these processes the chemical
energy inherent in the molecular structures of foods is re-
leased and made available for the energetic needs of the or-
ganism. Energy is required for such obvious activity as
muscle contraction, ciliary movement, and the active trans-
port of molecules by membranes. However, chemical en-
ergy is also required for the synthesis of complex biological
molecules from simple chemical building blocks and for the
subsequent organization of these molecules into organelles,
cells, tissues, organ systems, and complete organisms. A liv-
ing organism must frequently take in fuel and continuously
expend energy to maintain its function and structure at all
levels of organization. If energy intake drops below the
transcribed amount required for maintenance, the organism will con-
sume its own energy stores. When these are exhausted, it
no longer has any source of energy. The organism dies be-
cause it cannot stave off the tendency to become disorga-
nized, nor can it continue to perform the necessary energy-
requiring functions.
The material and energy transactions that take place in
an organism constitute its metabolism. At the intracellular
level, these transactions take place via intricate reaction se-
quences called metabolic pathways, which in a single cell
can involve thousands of different kinds of reactions. These
DNA mRNA DNA reactions do not occur randomly, but in orderly sequences,
regulated by a variety of genetic and chemical control
Figure 3-32 Native DNAcontainstwostrands wound around each other
mechanisms. The organization of atoms and molecules
in a double helix. Hydrogen bonding (black dots) between cornplemen-
tary bases stabilizes the structure. The molecule unwinds during tran- into highly specific structures plus the ability to carry out
scription, and one of the strands acts as a template for synthesis of mRNA cellular metabolism distinguishes living systems from the
complementary to the DNA. nonliving.
The processes of cell metabolism in animals are of two
kinds:
of a four-letter alphabet (A, G, C, T) combined into three-
letter words. Extraction of chemical energy from foodstuff molecules
As discussed in the previous section, the primary struc- and the channeling of that energy into useful functions.
ture (amino acid sequence) of a polypeptide determines its Chemical alteration and rearrangement of nutrient
final three-dimensional conformation. Thus once a poly- molecules into small precursors of other kinds of bio-
peptide chain is synthesized, it curls and folds, assuming the logical molecules.
characteristic secondary and tertiary structure of a protein
molecule, and in some cases associates with other chains to An example of extraction is the acquisition of amino acids
form a multi-subunit protein. during the digestion of foodstuff proteins and their subse-
More detailed accounts of the major steps relating pro- quent oxidation within cells, which releases their chemical
tein synthesis to nucleic acids may be found in the refer- energy. An example of alteration and arrangement is the in-
ences listed under Suggested Readings at the end of the corporation of amino acids into newly synthesized protein
chapter. molecules in accord with the specifications of the genetic
information of the cell. We are concerned here less with or coal to fuel a steam engine, this does not create new en-
the biochemical details of cell metabolism than with the ergy, but merely converts one form to another-in this ex-
thermodynamic and chemical principles that underlie the ample, chemical energy to thermal energy, thermal energy
transfer and utilization of chemical energy within the cell. to mechanical energy, and mechanical energy to work.
Thus, we will consider the mechanisms by which chemical The second law of thermodynamics states that all the
energy is extracted from foodstuff molecules and the man- energy of the Universe will inevitably be degraded to heat
ner in which it is made available for the energy-requiring and that the organization of matter will become totally ran-
processes discussed in subsequent chapters. domized. In more formal terms, the second law states that
the entropy, a measure of the randomness, of a closed sys-
Energy: Concepts and Definitions tem will progressively increase and that the amount of
Energy may be defined as the capacity to do work. Work, in energy within the system capable of performing useful
turn may be defined as the product of force times distance work will diminish. A system that is ordered (nonrandom)
( W = F x d ).As an example, when a force lifts a 1-kg mass contains energy in the form of its orderliness, because
a height of 1m, the force is 1kg, and the mechanical work in becoming disordered (i.e., as a result of an increase in
done is 1m - kg. The energy expended to do this work (i.e., entropy), it can perform work. This is illustrated
the useful energy, not including that expended in overcoming in Figure 3-33A, which shows gas molecules in thermal
friction or expended as heat) is also 1m .kg. Once the kilo- motion in a hypothetical system consisting of two com-
gram mass is raised to the height of 1 m, it possesses, by partments open to each other. Initially the gas is confined
virtue of its position, a potential energy of 1m .kg. This po- almost entirely to compartment I, in which case the system
tential energy can be converted to kinetic energy (energy of possesses a certain degree of order. Clearly, this situation
movement) if the mass is allowed to drop. Thus, we see that has a very small probability of occurring spontaneously if
energy exists in different forms, including the following: in the starting condition the gas molecules are evenly dis-
tributed between the two compartments. The gas molecules
Mechanical potential energy (e.g., a stretched spring or can all be forced into one compartment only by the expen-
a lifted weight) diture of energy (e.g., a piston pushing the gas from one
Chemical potential energy (e.g., gasoline, glucose) compartment to the other). As the gas is permitted to es-
cape from compartment I into compartment 11, the entropy
Mechanical kinetic energy (e.g., a falling weight) of the system increases (i.e., the system becomes more ran-
Thermal energy (actually kinetic energy at the molecu- dom). The movement of molecules from compartment I to
lar level) compartment 11is a form of useful energy that can be made
Electrical energy to do work on an appropriate apparatus placed near the
opening between the two compartments. Once the system
0 Radiant energy is fully randomized (i.e., entropy is maximal), no further
work can be extracted from the system, even though
The various forms of energy can power different types the gas molecules remain in constant thermal motion
of work, as summarized in Table 3-8. We will be concerned (Figure 3-33B).
in this chapter primarily with chemical energy, the poten- Orderliness increases as the organism develops from a
tial energy stored in the structure of molecules. Before delv- fertilized egg to the adult. In this sense, living systems tem-
ing into the energy relationships involved in the biochemi- porarily defy the second law. It should be recalled, however,
cal reactions of cellular metabolism, it will be useful to that the second law refers to a closed system (e.g., the Uni-
review the first and second laws of thermodynamics and verse), and animals are not closed systems. Living orga-
the concept of free energy. , nisms maintain a relatively low entropy at the expense of
energy obtained from their environment. For example, a
Thermodynamic laws rhinoceros eating, digesting, and metabolizing grass in
T h e first law of thermodynamics states that energy is nei- quantities just sufficient to maintain constant weight ulti-
ther created nor lost in the Universe. Thus, if we burn wood mately increases the entropy of the matter it ingests. The
highly ordered carbohydrate, protein, and fat molecules in
TABLE 3-8 the grass are converted in the animal to CO,, H,O, and
Various kinds of work low-molecular-weightnitrogen compounds, releasing en-
Displacement ergy trapped in the organization of the larger molecules
Type of work Driving force variable (Figure 3-34). The carbon, hydrogen, and oxygen atoms in
Expansionwork P (pressure) Volume cellulose, for instance, are in a much more highly ordered
Mechanical work F(force) Length state than they are in C 0 2 and H,O; thus the metabolic
Electricalwork €(electric potential) Electric charge
breakdown of cellulose in the grass represents an increase
in entropy. At the same time, the cells of the rhinoceros
Surface work r (surfacetension) Surface area
utilize for their own energy requirements a portion of
Chem~calwork p(chemical potential) Mole numbers
the chemical energy originally stored in the molecular
64 PRINCIPLES OF PHYSIOLOGY
...............................................................................
I\ Figure 3-33 Low- and hlgh-entropystates can be ex-
I
Compartment I plalned by a mechanical analogy In (A) nearly all the
gas molecules are In compartment I, an organized,
hlgh-energy state As the molecules are allowed to
dlffuse Into compartment 11, the entropy of the sys-
Work belng tem Increases and the useful energy decreases untll
ach~eved equlllbrlum IS reached (B). The change from a low- to
a hlgh-entropy state releases useful energy, whlch In
thls model IS harnessed by the paddle wheel The
ab~lltyto do work approaches zero as the system
comes Into equlllbrlum [Adapted from Baker and
Allen, 1965 ]
Compartments in
equilibrium
organization of foodstuff molecules. This is not in conflict organism. For this reason, biological systems can utilize
with the second law because the decrease in entropy that re- only that component of the total available chemical energy
sults from the animal's synthesis of complex molecules oc- capable of doing work under isothermal conditions. This
curs at the expense of increasing the entropy of ingested component is called the free energy, symbolized by the let-
foodstuff molecules produced by plants with the energy of ter G. Changes in free energy are related to changes in heat
the sun. Ultimately, of course, the rhinoceros dies, and the and entropy by the equation
entropy of its body greatly increases as it decays or is con-
sumed by other animals. AG=AH-TAS (3-7)
i
pends on radiant energy from the sun. This electromagnetic
energy (including visible light) has its origin in nuclear fu-
sion, a process in which the energy of atomic structure is
converted to radiant energy. In this process, four hydrogen
nuclei are fused to form one helium nucleus, with the re-
lease of an enormous amount of radiant energy. A very
small fraction of this radiant energy reaches the planet
d'
Paraites I Carnrres
-
pathways by which animal cells release energy through the
oxidation of food molecules. First, however, it will be use- Sometimes, however, the tendency for a reaction to go for-
ful to examine some general principles of energy transfer in ward (reactants products) is so much greater than the
biochemical reactions, and also some features of enzymes, - tendency to go in reverse that for practical purposes the re-
the cellular proteins that allow biochemical reactions to action may be considered irreversible.
proceed rapidly at biological temperatures. A reaction tends to go forward if it shows a free-energy
change, AG, that is negative. In other words, the total free
Transfer of Chemical Energy by Coupled Reactions energy of the reactants exceeds that of the products. Such
There are several categories of biochemical reactions, but reactions are said to be exergonic (or exothermic) and typ-
the features of reaction rates and kinetics can be illustrated ically liberate heat. The oxidation of hydrogen to water is
by a simple combination reaction in which two reactant a simple exergonic reaction:
molecules, A and B, react to form two new product mole-
cules, C and D: 2 H, + 0, - 2 H,O + heat
66 PRINCIPLES OF PHYSIOLOGY
.......................................
The energy-requiring reverse reaction occurs during pho-
tosynthesis, the energy being supplied by chlorophyll-
trapped light quanta:
Here [A], [B], [C], and [Dl are the equilibrium molar con-
centrations of the reactants and products in Equation 3-7.
It is evident that the greater the tendency for the reaction in
Equation 3-7 to go to the right, the higher the value of its
K:,. As noted already, this tendency depends on the differ-
ence in free energy, AG, between the products C and D and
Figure 3-36 In this mechanical analogy of a coupled reaction, the fall of
the reactants A and B. The greater the drop in free energy, the 10-kgweight providesthe energy required to lift the 3-kg weight. The
the more completely the reaction proceeds to the right and pulley and rope connecting the two weights is the mechanism for cou-
the higher its KLq.The equilibrium constant is related to the pling the energy of the falling 10-kg weight t o the other weight.
change in standard free energy, AGO, of the system by the
equation
tential energy (10 m .kg) by dropping a distance of 1m, in
which case it will lift the 3-kg weight on the right the same
distance. Because the two weights are connected with a
It is evident from this equation that if K:, is greater than rope over a pulley, the fall of the 10-kg weight is coupled to
1.0, AGO will be negative; and if K:, is less than 1.0, AGO the rise of the 3-kg weight, which initially had no potential
will be positive. Exergonic reactions have a negative AGO energy of its own. It is evident that the falling weight can
and therefore occur spontaneously without the need of ex- raise the other one only if it weighs more. Likewise, an ex-
ternal energy to "drive" them. Endergonic reactions have a ergonic reaction can "drive" an endergonic reaction only if
positive AGO; that is, they require the input of energy from the former liberates more free energy than the latter re-
a source other than the reactants. quires. As a consequence, some energy is lost, and the effi-
Some biochemical processes in living cells are exergonic ciency is, of necessity, less than 100%.
and others are endergonic. Exergonic processes, since they
proceed on their own under the appropriate conditions, ATP: Energy Carrier of the Cell
present relatively few problems in cellular energetics. En- The most ubiquitous energy-rich common intermediate in
dergonic processes, however, must be "driven." This is gen- cellular metabolism is the nucleotide adenosine triphos-
erally done in the cell by means of coupled reactions, in phate (ATP), which can donate its terminal energy-rich
which common intermediates transfer chemical energy phosphate group to any of a large number of organic ac-
from a molecule of relatively high energy content to a re- ceptor molecules (e.g., sugars, amino acids, nucleotides).
actant of lower energy content. As a result, the reactant This phosphorylation reaction raises the free-energy level
is converted into a molecule of higher energy content and of the acceptor molecule, allowing it to react exergonically
can undergo the required reaction by releasing some of this in enzyme-catalyzed biochemical reactions.
energy. The ATP molecule consists of an adenosine group,
A mechanical analogy of a coupled reaction is seen in made up of the pyrimidine base adenine, the five-carbon
Figure 3-36. The 10-kg weight on the left can lose its po- sugar residue ribose, and three linked phosphate groups
(Figure 3-37A).Much of the free energy of the molecule re- is highly endergonic. The standard free-energy change,
sides in the mutual electrostaticrepulsion of the three phos- AGO, for the hydrolysis of ATP under standard conditions
phate units, with their positively charged phosphorus is -7.3 kcal .mol-'.
atoms and negatively charged oxygen atoms. The mutual The role of ATP in driving otherwise endergonic reac-
repulsion of these phosphate units is analogous to the re- tions by means of coupled reactions is illustrated by the
pulsion of bar magnets, with their north and south poles condensation of the two compounds X and Y to yield Z:
aligned, held together by a sticky wax (Figure 3-38). If the
-
wax, which is analogous to the 0 P bonds in ATP, is soft- X + ATP '---.I X-phosphate + ADP
ened by warming, the energy stored by virtue of the prox-
AGO = -3.0 kcal .mol-I
imity of the mutually repelling magnets is released as the
magnets spring apart. Likewise, the breaking of the bonds X-phosphate +Y Z + P,
between the phosphate units of ATP results in the release of
free energy (Figure 3-37B). Once the terminal phosphate
group of ATP is removed by hydrolysis, the mutual repul-
The total free energy liberated in these two reactions
sion of the two products, adenosine diphosphate (ADP)
(-5.3 kcal.mol-l) will be equal to the sum of the free-
and inorganic phosphate (PI),is such that the probability of
energy changes of the two parent reactions:
their recombining is very low. That is, their recombination
ATP + HOH ADP + Pi
A AGO = -7.3 kcal .mol-l
Adenosine group
X + Y Z
AGO =
+ 2.0 kcal .molpl
- 5.3 kcal mol-I
Wax
I
u
ADP (lower energy form)
A
I
OH
CH3
Glucose 1-phosphate
0-NH-c-LcH,-coo-
t II
+NH2 AGo = y ~ z Q ~ ~ ~ &
Phosphocreatine
AGO =
Acetyl phosphate
AGO =
Glucose 6-phosphate
NH3+
B-NH-C-NH-(CH,),-AH-~~O-
t II
+NH2
Phosphoarginine
Glycerol &phosphate
MOLECULES, ENERGY, A N D BIOSYNTHESIS 69
...............................................................................
Both creatine phosphate and arginine phosphate are found
singly or together in invertebrate muscle. , Activated S without
enzyme; AG* much higher
/%
Temperature and Reaction Rates
The rate at which a chemical reaction proceeds depends on
the temperature. This is not surprising, because tempera- I
/t
t
I
AG*
:,'
ture is an expression of molecular motion. As temperature %
I
I
increases, so does the average molecular velocity. This $
C
AG+ / ES*
greater velocity increases the number of collisions per unit
time and thereby increases the probability of successful in-
teraction of the reactant molecules. Furthermore, as their
:
?! E + S
LL --I
I
\
act is called the free energy of activation, or activation \
r Activation
energy combustion of the H, produces a rapid ignition of the re-
maining unburned H,. In contrast, when H2 is oxidized
slowly at a low temperature with small quantities of the
catalytic agent platinum, the release of heat is slowed
enough so that no explosion occurs. The quantity of plat-
inum relative to the fuel (H,) and oxidant (0,)regulates
the rate of combustion. Likewise, most biological reactions
are regulated by modulation of the quantity or the catalytic
effectiveness of certain enzymes. In the next two sections,
Figure 3-40 The activation energy is the energy required to bring the re-
actants into position t o interact. In this analogy, the potential energy of
we'll first discuss how enzymes operate and then how cells
the rock cannot be liberated until some energy, referred to as the activa- regulate their metabolic reactions by controlling the syn-
tion energy, is expended to bring it into position at the crest of the hill. thesis and catalytic activity of enzymes.
70 PRINCIPLES OF PHYSIOLOGY
.......................................
ENZYMES: GENERAL PROPERTIES
Substances that increased the rate of alcoholic fermenta-
tion were first isolated from living cells by water extrac-
tion of yeast about a century ago. These substances, now
called enzymes, were found to be inactivated by heating,
whereas the reactants in fermentation reactions were un-
affected by heating. This finding was the first indication
that enzymes are protein molecules. It was subsequently
discovered that, without exception, each species of en-
zyme molecule is a protein of very specific amino acid
composition and sequence. All of these proteins, or at least
their enzymatically active portions, have a globular con-
formation. Each cell in an organism contains literally
thousands of species of enzyme molecules catalyzing all
the synthetic and metabolic reactions of the cell. The work
of molecular geneticists has demonstrated that enzymes
are the primary gene products of major significance. By
specifying the structure of each enzyme molecule that is Figure 3-42 Chymotrypsin hydrolyzes any peptide bond in which the
carbonyl carbon belongs to a phenylalanine, tyrosine, or tryptophan
produced, the genetic apparatus is indirectly responsible
residue. Shown here is its action on a dipeptide containing phenylalanine
for all enzymatic reactions in a cell. (shading).
Whatever the precise catalytic mechanism for a par- Figure 3-44 Both temperature and pH influencethe activity of enzymes.
(A)The effect of temperature on the reaction rate is generally similar for
ticular reaction, once the substrate molecules have most enzymes. (B)The effect of pH on catalytic activity varies among en-
reacted, the enzyme separates from the products, free- zymes, but nearly all have a d~st~nct
optimal pH.
ing the enzyme molecule to form an ES complex with a
new substrate molecule. Since the ES persists for a finite
time, all the enzyme can become tied up as ES if the sub- ever, the rate of enzyme inactivation due to denaturation
strate concentration is high enough relative to the enzyme also increases. At the optimal temperature, the rate of en-
concentration. zyme destruction by heat is balanced by the increase in en-
zyme-substrate reactivity, and the two effects of elevated
Effect of Temperature and pH on temperature cancel. At that temperature the reaction rate is
Enzymatic Reactions maximal. At higher temperatures enzyme destruction
Any factor that influences the conformation of an enzyme, becomes dominant, and the rate of reaction rapidly de-
and hence the arrangement of amino acid side groups in the creases. The temperature sensitivity of enzymes and other
active site, will alter the activity of the enzyme. Tempera- protein molecules contributes to the lethal effects of exces-
ture and pH are two common factors that influence the sive temperatures.
rates of enzymatic reactions in this way. Electrostatic bonds often participate in the formation
As we saw previously, an increase in temperature in- of an ES. Since H+ and OH- can act as counterions
creases the probability of protein denaturation, which dis- for electrostatic sites, a drop in pH exposes more positive
rupts the conformation of polypeptide chains. In the case sites on an enzyme for interaction with negative groups on
of enzymes, denaturation destroys catalytic activity. a substrate molecule. Conversely, a rise in pH facilitates
For this reason, enzyme-catalyzed reactions exhibit a the binding of positive groups on a substrate to negative
characteristic curve of reaction rate versus temperature sites on the enzyme. Thus, it is not surprising that the
(Figure 3-44A). As temperature increases, the reaction rate activity of enzymes typically varies with the pH of the
initially increases due to the increased kinetic energy of the medium and that each enzyme has an optimal pH range
substrate molecules. As temperature increases further, how- (Figure 3-44B).
Cofactors in limiting concentrations for certain enzymes. As discussed
Some enzymes require the participation of small molecules in Chapter 9, the Ca2+concentration of the cytosol is reg-
called cofactors in order to perform their catalytic function. ulated by the surface membrane and by internal organelles,
In that case, the protein moiety is called the apoenzyme. One such as the endoplasmic reticulum. In this way, the activ-
class of cofactors consists of small organic molecules called ity of calcium-activated enzymes can be regulated by the
coenzymes, which activate their apoenzymes by accepting cell. Cellular processes regulated by the Ca2+concentration
hydrogen atoms (protons)from the substrate. For example, include muscle contraction, secretion of neurotransmitters
the enzyme glutamate dehydrogenase requires the coenzyme and hormones, ciliary activity, assembly of microtubules,
nicotinamide adenine dinucleotide (NAD)to catalyze the and ameboid movement.
oxidative deamination of the amino acid glutamic acid:
Enzyme Kinetics
glutamate + NAD+ The rate at which an enzymatic reaction proceeds depends
(oxidized) on the concentrations of substrate, product, and active en-
a-ketoglutarate + NADH + NH,+ zyme. For purposes of simplicity, we will assume that the
(reduced) product is removed as fast as it is formed. In that case, the
rate of reaction will be limited by the concentration of ei-
A number of coenzymes contain vitamins as part of the ther the enzyme or the substrate. If we further assume that
molecule. Since an apoenzyme cannot function without its the enzyme is present in excess, then the rate at which a sin-
coenzyme, it is not surprising that vitamin deficiencies can gle substrate, A, is converted to the product, P, is deter-
have profound pathological effects because of their action mined by the concentration of A:
at the enzyme level.
Other enzymes require monovalent or divalent metal k
A-P
ions as cofactors, generally in a highly selective manner.
The main ions that function as cofactors, along with where k is the rate constant of the reaction. The rate of con-
examples of enzymes that require them, are 1i;ted in version of A to P can be expressed mathematically as
Table 3-9. Especially interesting is Ca2+,whose intracellu-
lar concentration M) is much lower than that of
most other common physiologically important ions. Unlike
Mg2+,Na+, K+, and other cofactor ions, which generally
are present in nonlimiting concentrations, Ca2+is present
in which [A] is the instantaneous concentration of the sub-
strate, k is the rate constant of the reaction, and d [Alldt is
the rate at which A is converted to P with respect to time.
TABLE 3-9
Metal ions that function as cofactors The disappearance of A and the appearance of P are plot-
ted as functions of time in Figure 3-45. Note that the con-
Metal ion Some enzymes requiring metal cofador
centration of A decreases exponentially as the concentra-
Ca2+ Phosphod~esterase tion of P increases exponentially. An exponential time
Prote~nklnase C function is always generated when the rate of change of a
Troponln quantity (d[Alldt in this example) is proportional to the in-
Cu2+(Cu+) Cytochrome oxldase stantaneous value of that quantity ([A] in this example).
Tyroslnase The relationship expressed by Equation 3-10 is more
Fez+or Fe3+ Catalase usefully presented as
Cytochromes
Ferredoxln a - kit
h
log -- -
Perox~dase
a -x 2.303
K+ Pyruvate phosphoklnase(also requlres Mg+2)
Mg2+ Phosphohydrolases
Phosphotransferases c
0
.-
Mn2+ Arglnase s
+
4-4
Phosphotransferases c
(U
0
Na+ Plasma membrane ATPase (also requlres Kt c
0
and Mg2+) 0
Zn2+ Alcohol dehydrcgenase
Carbon~canhydrase T~rne
Carboxypept~dase
B Zero order
f
K,
Substrate
concentration
Figure 3-46 The kinetic order of enzymatic reactions is revealed in
graphical plots. In these plots, the x represents the amount of substrate
A reacting within time t, and a represents the initial amount of A at time
reaction S -
Figure 3-47 At a given enzyme concentration, the initial rate, v,, of the
P rises linearly with increasing substrate concentration.
Eventually all of the enzyme becomes saturated, at which time the en-
zero. The slope is proportional t o the rate constant. (A) Because a first- zyme (E) becomes rate limiting. The Michaelis-Menten constant, KM,is
order reaction has an exponential time course (see Figure 3-45), a plot of equal to the substrate concentration at which the reaction rate is one-half
log (ala - x) versus tgives a straight line. (B) When the enzyme concen- maximum.The black and red curves are for different enzyme concentra-
tration is limiting, the reaction exhibits zero-order kinetics, which gives a tions. Note that KMis independent of the enzyme concentration, [El,
straight line in plots of xversus t. whereas V,, depends directly on [El.
74 PRINCIPLES OF PHYSIOLOGY
......................................
where vo is the initial reaction rate at substrate concentra-
tion [S], Vmaxis the reaction rate with excess substrate, and
K, is the Michaelis-Mentenconstant. Consider the special
case when vo = t Vmx. Substituting for v, ,we get
Dividing by Vmaxgives
Intercept = --
KM I
On rearranging, we obtain
K, + [S] = 2 [S] Figure 3-48 In a Lineweaver-Burk plot, the reciprocal of the reaction
rate, l/vo, is plotted against the reciprocal of the substrate concentration,
l/[S]. For an enzymatic reaction with first-order kinetics, this plot inter-
cepts the horizontal axis at -I/&, and the vertical axis at IIV,,,.
Therefore, K, equals the substrate concentration at which two values of [S]) are needed to construct a Lineweaver-
the initial reaction rate is half what it would be if the sub- Burk plot. V, and K, can be determined from the two
strate were present to saturation. intercepts.
Thus, the Michaelis-Menten constant, K, (in units of Note that the Michaelis-Menten analysis is not limited
moles per liter), depends on the affinity of the enzyme for to enzyme-substrateinteractions, but can be (and often is)
a substrate. For a given enzyme and substrate, K, is equal applied to any system that displays hyperbolic saturation
to the substrate concentration at which the initial reaction kinetics as illustrated in Figure 3-47.
' 1
is r Vmax. By inference, then, K, represents the concentra-
tion of substrate at which half the total enzyme present is Enzyme Inhibition
combined with substrate in the ES; that is, [E,]/[ES] = 2. The activity of most enzymes can be inhibited by certain
The value of K, can be determined from a plot of vo molecules, and as we'll see in the next section, this property
versus [S], as illustrated in Figure 3-47. The greater the of enzymes is used in the living cell as a means of control-
affinity between an enzyme and its substrate, the lower the ling enzymatic reactions. By studying the molecular mech-
K, of the enzyme-substrate interaction. Stated inversely, anisms of inhibition, using both physiological and non-
l/KMis a measure of the affinity of the enzyme for its sub- physiological inhibitors, enzymologists have discovered
strate. As illustrated by the plots for two enzyme con- important features of the active site of enzymes and of the
centrations in Figure 3-47, KM is independent of the en- mechanism of enzyme action. The therapeutic effect of
zyme concentration, whereas Vmaxis dependent on enzyme many drugs depends on their ability to inhibit specific en-
concentration. zymes, thereby blocking metabolic or physiologic processes
The relationship between v, and substrate concen- involved in disease. For example, the drug saralasin blocks
tration described by the Michaelis-Menten equation the action of the enzyme angiotensin I1 and helps lower
(Equation 3-11) is a hyperbolic function. For this reason, blood pressure in humans with severe hypertension.
numerous data points are required to accurately plot vo Enzymes can be poisoned by agents that form highly
against [S] as in Figure 3-47. The Michaelis-Menten equa- stable covalent bonds with groups inside the active site and
tion can be algebraically transformed, however, into a lin- thereby block formation of the enzyme-substrate complex,
ear form called the Lineweaver-Burk equation: ES. Such agents can produce irreversible inhibition, which
cannot be alleviated by removal of the inhibitor; that is, the
1
--- K~ +- 1 (3-16) enzyme is, in effect, rendered permanently inactive. More
vo Vmax ['I Vmax relevant to normal cell function, however, are two types of
reversible inhibition:
It's clear from this equation that a plot of llv, versus 1/[S]
will give a straight line with a slope of K,IVm, and with in- Competitive inhibition is caused by substances that
tercepts of l/Vmaxon the vertical axis and - 1/K, on the appear to react directly with the active site of the en-
horizontal axis (Figure 3-48). Because of the linear nature zyme; it can be reversed by an increase in substrate
of this curve, only two experimental data points (i.e., v, at concentration.
MOLECULES, ENERGY, A N D BIOSYNTHESIS 75
........................................
Noncompetitive inhibition appears to be caused by sub- A Competitive inhibition
stances that bind to a region(~) of the enzyme outside of
the active site; it is not reversed by an increase in sub- Slope of inhibited
strate concentration, but can be reversed by dilution or reaction /!I
removal of the competitor.
Substrate Competitive
inhibitor
SG2 SG3
+ -' "'
/
A Allosteric inhibition
A
A I
Corepressor
C B Allosteric activation
S
w
C
0
0
L
Time
A -----
E,
-
E2 E3 Ed E5
End product
As mentioned earlier, carbohydrates, lipids, and pro-
teins ingested in foodstuffs are the primary fuels for ani-
mals. After digestion, these molecules generally enter the
circulatory system as five- or six-carbon sugars, fatty acids,
Inhibition
and amino acids, respectively (Figure 3-56). These small
molecules then enter the tissues and cells of the animal,
Figure 3-54 Products of a reaction sequence can allostericallyinhibit a where they may (1)be immediately broken down into
rate-limiting enzyme in the pathway. The metabolic end product in this Smallermolecules for the extraction of chemical energy or
case directly affects the activity of the initial enzyme, as illustrated in Fig-
ure 3-53A. In contrast, in the mechanism depicted in Figure 3-52 the end
for rearrangement and recombination into other types of
product affects the amount of enzyme by controlling transcription of its molecules, or (2)be up larger such as
gene. polysaccharides (e.g., glycogen),fats, or proteins. With few
MOLECULES, ENERGY, A N D BIOSYNTHESIS 79
......................................................................
0, Fuel Figure 3-55 The hydrolysis of ATP
powers numerous energy-requiring
processes in biological systems. The
ADP produced by. hydrolysis
. . is recy-
cled to ATP by rephosphorylation en-
ergized by the oxidation of foodstuff
+ P, I)
€
ADP molecules to CO, and H20.
3-carbon
sugars
Citric
acid cycle
Electron
transport
80 PRINCIPLES OF PHYSIOLOGY
.............................................................
exceptions, these too will eventually be broken down and Outer membrane
eliminated as C 0 2 , H20, and urea. Nearly all molecular
constituents of a cell are in dynamic equilibrium, constantly
I
being replaced by components newly synthesized from sim-
pler organic molecules.
Some simple organisms, including certain bacteria and
yeasts, as well as a few invertebrate species, can live indef-
initely under totally anaerobic (i.e., essentially oxygen-free)
conditions. The anaerobes fall into two groups: obligatory
anaerobes, which cannot grow in the presence of oxygen
(e.g., botulism bacterium, Clostridium botulinum), and fac-
ultative anaerobes, which survive and reproduce well either
in the absence or in the presence of oxygen (e.g., yeasts).All
vertebrates and most invertebrates, however, require mole-
cular oxygen for cellular respiration and are therefore
termed aerobic. Even these aerobic animals generally pos-
sess tissues that can metabolize anaerobically for periods of
time, building up an oxygen debt that is repaid when suf-
ficient oxygen becomes available.
As these observations suggest, there are two kinds
of energy-yielding metabolic pathways in animal tissues
(Figure 3-57):
-
and water by combustion in the overall reaction an oxidant. The free-energy change in each reaction is pro-
portional to the difference between the electron pressures
C,H,,O, + 6 0, 6 CO, + 6 H20 of the two molecules of the redox pair.
In aerobic cell metabolism, electrons move sequentially
AGO = -686 kcal.mo1-'
from compounds of higher electron pressure to com-
pounds of lower electron pressure. The final electron ac-
The 686 kcal liberated by the oxidation of 1 mol of glu-
ceptor in aerobic metabolism is molecular oxygen. Since
cose is the difference between the free energy incorporated
oxygen acts merely as an electron acceptor, it is possible in
into the structure of the glucose molecule during photo-
theory to support aerobic metabolism without oxygen,
synthesis and the total free energy contained in the carbon
provided that a suitable electron acceptor is supplied in
dioxide and water produced. If 1 mol of glucose is oxi-
place of oxygen.
dized to carbon dioxide and water in a one-step combus-
In being transferred from glucose to oxygen, electrons
tion (i.e., if it is burned), the free-energy change will appear
undergo an enormous drop in both reduction potential and
simply as 686 kcal of heat. During cellular respiration,
free energy. One of the functions of cell metabolism is to
however, a portion of this energy, instead of appearing as
transport electrons from glucose to oxygen in a series of
heat, is conserved as useful chemical energy and is chan-
small steps instead of one large drop. This transport is im-
neled into ATP through the phosphorylation of ADP. The
plemented by two mechanisms found in all cells. First, as
overall reaction for the metabolic oxidation of glucose by
we noted earlier, the chemical conversion of foodstuff mol-
the cell, including the coupled conversion of ADP to ATP,
ecules such as glucose to the fully oxidized end products
can be written as
C6Hl,0, + 38 P, + 38 ADP + 6 0,
6 CO,
-
+ 6 H 2 0 + 38 ATP
(e.g., CO, and H,O) occurs in steps and involves many in-
termediate compounds. Second, electrons removed from
substrate molecules are passed to oxygen via a series of
electron acceptors and donors of progressively lower elec-
AGO = -420 kcal (as heat) tron pressure. As we will see shortly, these mechanisms al-
low energy to be channeled into the synthesis of ATP in
Thus, 266 kcal(686 - 420) is incorporated into 38 mol of "packets" of appropriate size.
ATP (7kcal .molkl ATP).
How is the free energy of the glucose molecule trans-
ferred to ATP? To understand this, we must first recall that
What reasons can you thlnk of t o explain why
oxidation of a molecule is most broadly defined as the
anlrnal life evolved with 0, as the final electron
transfer of electrons from that molecule to another mole-
acceptor?
cule; conversely, reduction of a molecule is the acceptance
" . A -.. . m lWY r A..%.*
r>ri-,w &.A- r- ..Li
of electrons from another molecule. In an oxidation-
reduction reaction the reductant (electron donor) is oxi-
dized by the oxidant (electron acceptor). Together they Electron-transferring coenzymes
form a redox pair: During certain biochemical reactions, electrons, together
with protons (i.e., hydrogens), are removed from substrate
electron donor e- + electron acceptor molecules by enzymes collectively called dehydrogenases.
These enzymes all function in conjunction with pyridine or
flavin coenzymes. The most common are nicotinamide ade-
nine dinucleotide (NAD+),noted earlier, and flavin adenine
reductant n e- + oxidant dinucleotide (FAD).Their structural formulas are shown in
Figure 3-59. These coenzymes act as electron acceptors in
'
where n is the number of electrons transferred. Whenever their oxidized form and as electron donors in their reduced
electrons are accepted from a reductant by an oxidant, en- form:
82 PRINCIPLES OF PHYSIOLOGY
..............................................................................
A FAD Figure 3-59 The two most common elec-
tron-carrying coenzymes-flavin adenine
Adenosine dinucleotide (FAD) and nicotinamide ade-
nine dinucleotide (NAD+)-conta~n an
adenosine group and a vitamin-derived
group (color shading). Riboflavin, one of the
Adenosine
B vitamins, is part of FAD, and nicotinamide,
one form of the vitamin niacin, is part of
NAD+. In the oxidized coenzyme molecules,
shown here, the atoms in red type can ac-
cept protons and electrons.
Riboflavin
reduced substrate + NAD+ L- 2 mol of reduced FAD are produced. Thus, of the 686 kcal
NADH + H + + oxidized substrate of free energy available from the oxidation of 1rnol of glu-
cose, about 624 kcal (12 x 52), or 91%, is transferred
reduced substrate + FAD ==T to the electron-transferring coenzymes to be released in
FADH2 + oxidized substrate subsequent stages of electron transfer. As noted earlier,
266 kcal of this free energy is eventually retained by the
A very convenient property of these coenzymes for experi- synthesis of ATP.
mental purposes is that their reduced and oxidized forms
have different ultraviolet absorption spectra (Figure 3-60). Electron-transport chain
They also undergo a change in ultraviolet-excited fluores- In spite of the large difference in electron pressure between
cence upon oxidation and reduction. These two properties NADH or FADH, and oxygen, there is no enzymatic mech-
have permitted physiologists and biochemists to use pho- anism by which these reduced coenzymes can be directly
tometric methods to monitor changes in the amount of re- oxidized by oxygen. Instead, an elaborate electron-trans-
duced coenzyme under experimental conditions in living port chain, or respiratory chain, has evolved in which the
cells. electrons move through about seven discrete steps from the
The free energy of the reduced forms of both coen- high reducing potential of NADH and FADH, to the final
zymes, NADH and FADH,, is very high relative to oxygen. electron acceptor, molecular oxygen. This sequence of elec-
As a result, the transfer of electrons from the reduced tron transfers is the final common pathway for all electrons
coenzymes to oxygen is accompanied by a large decrease during aerobic metabolism. Its function, as we will see, is
NADH + 4 0, -
in free energy. For example, the A G O for the reaction
NAD+ + H,O, in which two elec-
trons are transferred from NADH to O,, is about
to utilize the energy of electron transfer efficiently for the
phosphorylation of ADP to ATP.
The electron-transport chain consists of a series of pro-
-52 kcal .molk'. The AGO for transfer of electrons from teins and coenzymes that can exist in oxidized and reduced
FADH, to oxygen is about the same. During the oxidation forms. Several of the electron carriers in the chain are iron-
of 1mol of glucose in cells, 10 mol of reduced NAD and containing proteins called cytochromes, each of which con-
I. Figure 3-60 The oxidized and reduced forms of
I ;Oxidized NAD+ have different absorption spectra. Since the
CONH, difference in absorption is greatest at 340 nm, that
wavelength is used to monitor the reduction of
NAD+ t o NADH, and vice versa. [Adapted from
I Lehninger, 1975.1
R
NADH. reduced form
tains a substituted heme group. The deeply colored heme trons are transferred from NADH down the electron-trans-
group consists, essentially, of a porphyrin ring with an iron port chain in a series of coupled reactions, ending with the
atom at its center; the heme group is also present in the he- reduction of molecular oxygen. Only the last enzyme in the
moglobin molecule of vertebrate red blood cells (see chain, cytochrome aa3, is able to transfer its electrons di-
Figure 13-2B).The substituted heme groups in the various rectly to oxygen. The order in which the various electron
cytochromes differ in the side chains attached to the por- carriers function was worked out in studies with several poi-
phyrin ring (Figure 3-61).The cytochromes show charac- sons that block the flow of electrons at specific points in the
teristic absorption spectra in their oxidized and reduced chain. In these studies, the presence of the oxidized or re-
forms, absorbing more strongly in the red when reduced. duced forms of the electron carriers was monitored by spec-
This behavior led to the first discovery of their function by trophotometric methods. For example, when the final
David Keilin in 1925. Using a spectroscope, he discovered step-electron transfer by cytochrome oxidase (composed
that the flight muscles of insects contain compounds that of subunits of a and a3)to 0,-is blocked by cyanide, the
are oxidized and reduced during respiration. He named effect on electron transport is identical to the removal of
these compounds cytochromes and hypothesized that they molecular oxygen (see Figure 3-62). Electrons pile up, so to
transfer electrons from energy-rich substrates to oxygen. speak, because transport is interrupted along the chain, re-
The functional order of the components of the electron- ducing all the cytochromes and other electron carriers above
transport chain is diagrammed in Figure 3-62. From left to the point of the block. Another poison, antimycin, blocks
right in this figure, each successive electron carrier has a the flow of electrons from cytochrome b to c, causing the
lower electron pressure than its predecessor. As a result, elec- electron carriers above the block to become fully reduced
and those below the block to become completely oxidized.
The cascading of electrons through a series of small dis-
crete steps, contrasted with the direct reduction of oxygen
by NADH or FADH,, confers a great energetic advantage
on the cell. The "logic" of the electron-transport system be-
comes apparent when it is recalled that the energy required
to convert ADP to ATP, the standard currency in biological
energy exchange, is small compared with the total change
in free energy produced by the transfer of electrons from
NADH to oxygen. As we saw earlier, it requires a mini-
mum of 7.3 kcal to synthesize ATP from ADP and inor-
ganic phosphate, whereas 52 kcal are released in the trans-
fer of two electrons from NADH to oxygen. Oxidation
of NADH in a one-step reaction could be coupled to the
formation of only one molecule of ATP. Such a coupled
reaction would be quite inefficient, conserving only 14%
(7.3152)of the available free energy in NADH in the form
of ATP, the rest being lost as heat. In contrast, the electron-
Figure 3-61 Heme Aacts as the electron acceptor-donator group of cy-
tochrome aa,. At the center of the porphyrin ring is the iron atom that is
transport system is a much more efficient mechanism that
oxidized or reduced during transport. The side groups highlighted in releases the total energy available in NADH (and FADH, )
color differ in the other cytochromes. in small doses just large enough for synthesis of ATP.
PRINCIPLES OF PHYSIOLOGY
--- - - - - - - - - Flavo-
protein
52
kcal
Figure 3-62 In the electron-transport cham, electrons cascade from quence, were useful In determining the order In which the electron car-
one carrier to the next In order of their decreasing electron pressures rlers funaion
Resp~ratorypolsons (red shad~ng),which blockx specific steps In th~sse-
The actual synthesis of ATP from ADP and inorganic oxygen atoms (40, ) consumed in the formation of H,O.
phosphate (PI)during electron transport is called oxidative Thus, PI0 = 3. Reduced FAD, however, transfers electrons
phosphorylation, or respiratory-chain phosphorylation. directly to coenzyme Q, bypassing the first phosphorylation
The phosphorylation of ADP to ATP occurs as a conse- reaction; thus, for each pair of electrons transferred from
quence of the transfer of electrons between three pairs of FADH, to atomic oxygen, only two ATP molecules are
electron carriers: formed, for a PI0 = 2.
Of the several theories proposed to explain how the
Flavoprotein to coenzyme Q synthesis of ATP is coupled at the molecular level to the free
energy liberated during electron transfer, the chemiosmotic
Cytochrome b to cytochromes c and c, theory of energy transduction is the most widely accepted.
Cytochromes aa, (cytochrome oxidase) to molecular This mechanism is discussed in Chapter 4. It is interesting
ovgen to note here that oxidative phosphorylation becomes un-
coupled from electron transport whenever anything hap-
At each of these three steps in the electron-transport chain, pens that makes the inner mitochondria1 membrane
the drop in free energy is adequate to drive the phospho- "leaky," that is, more permeable than normal to H+ or
rylation of ADP (Figure 3-63). Thus, for each pair of elec- other cations. In this case, the production of ATP drops or
trons that passes along the entire chain, three molecules of ceases while both electron transport and the reduction of
ATP are generated from three molecules of ADP and three 0, to H,O continue. All the released energy is given off as
molecules of PI. Each pair of electrons finally reduces one- heat. Oxidative phosphorylation is also uncoupled from
half molecule of 0, to form one molecule of water: electron transport by certain drugs, such as dinitrophenol
(DNP).Because this drug reduces the efficiency of energy
metabolism, it was once prescribed by physicians to help
patients lose weight. Its use as a weight-reducing drug was
discontinued when it was found to produce pathological
By comparing the amount of 0, consumed (i.e., con- side effects.
verted to water) and the amount of Pi consumed (i.e., in-
corporated into ATP), we can establish the P I 0 ratio (ratio
of Pi to atomic oxygen).For example, if one oxidative phos- GIycolysis
phorylation occurs at each of the three steps noted above, The term glycolysis, which means "breakdown of sugar,"
3 mol of Pi will be incorporated into ATP for each mol of refers to the sequence of reactions by which the six-carbon
NAD 2e- Q 2e- CC, 2e- H2O
+
+02 2H+
Figure 3-63 As two electronsfrom a molecule of NADH flow through the protein; Q, coenzyme Q. The letters b, c, c, , a, and a, refer to the re-
entire electron-transport chain, three molecules of ADP are phosphory- spective cytochromes, shown working in pairs transporting pairs of elec-
lated t o ATP. Formation of ATP occurs at the indicated steps. FP, flavo- trons.
sugar glucose is converted to pyruvic acid, a three-carbon acetone phosphate. The latter molecule is enzymatically re-
molecule (see Figure 3-56). This sequence of reactions, the arranged into the former, so that each mole of glucose
most fundamental in the energy metabolism of animal cells, yields 2 rnol of glyceraldehyde 3-phosphate. This completes
is required for both anaerobic and aerobic release of energy the first stage of glycolysis, in which 1 rnol of the six-carbon
from foodstuffs. The glycolytic pathway is also called the glucose is converted into 2 rnol of the three-carbon glycer-
Embden-Meyerhof pathway after the two German bio- aldehyde 3-phosphate (steps 1-5 in Figure 3-64).
chemists who worked out the details of glycolysis in the The second stage of glycolysis begins with the oxida-
1930s. tion of glyceraldehyde 3-phosphate and addition of a phos-
The first step in the glycolytic pathway is the phospho- phate group to yield 1,3-diphosphoglycerate (step 6). In
rylation of glucose by ATP to give glucose 6-phosphate this crucial reaction the energy that would otherwise be re-
(Figure 3-64). The breakdown of glycogen also yields phos- leased by oxidation of the aldehyde group is captured in a
phorylated glucose, which then can enter the glycolytic high-energy acyl-phosphate bond linking the second phos-
pathway (see Figure 9-13). Glucose 6-phosphate then is phate group to the carbonyl carbon atom (see Figure 3-39).
converted to fructose 6-phosphate, which is phosphoryl- The elucidation of the mechanism of this reaction and of
ated to fructose l,6-diphosphate at the expense of a second the following one (step 7), in which ADP is directly phos-
ATP (steps 2 and 3). At first glance, it would seem uneco- phorylated to ATP by the substrate, is considered among
nomical for the cell to expend 2 mol of ATP to phospho- the most important contributions to modern biology.
rylate 1rnol of hexose, since the object of glycolysis is to Through these discoveries, Otto Warburg and his col-
produce ATP. On closer examination, however, the phos- leagues provided, in the late 1930s, the first insight into a
phorylation of glucose does make sense. At physiological mechanism by which chemical energy of oxidation is con-
pH, phosphorylated hexose and triose (three-carbon sugar) served in the form of ATP. This process is called substrate-
molecules become ionized and thus have very low mem- level phosphorylation to distinguish it from the process of
brane permeabilities. Although unphosphorylated glucose oxidative phosphorylation coupled to electron flow in the
is free to enter (or leave) the cell by diffusion through respiratory chain.
the surface membrane, the phosphorylated form is con- In steps 8-10 of glycolysis, 3-phosphoglyceric acid is
veniently trapped along with its phosphorylated deriva- converted to 2-phosphoglyceric acid, water is removed to
tives within the cell. The 2 mol of ATP expended in these form phosphoenolpyruvate, and finally the latter yields its
so-called priming phosphorylations is, in fact, not really phosphate group to ADP, forming ATP and pyruvic acid,
lost, for later in the glycolytic pathway these phos- another substrate-level phosphorylation. Thus, the gly-
phate groups-and their intramolecular free energies-are colytic pathway ends with 2 rnol of pyruvic acid produced
used to regenerate ATP, thereby conserving the energy of
ATP ATP from each mole of glucose. The phosphorylation of each
ATP
the phosphate groups utilized in the priming phosphoryla- mole of hexose consumes 2 rnol of ATP, and each mole of
tions. triose generates 2 rnol of ATP (steps 7 and 10).Since each
Fructose 1,6-diphosphate is cleaved in step 4 into two mole of glucose yields 2 rnol of triose, the net gain per mole
triose sugars, glyceraldehyde 3-phosphate and dihydroxy- of glucose in glycolysis is 2 rnol of ATP.
86 PRINCIPLES O F PHYSIOLOGY
..............................................................................
Fructose 1,6-diphosphate
I,
CH2-0-P-0- CH,-OH
I
I
CH-OH
0-
-C=O
@
H-C=O
I
CH,-0-P-0-
ii
I
Dlhydroxyacetone phosphate
I L I
ADP COOH
3-Phosphoglyceric acid
2-Phosphoglyceric acid
F"' ii
C-O-P-O-
ADP
iL,
@
P"
c=o
I
COOH
I
0-
COOH
Phosphoenolpyruvic acid Pyruvic acld
NADH 7j
Figure 3-64 In the Embden-Meyerhof glycolytlc pathway, glucose IS de-
graded to pyruvlc acld Under aerobic conditions, pyruvlc acld IS further CH,-OH
oxldlzed, In the absence of oxygen, however, ~tIS reduced t o lactate
Note that at step 4 each hexose molecule IS split into two trlose mole- HO-C-H
cules Thls doubles the molarltles of reactants In the remainder of the I
pathway The common energy intermed~ates,ATP and NADH, are HOOC
shaded. Lactic acid
As indicated in Figure 3-65, glycolysis of 1mol of glu-
cose also yields 2 mol of NADH. In the presence of oxy- 2ATP
gen-that is, during aerobic metabolism-each mole of
NADH eventually is oxidized by molecular oxygen, via 2ATP
the electron-transport chain, with concomitant produc-
tion of 3 mol of ATP (see Figure 3-63). In this way, NAD+
is regenerated for use in the glycolytic pathway. In the ab- 0
sence of oxygen-that is, during anaerobic metabolism-
pyruvic acid resulting from glycolysis is reduced to lactate Pyruvic acid
(Figure 3-64, step 11)or, in certain microorganisms such
as yeast, to ethanol. This substrate reduction is coupled to
0
oxidation of NADH, thereby replenishing the NAD+ Figure 3-66 Under anaerobic conditions, the NADi consumed in the
reduced to NADH in step 6 of glycolysis. In this case the
electrons of NADH are accepted by pyruvate instead of
oxygen. Without this anaerobic oxidation of the reduced
-
breakdown of glucose to pyruvic acid is replenishedby reduct~onof pyru-
vic acid to lactate. This NAD+ NADH cycle permits glycolysis to
2ATP
proceed in the absence of oxygen. Circled numbers refer to steps in gly-
colytic pathway shown in Figure 3-64.
coenzyme, there would be a depletion of the oxidized
form of the coenzyme (NAD+),and glycolysis would be
blocked for lack of an electron acceptor at step 6 (the All the reactions of the glycolytic pathway leading to
oxidation of 3-phosphoglyceraldehyde to 1,3-diphospho- formation of pyruvic acid occur in free solution in the cy-
glycerate) in the absence of molecular oxygen. The anaer- tosol. The formation of acetyl CoA and CO, from pyruvic
obic NAD+ NADH cycle that operates between acid and the eight major reactions composing the citric acid
steps 6 and 11 of the glycolytic pathway is shown in cycle are all catalyzed by enzymes confined to the matrix
Figure 3-66. compartment or inner membrane of mitochondria. For
each acetate residue that enters the cycle, two additional
Citric Acid Cycle molecules of CO, and two molecules of H,O are produced.
Under aerobic conditions, pyruvic acid is decarboxylated The overall reaction for the complete oxidation of pyruvate
(i.e., 1 mol of CO, is removed), leaving a two-carbon via the citric acid cycle and electron-transport chain is writ-
acetate residue, which then reacts with coenzyme A (CoA)
-
ten as follows:
to yield acetyl coenzyme A (acetyl CoA). In this coupled re-
action, NAD+ accepts one hydrogen atom from pyruvic 2 CH,COCOOH + 5 0, 6 CO, + 4 H,O
acid and one from coenzyme A (Figure 3-67). Coenzyme A
acts as a carrier for the acetate residue, transferring it to The citric acid cycle is also known as the Krebs cycle af-
oxaloacetic acid to form citric acid in the first reaction of ter Hans Krebs, who in the early 1940s elucidated the ma-
the cimc acid cycle (Figure3-68). This reaction releases free jor features of the reaction sequence and its cyclic nature.
CoA, which is recycled to repeatedly transfer acetate (It's also known as the tricarboxylic acid cycle because sev-
residues from pyruvic acid to oxaloacetate. eral intermediates have three carboxyl groups.) The two-
carbon acetate residue of acetyl CoA first condenses with
the four-carbon oxaloacetic acid to form the six-carbon cit-
Glucose ric acid (see Figure 3-68, step 1).In steps 4 and 5, two car-
boxyl groups of isocitric acid are removed to form two
--l-2ADp
molecules of CO,. Moreover, four hydrogen atoms are
transferred to NAD+ to form two molecules of NADH.
k - 7-
Fructose 1,6-d~phosphate
Step 6 is catalyzed by succinic dehydrogenase, which is
b bound to the inner mitochondria1 membrane. In this reac-
tion, two hydrogen atoms are transferred from succinic
2,3-Phosphoglyceraldehyde Dihydroxyacetonephosphate
--
2 P,-
2 NAD+ P NARH
+
NADH
ZADP- -= COA - SH
-rn
COOH S- CoA
2ADP- Pyruvic acid Acetyl coenzyme A
Y
2 Pyruvlc ac~d Figure 3-67 Pyruvic acid is decarboxylated and its two-carbon acetyl
group (shaded) is transferredto coenzyme A (CoA) in a coupled reaction
Figure 3-65 ATP IS both consumed and produced durlng glycolys~sOx- that yields acetyl CoA. This reaction occurs in the matrix compartment of
ylelds a net of 2 rnol of ATP as
ldatlon of 1 rnol of glucose to pyruvlc ac~d mitochondria; it links glycolysis, which occurs in the cytosol, with cellular
well as 2 mol of NADH. respiration within mitochondria.
88 PRINCIPLES O F PHYSIOLOGY
..............................................................................
COOH
I 5OOH
I
C Hz C-COOH
f CoA-SH \ I
7HO--F-COOH I/
I
0 COOH
I1 COOH cis-Asconitic acid I
COOH k OOH
CH,-C-COOH Citric acid
Pyruvic acid Acetyl CoA +
H,O
Oxaloacetic acid CI OOH
lsocitric acid
7I H2
I
C=O GDP + P, I
CHZ
a-Keto-
I
glutaric
COOH
7
\C OOH
I
Succinic acid
COOH I I
CH
Malic acid 11
I
COOH
Fumaric acid
Figure 3-68 In the course of each circuit of the citric acid (Krebs)cycle, four pairs of protons (shaded)to electron-carrier coenzymes. The carbons
an acetate group transferred from acetyl CoA moves through several in- of each entering acetyl group (red type) remain intact during its initial cir-
terrnediates with the production of two CO, molecules and transfer of cuit around the cycle. Note that one molecule of GTP is produced.
acid to FAD, forming fumaric acid and FADH, . Another Steam engines have attained efficiencies of approximately
oxidation occurs when malic acid is converted to ox- 30%. Now let us see how efficiently the living cell transfers
aloacetic acid by the transfer of two hydrogen atoms to chemical energy from glucose to ATP.
NAD+ (step 8). A new acetate residue then condenses with Under standard conditions, it takes about 7 kcal to
the oxaloacetate to reconstitute the citric acid molecule, phosphorylate 1mol of ADP to form ATP. If the free energy
thus beginning another repetition of the cycle. of glucose were conserved with an efficiency of loo%, each
Each time one circuit of the citric acid cycle is com- mole of glucose could energize the synthesis of 98 mol of
pleted, two carbon atoms and four oxygen atoms are re- ATP (68617 = 98) from ADP and inorganic phosphate. In
moved as two molecules of CO, and eight hydrogen atoms fact, during the metabolic oxidation of 1 rnol of glucose
are removed, two at a time (Figure 3-69). These hydro-
GTP only 38 mol of ATP are synthesized, giving an overall effi-
gens (as electrons accompanied by protons), carried by ciency of about 42% or more.'' The remaining free energy
NADH and FADH,, are fed into the respiratory chain and is liberated as metabolic heat, which accounts for a part of
eventually oxidized to H,O by molecular oxygen (see the heat that warms and thereby increases the metabolic
Figure 3-62). The CO, leaves the mitochondrion and rate of the tissue. Essentially all the energy incorporated
then the cell by simple diffusion; it is finally eliminated as into ATP and transferred to other molecules is eventually
a gas via the circulatory and respiratory systems (see degraded to heat. The oxidation of fossil fuels represents
Chapter 13). a long-delayed return of stored energy to the original low-
energy, high-entropy state of CO, and water.
Efficiency of Energy Metabolism
The direct oxidation (burning)of glucose and its metabolic
oxidation both liberate the same of free energy- * The 42% calculated here is for standard conditions. The efficiency of en-
namely, 686 kcal .mol-'. If water is boiled by the heat of ergy conservation by the cell may in fact be as high as 60%, because the
free energy of hydrolysis of ATP under intracellular conditions has been
burning glucose produce pressure for a en- estimated to be greater than that under standard conditions. The ener-
gine, the mechanical output of the engine divided by the getic efficiency of ATP production is therefore substantially better than
that of a steam engine, in fact better than that of any other method
free-energy drop of 686 kcal. m o l - ~represents the effi- yet devised by humans for converting chemical energy to mechanical
ciency of the conversion of chemical to mechanical energy. energy.
Figure 3-68). This adds up to 38 rnol of nucleotide
I /-"iA
Pyruvic acid
ICO,I* triphosphate per mole of glucose during aerobic respira-
tion. As noted earlier, only 2 rnol is produced during
anaerobic respiration. Thus, although aerobic respiration
conserves a minimum of about 42% of the free energy of
the glucose molecule, anaerobic respiration conserves
only about 2%. Stated differently, the energy conserva-
tion of glucose metabolism via aerobic glycolysis and the
citric acid cycle is about 20 times as efficient as that via
anaerobic glycolysis. It is not surprising, then, that most
animals carry on aerobic metabolism and require molec-
ular oxygen for survival.
Oxygen Debt
When animal tissue, such as active muscle, receives less
oxygen than is required to produce adequate amounts of
ATP by respiratory-chain phosphorylation, some of the
pyruvic acid, instead of going on to fuel the citric acid cy-
cle, is reduced to lactic acid. For every 2 rnol of pyruvic acid
reduced, 2 rnol of NADH is oxidized, costing 6 rnol of ATP
that might have been synthesized by respiratory-chain
phosphorylation. If the oxygen deficiency is maintained,
Figure 3-69 The oxidation of pyruvic acid produces CO,, NADH, lactic acid concentrations rise, and some may enter the ex-
FADH,, and GTP. The CO,, a waste product, is eliminated. Both NADH
tracellular space and circulatory system. When the muscle
and FADH, enter the electron-transport chain where the~relectron pres-
sure is converted into ATP during oxidative phosphorylation (see Figure
stops its strenuous activity and oxygen is again available,
3-63). GTP is formed in a substrate-level phosphorylationand like ATP can the enzyme lactate dehydrogenase oxidizes the accumu-
drive energy-requiring reactions. The circled numbers refer to the corre- lated lactic acid back to pyruvic acid coupled to the reduc-
sponding steps in Figure 3-68. [Adapted from Vander et al., 1975.1 tion of NAD+ (Figure 3-70).The NADH produced in this
reaction then is oxidized in the respiratory chain, recoup-
ing some of the ATP forfeited by the anaerobic formation
of lactic acid. Moreover, some of the pyruvic acid regener-
It is interesting to compare the efficiencies of anaerobic ated from lactic acid goes on to fuel the citric acid cycle,
and aerobic glucose metabolism, keeping in mind that eventually recouping more of the forfeited ATP. (Some of
since each mole of glucose yields 2 rnol of the three-carbon the recovered pyruvic acid may be used for the synthesis of
derivatives, it is necessary to double all the molarities be- alanine and glucose.)
yond step 5 of glycolysis. In anaerobic glycolysis there is a In other words, when muscle is starved for oxygen,
GTP
net production of 2 rnol of ATP per mole of glucose (see it switches to anaerobic metabolism in which ATP is
Figure 3-65), because 2 of the 4 rnol of ATP produced by formed with low efficiency. But the unused chemical en-
substrate-level phosphorylation of ADP are consumed in ergy is stored in the tissue as lactic acid, and later becomes
the priming phosphorylations. As noted previously, the available for aerobic metabolism when sufficient oxygen
2 rnol of NADH produced by the oxidation of 3-phospho- is available. With cessation of heavy exercise, the respira-
glyceraldehydeis reoxidized to NAD+ when the two pairs tory and circulatory systems continue for some time to
of hydrogen atoms are transferred to 2 rnol of pyruvic acid supply large amounts of oxygen in order to "repay" the
to form 2 rnol of lactic acid under anaerobic conditions (see oxygen debt that was built up as an accumulation of
Figure 3-66). lactic acid.
Under aerobic conditions, each of the 2 rnol of
NADH produced in glycolysis by the oxidation of 3-
phosphoglyceraldehyde yields 3 rnol of ATP during ox-
idative phosphorylation (Figure 3-63). Pyruvic acid goes OH 0
on to fuel the citric acid cycle, yielding a total of 10 pairs Lactate dehydrogenase I1
CH3-CH-COOH / \ > CH3-C-COOH
of hydrogen atoms for every 2 rnol of pyruvic acid Lactic acid NAD+ NADH Pyruvic acid
(Figure 3-69). Eight pairs are carried by NAD+, yielding
24 rnol of ATP, while two pairs are carried by FAD, yield- Figure 3-70 The lacticacid that accumulates in muscle during strenuous
activity is oxidized to pyruvate following cessation of activity when the
ing 4 rnol of ATP. Finally, 2 rnol of GTP is produced by
oxygen supply increases. Oxidation of the NADH, as well as some of the
substrate-level phosphorylation of guanosine diphos- pyruvate produced in this reaction, eventually fuels respiratory-chain
phate (GDP) during the oxidation of a-ketoglutaric acid phosphorylation, thereby recovering some of the ATP forfeited during
to succinic acid in step 5 of the citric acid cycle (see anaerobic production of lactic acid.
90 PRINCIPLES O F P H Y S I O L O G Y
.........................................
SUMMARY free energy and increasing entropy. Living systems appear
to defy entropy, but they do not; they merely exist at the ex-
Biologists generally accept the hypothesis that life on Earth pense of chemical energy obtained from their environment.
arose spontaneously in shallow seas under special condi- Energy-requiring biological reactions utilize ATP, a
tions that no longer exist. It is believed that organic mole- triply phosphorylated nucleotide that serves as a common
cules, synthesized in the primitive atmosphere by reactions intermediate capable of contributing chemical energy
energized by lightning discharges or radiation, accumulated stored in the form of its terminal phosphate bond. This en-
in the water over long periods, providing the raw material ergy transfer is accomplished by means of coupled reac-
for primordial living cells. tions in which an endergonic (energy-requiring)reaction is
Living matter is composed primarily of carbon, nitro- driven by an exergonic (energy-releasing)reaction. ATP is
gen, oxygen, and hydrogen in stable (covalently bonded) reconstituted from ADP by the oxidation of foodstuff mol-
associations. Carbon, nitrogen, and oxygen are capable of ecules, which largely have their origin in the radiant solar
forming double and triple bonds, which greatly increase the energy trapped during the process of photosynthesis in
structural variety of biological molecules. green plants. Thus, animals depend on energy ultimately
The polarity of the water molecule is responsible for hy- derived from the sun.
drogen bonding. Besides linking hydrogen and oxygen Enzymes act as biological catalysts; that is, they lower
atoms of adjacent water molecules, hydrogen bonding con- the energy required to activate reactants sufficiently for re-
fers on water many special properties that have profoundly action and thereby increase the rate of reaction at a given
shaped the evolution and survival of animal organisms. temperature. With the aid of enzymes, cell chemistry can
Water dissociates spontaneously into H+ and OH-; in proceed at reasonable body temperatures. The catalytic ac-
1 liter of pure water there is lo-' mol of each ion. Many tion of an enzyme arises from its ability to bind specific re-
substances in solution contribute to an imbalance in the actant molecules at the active site; the close steric fit re-
concentrations of H+and OH-, giving- rise to acid-base be-
-
quired for this interaction is largely responsible for enzyme
havior (i.e., donation and acceptance of protons). Their specificity. This binding produces favorable spatial relations
concentrations are measured by the pH system. The pH of between the reacting molecules. Regulation of enzyme con-
biological fluids influences the charges carried by amino centrations appropriate to the needs, function, and envi-
acid side groups and hence the conformation and activity ronment of the cell is performed at the level of DNA
of proteins. Physiological buffering systems maintain the through enzyme induction and repression. The activity of
intra- and extracellular pH within a narrow range. some enzymes can also be controlled by the binding of reg-
The electrostatic force attracting an ion to a site of op- ulatory molecules or ions to the enzyme molecule at an al-
posite charge is determined by the distance of closest ap- losteric site that is distinct from the active site of the en-
proach of the ion to the site. The ion selectivity of a site de- zyme. This binding results in a conformational change that
pends on the relative ability of the site to compete with the affecrs the properties of the active site.
dipolar water molecules in binding different ion species. Liberation of the free energy stored in foodstuffs dur-
Four major groups of organic molecules compose ani- ing metabolism occurs by transfer of electrons from an elec-
mal cells. The lipids, which include triglycerides (fats),fatty tron donor (reducer)to an oxidizer. The release of free en-
acids, waxes, sterols, and phospholipids, are important as ergy in the cell is budgeted in small steps compatible with
energy stores and as constituents of biological membranes. the amounts of free energy required to phosphorylate ADP
The carbohydrates include sugars, storage carbohydrates to ATP. For example, electrons from the reduced coen-
(glycogen and starch), and structural polymers such as zymes NADH and FADH2 are transported in increments
chitin and cellulose. The sugars, glycogen, and starch are down a chain of electron acceptors and electron donors,
major sources of substrate for energy metabolism by cells. yielding enough energy for synthesis of ATP at three points
Proteins, made up of linearly arranged amino acid residues, in the chain. An electron-pressure gradient exists along the
form many structural materials such as collagen, keratin, electron-transport chain of cytochromes, so that electrons
and subcellular fibrils and tubules. Enzymes are specialized flow to the ultimate electron acceptor, molecular oxygen. It
proteins bearing catalytically active sites and are important is the electron-hungry nature of the oxygen atom, and its
in nearly all biological reactions. The nucleic acids DNA abundance on the surface of the Earth, that makes it the
and RNA encode the genetic information necessary for the ideal terminal electron acceptor in living systems.
orderly synthesis of all the protein molecules in the cell. During glycolysis, each glucose molecule is broken
A major characteristic of biological systems is that they down into two molecules of the three-carbon pyruvic acid
maintain a low state of entropy -that is, they are highly with net formation of 2 ATP molecules and 2 NADH mol-
and improbably organized. A steady expenditure of energy ecules. During anaerobic metabolism, the pyruvic acid is re-
must therefore be derived from foodstuff molecules by the duced to lactate, regenerating the NAD+ consumed during
processes of energy metabolism. In the living cell, metabo- glycolysis. During aerobic metabolism, pyruvic acid pro-
lism occurs as orderly, regulated sequences of chemical re- duced by glycolysis is oxidized completely to CO, and
actions catalyzed by enzymes. Chemical reactions sponta- H,O, via the citric acid cycle and respiratory chain. Oxi-
neously tend to go down an energy gradient, decreasing dation of 2 molecules of pyruvic acid is accompanied by the
3LECULES, ENERGY, AND BIOSYNTHESIS 91
..................................
formation of 34 more ATP molecules and 2 GTP mole- Under what conditions will an endergonic reaction
cules. Biological systems therefore attain efficienciesof at proceed?
least 42%, considerably better than those of any manufac- What is AG for a system at equilibrium? i
tured engine energized by the oxidation of organic fuels. How does ATP "donate" stored chemical energy to
an endergonic reaction?
What is meant by the term coupled reaction?
REVIEW QUESTIONS How does increased temperature increase the rate of
What evidence is there that the molecular building a chemical reaction?
blocks of life might have arisen spontaneously on the What factors can influence the temperature optimal
primordial Earth? for an enzymatic reaction?
What determines the reactivity of a given atom? Why? How does a catalyst increase the rate of a reaction?
What properties of carbon, hydrogen, oxygen, and Why is catalysis necessary in living organisms?
nitrogen make them especially well adapted for the How do enzymes exhibit substrate or bond specificity?
construction of biological molecules? How does pH affect the activity of an enzyme?
Why is oxygen of such biological importance? How was the "steric-fit" theory of active-site speci-
What important physical and chemical characteris- ficity shown to be correct?
tics of H,O can be directly related to the dipole na- What factors can influence the rate of enzyme-
ture of the water molecule? catalyzed reactions?
What is the pH of a 1 M solution of an acid that is The Michaelis-Menten constant, KM, is equal to the
10% dissociated? substrate concentration at which a particular reac-
Why is a weak acid rather than a strong one required tion proceeds at half its maximum velocity, V,,,,,.
for a pH buffer system? Does a high K , indicate a greater or a lesser enzyme-
What is the difference between molality and molarity? substrate affinity?
How many grams does a mole of C 0 2weigh? Why does a high substrate concentration reverse the
Approximately how many particles are in a 1M so- effects of a competitive inhibitor and yet have no ef-
lution of NaCl? fect on a noncompetitive inhibitor?
What is the approximate boiling point of a 1molal How does each type of inhibition affect the Michaelis-
solution of NaCI? Menten constant, K , ? Explain why.
Why do some liquids conduct electricity whereas oth- Why does aerobic metabolism yield much more en-
ers do not? ergy per glucose molecule than anaerobic metabolism?
How many ions flow past a point (equivalents per What is the advantage of incremental drops in elec-
second) at a current of 1mA? tron pressure compared with a single large drop in
What are the primary factors that govern the binding electron pressure in the electron-transport chain?
of two cations, a and b, to an electronegative binding How is energy liberated in discrete amounts in the
site?Write the expression that integrates these factors electron-transport chain?
into a meaningful quantity. How does the mechanism of energy release by the cit-
Does the force of attraction fall off most rapidly with ric acid cycle differ from that during glycolysis?
distance between a monovalent cation and (a) a
monopolar binding site or (b)a multipolar site? Give
SUGGESTED READINGS
the expression relating force and distance for each site.
What factors determine each level of protein struc- Atkins, P. W. 1994. Physical Chemistry. New York :W. H.
ture-primary, secondary, tertiary, and quaternary Freeman and Company. ( A complete treatment at the
protein structure? undergraduate level of many of the basic concepts in-
What special characteristic does cysteine have that troduced in this chapter.)
makes it a likely participant in the active sites of en- Lehninger, A. L., et al. 1993. Principles of Biochemistry. 2d
zyme molecules? ed. New York: Worth. (A short, straightforward book
Why do proteins become denatured (structurally dis- about biochemical principles.)
organized) at elevated temperatures? Lodish, H. D., et al. 1995. Molecular Cell Biology. 3d ed.
Living systems would appear to defy the second law of New York: Scientific American Books. (Comprehensive
thermodynamics because of their high degree of main- textbook describing many of the basic biochemical
tained order. How do you reconcile the low entropy of processes that occur in the cell.)
an organism with this fundamental physical law? Stryer, L. 1995. Biochemistry. 4th ed. New York: W. H.
At a particular temperature, will a reaction with Freeman and Co. (Highly readable reference for infor-
AS > AH be endergonic or exergonic? mation about biochemical structures and mechanism.)
T he complex chemical reactions that ultimately are re-
sponsible for animal life will proceed only under sta-
ble, restricted conditions. Such constancy is maintained
process, their existence was questioned up until the 1930s.
There was little or no direct anatomical evidence for bio-
logical membranes at the time, so their existence could only
within cells largely through the action of biological mem- be inferred from physiological studies. The first important
branes, which form a protective barrier that allows only observations on the diffusion-limiting properties of the cell
certain materials to pass into or out of the cell. Animal tis- surface were made in the mid-nineteenth century by Karl
sue contains an astounding amount of biological mem- Wilhelm von Nageli, who noticed that the cell surface acted
brane. The chimpanzee brain, for example, is estimated to as a barrier to free diffusion of dyes into the cell from the
have about 100,000 m2 of cell membrane, an area equal to extracellular fluid. From these experiments he deduced the
three full-sized soccer fields. Though cell membranes are a presence of a "plasma membrane." He also discovered the
major constituent of all living matter and essential to all life osmotic behavior of cells, noting that they swell when
placed in dilute solutions and shrink in concentrated solu-
tions. Structural evidence for the existence of a distinct cell
membrane was first available after the development of elec-
tron microscopy (see Chapter 2). At the surface of every cell
type is a continuous double-layered membrane ranging in
thickness from 6 to 23 nm (Figure 4-1).
Understanding membrane structure and function is
critical to the study of animal physiology. In this chapter we
discuss membrane structural features and their critical role
in maintaining cell integrity and controlling cell activities.
In the next chapter, we discuss the electrical behavior of cell
membranes that is responsible for cell-to-cell signaling,
which, in turn, coordinates action in animals.
MEMBRANE STRUCTURE
AND ORGANIZATION
At their surfaces, cells are surrounded by a plasma mem-
- 10 nm
brane, an extraordinarily thin, complex, lipid-based struc-
ture that encloses the cytoplasm (including the cytosol and
all cell organelles) and the cell nucleus. he internal or-
ganelles, such as the An-producing mitochondria, which
Figure 4-1 The plasma membrane creates a barrier between the interior We ~ ~ S C U in S SChapter
~ ~ 3, have their own surface mem-
and exterior of the cell, as revealed in this electron micrograph. The cell branes.) This enclosing feature of the plasma membrane is
interior (lower right) is separated from the cell exterior by the surface its most obvious function, and also its most critical. With
membrane bilayer, which is seen in cross-sectionas a dark-light profile
the help of various metabolic mechanisms, described latel;
about 10 nm thick. The dark-light-dark sandwich-likeappearance is due
to the differential staining of the "unit membrane" by an electron-
the membrane regulates molecular traffic between the or-
opaque substance during preparation of the tissue. [Courtesy of J. D. derly interior ofthe cell and themore disorderly, potentially
Robertson.] disruptive external environment.
94 PRINCIPLES O F P H Y S I O L O G Y
........................................
Membrane Composition lar head groups formed hydrogen bonds with the water and
The cell membrane is spanned by integral proteins. These their hydrophobic hydrocarbon chains stuck up into the air.
proteins act as both selective filters and active transport de- When the dispersed film of lipid molecules was gently com-
vices responsible for getting nutrients into and cellular pressed into a continuous monomolecular film, it occupied
products and waste out of the cell. Other proteins con- an area about twice the surface area of the original red
tained within the membrane sense external signals that di- blood cells. Since the only membrane in mammalian red
rect the cell responses to environmental changes. blood cells is the plasma membrane, it was concluded that
Cell membranes sustain different concentrations of cer- the lipid molecules in the membrane must be a continuous
tain ions on their two sides, leading to concentration bilayer. As we saw in Figure 4-1, the bilayer has since been
gradients of several ionic species across membranes. The visualized in cross-section by the electron microscope, as
channel proteins contained in cell membranes actively par- well as with freeze fracture methods, in which the mem-
ticipate in the translocation of substances between com- brane is split through the center of the bilayer (see Chapter
partments and ultimately regulate the cytoplasmic concen- 2). The chemical properties of lipid molecules, which cause
tration of dissolved ions and other molecules rather them to assemble spontaneously into bilayers even under ar-
precisely. This allows maintenance of an intracellular mi- tificial conditions (see Chapter 3), are responsible for the
lieu required for the finely balanced metabolic and synthetic structure of the membrane.
chemical reactions of the cell. Membranes are remarkably fluid structures, in which
All biological membranes, including the internal mem- most of the lipid and protein molecules "float" around in
branes of organelles of eukaryotic cells, have essentially the the plane of the bilayer (Figure 4-2). The relative propor-
same structure: lipid and protein molecules kept together by tion of lipids and proteins present in a membrane depends
noncovalent interactions. The lipid molecules are arranged on the kind of cell or organelle the membrane encloses.
in a continuous double layer, called the lipid bilayer, which Lipids, which are far smaller and simpler molecules than
is relatively impermeable to passage of most water-soluble proteins, provide the primary structure of the membrane.
molecules. In 1925, using a simple but elegant experiment, Integral proteins embedded in the membrane play more
Gorter and Grendel provided the first evidence that cell specialized roles such as transporting molecules through the
membranes are lipid bilayers. First, they dissolved the lipids membrane, catalyzing reactions, and transducing chemical
from red blood cell ghosts, the empty membrane sacs left signals. Other proteins connect the membrane to the cy-
when red blood cells have been induced to burst open. The toskeleton or to adjacent cells. Some proteins are intimately
extracted membrane lipids were then allowed to spread out associated with lipid molecules because of lipophilic groups
on the surface of water in a trough. Because of their asym- exposed on the surface of the protein molecule. The pro-
metry, the lipid molecules became oriented so that their po- tein-lipid complex is called a lipoprotein.
Figure 4-2 The Singer-Nicolson fluid mosaic model of the membrane oligosaccharide side chains and are vital for cell recognition and com-
is widely accepted. The globular integral proteins embedded in the lipid munication. Cholesterol molecules lie close to the heads of the phos-
bilayer provide a mechanism fortransmembrane transport The inner mi- pholip~dmolecules, where they reduce membrane flexibility. The inner
tochondrial membrane would have an even higher protein content and ends of the phospholipid tails are highly mobile, giving the membrane
thus less lipid bilayer than this figure represents. The glycoproteins bear fluidity.
MEMBRANES, CHANNELS, A N D TRANSPORT 95
Lipid molecules are insoluble in water but can be dis- and hydrophobic tails, is crucial to the organization of bi-
solved in organic solvents. They comprise about half the ological membranes. Their polar heads seek water and their
mass of plasma membranes in animal cells, the rest being nonpolar tails seek one another (see Figure 3-14), being
essentially protein. Each square micron of membrane has mutually attracted by van der Waals forces. Thus, these
about 106lipid molecules, meaning typical small cells have molecules are ideally suited to form an interface between
about lo9lipid molecules. The three primary types of lipids a nonaqueous lipid environment (phase)within the mem-
in cell membranes are brane itself and the aqueous intra- and extracellular phases
It in contact with the inner and outer membrane surfaces.
Phosphoglycerides,characterized by a glycerol backbone These same forces cause lipid bilayers to reseal themselves
when they are torn, which gives cells a self-repair capabil-
Sphingolipids, which have backbones made of sphin-
ity. Differences in the length of the two fatty acid chains and
gosine bases
in their composition (see Figure 3-20) influence lipid pack-
* Sterols, such as cholesterol, which are nonpolar and ing and hence fluidity, causing subtle differences in lipid bi-
only slightly soluble in water layer characteristics. The hydrophobic properties of the
phospholipid hydrocarbon tails are responsible for the low
If The first two lipid types are amphipathic, meaning that permeability of membranes to polar substances (e.g., inor-
they have a hydrophilic (water-soluble)and a hydrophobic ganic ions and polar nonelectrolytes such as sucrose and in-
(water-insoluble)end (Figure4-3). The dual nature of these ulin) and for their correspondingly greater permeability to
It amphipathic membrane lipids, with their hydrophilic heads nonpolar substances (e.g., steroid hormones).
Nonpolar
tails
96 PRINCIPLES OF PHYSIOLOGY
.......................................
The third class of membrane lipids, the sterols, are
largely nonpolar and only slightly soluble in water (Figure
4-4). In aqueous solution they form complexes with pro-
teins that are far more water-soluble than the sterols are
alone. Once in the membrane, the sterol molecule fits Nonpolar hvdrocarbon
talk of p h ~ s p h o ~ ~ p ~ d s
snugly between the hydrocarbon tails of the phospholipids
and sphingoliplds
and glycolipids (Figure 4-5) and increases the viscosity of
I
the hydrocarbon core of the membrane.
Integral
proteins
CH3 - + +
I
HC-CH,
I
CH2
I
CH2
Chotesterol
I
HC-CH,
Figure 4-6 A cross-sectional view shows the complexity of the mosaic bi-
layer model. The charged hydrophilic amino acid side groups of the pro-
Figure 4-4 Cholesterol, a sterol, IS an Important component of the llpld teins project into the aqueous phase, and the uncharged hydrophobic
membrane [From Lehnlnger, 19751 groups are buried in the lipid phase of the bilayer.
MEMBRANES, C H A N N E L S , AND TRANSPORT 97
................................ ......
side of the membrane to the other {ahout once a month), Hetuogenei9 of ffieintegral membrane proteins
but exchange places with adjacent molecules in a mono- The integral proteins found in the plasma membrane (see
layer about 10- times per second. This brisk lipid exchange Figure 4-2)take manv functional forms, including the ion
within a membrane results in rapid migration along the channels, various carriers and membrane pumps, receptor
plane of the membrane hut not across it. molecules, and recognition molecules. The number of inte-
Memhcane fluidly depends on its composition, and gral proteins varies, but in some membranes the protein
cholesterol plavs an important role in governing this rnem- content is so high that only about three lipid molecules sep-
brane charactezist~c.Plasma membranes in eukarvotic or- arate the proteins a t the point of closest approach.
ganisms contain lots of cholesterol, up to one molecule for .Morphological evidence for the hererogeneous mosaic
every phospholipid. Cholesterol, when present, binds arrangement of globular proteins in a lipid bilaper is seen
weakly to adjacent phospholipids, making lipid hilapers in freeze-etch electron micrographs of the surface of a
significantly less fluid, but stronger (Figure 4-7). The incor- membrane (Figure 4-8). When subjected to digestion by
poration of too much cholesterol into cell membranes,
however, causes the membranes to lose flexibility. This ir
the mechanism underlying "hardening of the arteries," a
major cause of cardiovascular disease, in which the ceEl
membranes of the endochelia! cells lining the arteries be-
come abnormally rigid (with additional cholesterol plaques
stored in the intirnal lavers, as well).
Lipid composition of biological membranes varies
among tissue tvpes. While most membranes contain a sig-
nificant fraction of cholesterol ( > I S % ) , other types of
lipids may he present in much higher or lower fractions.
Lipids also differ in their head groups (see Figure 3-1 3),
which in turn influences their interactions with proteins. In-
deed, some integral proteins function only in the presence
of a specific ratio of lipid types. Hence, cells must regulate
the distribution of lipid species in their membranes during
cell development and rearrange lipid concentrations ac-
cording to specific functional needs.
Phospholipid
Cholesterol
proteolytic (protein-digesting) enzymes, the globular units drophobic interior of the lipid bilayer makes membranes
seen in the membrane are progressively removed, demon- highly impermeable to most polar molecules. This prevents
strating that they are indeed proteins. water-soluble components of the cell from easily entering
or escaping. However, such movement may at times be nec-
Variation in Membrane Form essary or desirable, so mechanisms for transferring these
Membrane composition varies greatly between cell types. molecules across membranes have evolved in all cells.
At one extreme is the metabolically inert myelin sheath sur- Macromolecules like proteins and large particles must also
rounding the axons of some nerve cells, in which the lipid be transported across plasma membranes using specialized
bilayer is largely uninterrupted. At the other extreme are mechanisms.
cells with membranes that have structures of repeating non- To understand these special means of membrane trans-
lipid macromolecular units that nearly obliterate the lipid port in living cells, we will first review the physical princi-
bilayers. Such membranes evolved for highly specialized ples of solute and solvent displacement in solution and
purposes such as signaling or enzymatic activity. In visual across semipermeable membranes. Such membranes closely
receptor cells, for example, the repeating macromolecular resemble those found in living cells, and the principles ex-
unlts are molecules of the visual pigment opsin. Mitochon- plained here apply in many physiological situations.
drial membranes, specialized for enzymatic activity, are
composed almost entirely of repeating subunits of ordered Diffusion
enzymatic aggregates. Between these extremes are the Random thermal motion of suspended or dissolved mole-
plasma membrane and most intracellular membranes, in cules causes their dispersion from regions of higher con-
which the bilayer is interrupted frequently by integral pro- centration to regions of lower concentration, a process
tein molecules. Thus the basic structure of the lipid bilayer called diffusion. Diffusion is extremely slow when viewed
with integral proteins is highly modified as required for on a tissue, rather than a cellular, scale. For example, a
functional specialization. crystal of copper sulfate dissolves in unstirred water so
slowly that it may take a whole day to color a liter of water
completely. When viewed in the microscopic dimensions of
CROSSING THE MEMBRANE:
the cell, however, diffusion times can be as short as a frac-
AN OVERVIEW
tion of a millisecond.
The structure of membranes makes them quite selective The rate of diffusion of a solute s can be defined by the
about which molecules can pass through them. The hy- Fick diffusion equation:
MEMBRANES, C H A N N E L S , A N D TRANSPORT 99
.............................................................................
Semipermeable
Membrane Flux
If a solute occurs on both sides of a membrane through
which it can diffuse, it will exhibit a unidirectional flux in
each direction (Figure 4-9A). The flux, or rate of diffusion,
Jis the amount of the solute that passes through a unit area
of membrane every second in one direction, so that
Net flux
where J would typically have units of moles per square cen-
Figure 4-9 Solutes can move through a membrane in either dired~on,
timeter per second (M cm-2. s-l). The flux in one direc-
depending on prevailing physical and chemical conditions. (A) The ar-
tion (say,from cell exterior to cell interior) is considered in- rows represent the actual fluxes of a substance between compartments I
dependent of the flux in the opposite direction. Thus, if the and II. (B) The single arrow indicates the resulting net flux, from com-
influx and efflux are equal, the net flux is zero. If the uni- partment Ito II.
directional flux is greater in one direction, there is a net flux,
which is the difference between the two unidirectional
fluxes (Figure 4-9B).
The permeability of the membrane to a substance is the
rate at which that substance passively penetrates the mem- concentration gradient, but also on the electrical gradient
brane under a specified set of conditions. A greater perme- (i.e., the electric potential difference across the membrane).
ability will be accompanied by a greater flux if other factors As is evident from equation 4-3, the flux of a nonelectrolyte
remain equal. If we assume that the membrane is a homo- should be a linear function of the concentration gradient
geneous barrier and that a continuous concentration gra- (C, - C,). This linear relationship is characteristic of sim-
dient exists for a nonelectrolyte substance between the side ple diffusion, and can be used in experiments to distinguish
of high concentration (I)and the side of low concentration between passive diffusion of a substance and any other
(II),then mechanism. The permeability constant incorporates all the
factors inherent in the membrane and the substance in
dQs
- - - PIC, - C,)
question. These factors will determine the probability that
dt a molecule of a particular substance will cross the mem-
brane. This relationship can be expressed formally as
in which dQ,ldt is, again, the amount of substance s cross-
ing a unit area of membrane per unit time (e.g., moles per
square centimeter per second), C, and C,, are the respective
.
concentrations (e.g., M ~ m - of~ the
) substance on the two
sides of the membrane, and P is the permeability constant where Dmis the diffusion coefficient of the substance within
of the substance, with the dimension of velocity (cm-s-I). the membrane, K is the partition coefficient of the sub-
Note that equation 4-3 applies only to molecules that stance, and x is the thickness of the membrane. The more
are not being actively transported or influenced by any viscous the membrane or the larger the molecule, the lower
forces other than simple diffusion. This excludes elec- the value of Dm.
trolytes, since they are electrically charged when dissoci- Permeability constants for different substances vary
ated, and consequently their flux depends not only on the greatly. For example, the permeability of red blood
100 PRINCIPLES OF PHYSIOLOGY
..........................................
cells to different solutes ranges from 10-12 cm-s-I to producing membranes that would allow water molecules
-
cm s-l. Importantly, the permeability of some mem- to diffuse through them far more freely than sucrose mol-
branes to certain substances can be altered greatly by hor- ecules could. These artificial membranes were also strong
mones and other molecules that react with receptor sites on enough to withstand relatively high pressures without rup-
the membrane and thereby influence channel size or carrier turing because of the clay substratum. Using these mem-
mechanisms. Antidiuretic hormone, for example, can in- branes, Pfeller was able to make the first direct measure-
crease the water permeability of the renal collecting duct in ments of osmotic pressure. Some of his results are shown in
mammals by as much as 10 times. Similarly, neurotrans- Table 4-1. Note in the table that the osmotic pressure is
mitters, acting on specialized integral membrane proteins proportional to the solute concentration.
in nerve and muscle cells, induce large increases in perme- Osmosis is responsible for the net movement of water
ability to ions such as Nac, KC, Ca2+,or C1-. across cell membranes and epithelia. To understand this,
consider a 1.0 M aqueous solution of sucrose carefully lay-
Osmosis ered under a 0.01 M aqueous solution of sucrose. There
In 1748 AbbC Jean Antoine Nollet noted that if pure wa- would be net diffusion of water molecules from the solu-
ter is placed on one side of an animal membrane (e.g., a tion of lower sucrose concentration (the 0.01 M solution)
bladder wall) and a solution of water containing elec- into the 1.0 M sucrose solution, and sucrose would show
trolytes or other molecules is placed on the other side, the net diffusion in the opposite direction until equilibrium was
water passes through the membrane into the solution. This achieved. If these two solutions were separated by a mem-
movement of water down its concentration gradient was brane permeable to water but not to sucrose, the water
called osmosis (from the Greek osmos, "to push"). We molecules would still show a net diffusion from the solu-
don't usually think of "water concentration," but in fact tion in which H,O is more concentrated (the 0.01 M su-
water acts just like any other substance by diffusing down crose solution) into the 1.0 M sucrose solution, in which
its concentration gradient. It was later found that osmosis the H 2 0 concentration is lower. Since the sucrose could not
produces a hydrostatic pressure gradient. Osmosis is the cross the membrane, there would be a net diffusion of wa-
colligative property of greatest importance to living sys- ter (osmoticflow) through the membrane from the solution
tems. As can be seen in Figure 4-10, the pressure difference of lower solute concentration to the solution of higher
causes a rise in the level of the solution as water diffuses solute concentration.
through the semipermeable membrane into the solution. Osmotic pressure .iris proportional not only to the con-
The rise in the level of the solution continues until the net centration of the solute, C (moles of solute particles per liter
rate of water movement (net flux) across the membrane be-
comes zero. This occurs when the hydrostatic pressure of TABLE 4-1
the solution in compartment I1 is sufficient to force water Osmotic pressure of sucrose solutions of various
molecules back through the membrane to compartment I concentrations*
at the same rate that osmosis causes water molecules to dif- Sucrose Osmotic pressure Ratio of osmotic pressure
fuse from I to 11. The hydrostatic back pressure required to (%I (atm) t o percentage of sucrose
cancel the osmotic diffusion of water from compartment I
1 070 070
to compartment I1 is called the osmotic pressure of the so-
2 1.34 067
lution in compartment 11.
In 1877, Wilhelm Pfeller made the first quantitative 4 2 74 068
studies of osmotic pressure. He deposited a "membrane" 6 4.10 068
of copper ferrocyanide on the surface of porous clay cups, *Results were obtarned by Pfeffer (1877) in exper~mentalmeasurements
Semipermeable
6 membrane
-
Time Figure 4-10 Water flow produced by osmosis through a semipermeable
membrane generates hydrostatic pressures. Compartment I contains
pure water; compartment II, water with impermeant solute. Osmotic
pressure forces water to enter compartment II from compartment I until
the hydrostatic pressure difference equals the opposing osmotic pres-
sure difference. When the pressures are equal, the flux is zero.
MEMBRANES, C H A N N E L S , A N D TRANSPORT 101
.......................................
of solvent = osmolarity), but also to its absolute tempera- to be hypertonic to it. These effects result from movement
ture T: of water across the cell membrane in response to osmotic
pressure differences between the cell interior and the extra-
TT = KIC (4-5) cellular solution.
If cells actually behaved as ideal osmometers, tonicity
and and osmolarity would be equivalent, but this is not gener-
ally true. For example, sea urchin eggs maintain a constant
volume in a solution of NaCl that is isosmotic relative to
seawater, but they swell if immersed in a solution of CaC1,
where K, and K, are constants of proportionality. Jacobus that is isosmotic relative to seawater. The NaCl solution
van't Hoff related these observations to the gas laws and therefore behaves isotonically relative to the sea urchin egg,
showed that solute molecules in solution behave thermo- whereas the CaCI, solution behaves hypotonically. The
dynamically like gas molecules. Thus, tonicity of a solution depends on the rate of intracellular ac-
cumulation of the solute in the tissues in question, as well
TT = RTC as on the concentration of the solution. The more readily
the solute accumulates, the lower the tonicity of a solution
of a given concentration or osmolarity. This is because as
the cell ,gradually loads up with the solute, water follows
according to osmotic principles, causing the cell to swell.
Thus, the terms isotonic, hypertonic, and hypotonic are
meaningful only in reference to actual experimental deter-
where n is the number of mole equivalents of solute, R is minations on living cells or tissues.
- .
the molar gas constant (0.082 L atm K-l mol-' )," and
Electrical Influences on Ion Distribution
V is the volume in liters. Like the gas laws, however, this ex-
pression for osmotic pressure holds true only for dilute so- Membrane permeability to charged particles depends both
lutions and for completely dissociated electrolytes. on the membrane permeability constant and on the electri-
Large concentration gradients across cell membranes cal potential across the membrane. Understanding the in-
can generate surprisingly high osmotic pressures-on the teraction of charged particles with membranes is extremely
order of several atmospheres. Such pressures, if allowed important for understanding how electrically excitable cells
to develop, would be large enough literally to explode function. Neurons are the most highly specialized of this
a cell. Consequently, mechanisms for regulating osmotic class of cells. Since neurons will be discussed in the next
balance have evolved that minimize osmotic pressure gra- couple of chapters, only a few important observations will
dients across cell membranes and through tissues (see be summarized here.
Chapter 14). Two forces can act on charged atoms and molecules
(such as Na+,K+, Clk, Ca2+,amino acids) to produce a net
Osmolarity and Tonicity passive diffusion of each species across a membrane:
Two solutions that exert the same osmotic pressure
through a membrane permeable only to water are said to 1. The chemical gradient arising from differences in the
be isosmotic to each other. If one solution exerts less os- concentration of the substance on the two sides of the
motic pressure than the other, it is hypoosmotic with re- membrane
spect to the other solution; if it exerts greater osmotic pres- 2. The electric field, or difference in electric potential
sure, it is hyperosmotic. Osmolarity is thus defined on the across the membrane
basis of an ideal osmometer in which the osmotic mem-
brane allows water to pass but completely prevents the An ion will move away from regions of high concentration,
solute from passing. All solutions with the same number of and if that ion is positively charged it will also move toward
dissolved particles per unit volume have the same osmolar- increasing negative potential. The sum of the combined
ity'and are thus defined as isosmotic. forces of concentration gradient and electrical gradient de-
The tonicity of a solution, in contrast to its osmolarity, termine the net electrochemical gradient acting on that ion.
is defined by the response of cells or tissues immersed in the When an ion is at equilibrium with respect to a mem-
solution. A solution is considered to be isotonic with a brane (that is, when there is no net transmembrane flux of
given cell or tissue if the cell or tissue immersed in it neither that ion species), there will exist a potential difference just
shrinks nor swells. If the tissue swells, the solution is said to sufficient to balance and counteract the chemical gradient
be hypotonic to the tissue; if it shrinks, the solution is said acting on the ion. The potential at which an ion is in elec-
trochemical equilibrium is called the equilibrium potential,
measured in volts (or millivolts). Several factors influence
'R is the constant of proportionality in the gas equation PV/T = R
the value of the equilibrium potential, but the most promi-
when referring to 1 mol of a perfect gas, and it has the value of
1.985 cal. molk' .K-'; P is in atmospheres and V is in liters. nent is the ratio of the ion concentrations on opposite sides
102 PRINCIPLES O F PHYSIOLOGY
.......................................
of the membrane. For a monovalent ion such as Na+ or K+ A
Donnan Equilibrium
If diffusible solutes are separated by a membrane that
is freely permeable to water and electrolytes but totally
Figure 4-1 1 The Donnan equilibrium descr~besion distribution across a
impermeable to one species of ion, the diffusible solutes semipermeable membrane. (A) When KC1 is added to compartment I of
become unequally distributed between the two com- a container divided by a permeable membrane, K+ and CI- diffuse across
partments. This phenomenon was discovered in 1911 by the membrane until the concentrationsare equal on either side. (B) If the
Frederick Donnan, who first described how the solutes potassium salt of an impermeant anion is added to compartment I, some
K+ and C I diffuse into compartment II until electrochemicalequilibrium
would be distributed and hence has been commemorated
is reestablished. It should be noted that these chambers (unlikethe living
by having this equilibrium state named for him. cell) are not distensible.
To understand a Donnan equilibrium, imagine starting
with pure water in two compartments and adding some
KC1 to one of them (Figure 4-11). The dissolved salt (K+
and C1-) will diffuse through the membrane until the sys-
We can understand this situation by considering the
tem is in equilibrium-that is, until the concentrations of
consequences of the following physical principles:
K+ and C1- become equal on both sides of the membrane
(Figure 4-11A). Now imagine adding the potassium salt of
a nondiffusible anion (a macromolecule A-, having multi- 1. There must be electrone~traiit~
within both compart-
ments; that is, in each compartment the total number
ple negative charges)to the solution in compartment I. The
K+ and C1- quickly become redistributed until a new equi- of positive charges must equal the total number of neg-
ative charges. Thus, in this example, [K+] = [Cl-] in
librium is established by movement of some K+ and some
compartment 11.
C1- from compartment I to compartment I1 (Figure4-11B).
2. Considered statistically, the diffusible ions K+ and C1-
Donnan equilibrium is characterized by a reciprocal distri-
cross the membrane in pairs to maintain electrical neu-
bution of the anions and cations so that
trality. The probability that they will cross together is
proportional to the product [K+]X [Cl-1.
3. At equilibrium the rate of diffusion of KC1 in one di- '
rection through the membrane must equal the rate of
KC1 diffusion in the opposite direction. It follows,
At equilibrium, the diffusible cation, K+, is more concen- then, that at equilibrium the product [K+] x [Cl-] in
trated in the compartment in which the nondiffusible an- one compartment must equal the product in the other
ion, A-, is confined than in the other, whereas the diffusible compartment. Letting x , y, and z represent the con-
anion, C1-, becomes less concentrated in that compartment centrations of the ions in compartments I and 11, as
than in the other. shown in Figure 4-12, we can express the equilibrium
meable to a variety of ions and molecules, and there will al-
most never be a single "nondiffusible anion," which here
represents various anionic side groups of proteins and other
large molecules. Although the physical and mathematical
principles recognized by Donnan play a role in regulating
the distribution of electrolytes in living cells, clearly non-
equilibrium mechanisms must modify the distribution of
many substances across the cell membrane. In particular,
Figure 4-12 The Donnan equilibrium can be described algebraically. the permeability of the cell membrane to particular ions can
The equilibrium condition established in Figure 4-1 1 B afterthe salt of an
impermeant anion is added to compartment 1 is shown.
change over time, changing the conditions dramatically.
Thus, cells cannot be considered passive "osmometers,"
and the distribution of substances across biological mem-
branes cannot be predicted entirely by Donnan equilibrium
principles except in certain cases.
condition (i.e., equality of the product [K+] x [Cl-] in
the two compartments) algebraically:
OSMOTIC PROPERTIES OF CELLS
We can now use the physical principles outlined above to
analyze properties of the cell membrane that maintain dif-
This equation also holds, of course, if A- is not present. In ferent concentrations of ions inside and outside the cell
that case, K+ and C1- are equally distributed, and z = 0 (Figure 4-13). Cell membranes ultimately must closely reg-
and x = y. By rearranging equation 4-8, we can see that, at ulate cell volume and thus intracellular osmotic pressure.
equilibrium, the distributions of the diffusible ions in the
two compartments are reciprocal: Ionic Steady State
Every cell maintains concentrations of inorganic solutes in-
side the cell that are different from those outside the cell
(Table 4-2).The most concentrated inorganic ion in the cy-
tosol is K+, which is typically 10-30 times as concentrated
From this relation, it is clear that as the concentration of the there as in the extracellular fluid. Conversely, the internal
nondiffusible anion, z, is increased, the concentrations of concentrations of free Na+ and C1- are typically less (ap-
the diffusible ions (x and y) will become increasingly diver- proximately one-tenth or less) than the external concentra-
gent. This unequal distribution of diffusible ions is the hall- tions. Another important generalization is that the intra-
mark of Donnan equilibrium. cellular concentration of Ca2+is maintained several orders
At Donnan equilibrium, the osmotically unequal distrib- of magnitude below the extracellular concentration. This
ution of solute particles makes water move in the direction of difference is due in part to active transport of Ca2+ out
the compartment of higher osmolarity (compartment I in across the cell membrane and in part to the sequestering of
Figure 4-11). This osmotic pressure difference plus any re- this ion within such organelles as the mitochondria and en-
sultant increase in hydrostatic pressure of that compartment doplasmic reticulum. As a result, the concentration of Ca2+
is called the oncotic pressure. This concept is important in un- in the cytosol is generally well below M.
derstanding the balance of hydrostatic and osmotic pressures Cell membranes typically are about 30 times more per-
across certain biological barriers such as capillary walls. meable to K+ than to Na+. Membrane permeability to
The explanation of a Donnan equilibrium depends on chloride ions varies. In some cells it is similar to that of K+
an ideal set of conditions for the sake of simplicity. The liv- while in others it is lower. The permeability of the cell mem-
ing cell and its surface membrane are, of course, far more brane to Na' is low, but it is not low enough to prevent
complex. For example, the cell membrane is somewhat per- Na+ from leaking steadily into the cell.
N o active
transport
of Na+
Figure 4-15 Osmotic changes alter the volume of a red blood cell. Cell bursts
(A) Isotonic solution: the cell volume.remainsunchanged. (B) Hypotonic
solution: water (arrows)enters the cell because of the higher osmoticity
of the cytoplasm with respect to the solution, producing swelling. (C) Hy-
pertonic solution: in a more concentrated medium, water leaves the cell,
causing shrinkage.
t Hyperosmotic solution
added
,
u Time
that while these processes do not directly require metabol-
ically energized processes, they ultimately depend on a con-
centration or electrical gradient across a cell membrane that
Figure 4-16 Hyperosmotic solutions with impermeant and weakly has at some point required energy for its creation and main-
impermeant solute both cause initial cell shrinkage. If the solute is com- tenance. The energy stored in such gradients is ultimately
pletely impermeant, it causes maintained cell shrinkage because responsible for the translocation of molecules across the
the solution is basically hypertonic in this situation. If the solute is only membrane. Nonetheless, it is useful and appropriate to
weakly impermeant, however, the solution IS hypotonic and enters
the cell slowly followed by the osmotic flow of water. This process even-
think of these processes as passive.
tually produces swelling, in spite of the fact that the solution is hyper- There are three basic routes for passive transmembrane
osmotic. movements of molecules or ions (Figure4-18).In the first, a
106 PRINCIPLES OF PHYSIOLOGY
...............................................................................
w
A leave the aqueous phase and enter the lipid phase, a solute
Outside Inside must first break all its hydrogen bonds with water. This re-
quires about 5 kcal of kinetic energy per hydrogen bond.
Moreover, the solute molecule crossing the lipid phase of
.-
"- the membrane must dissolve in the lipid bilayer. So, its lipid
*a solubility will also play a major role in determining whether
m
~II or not ~twill cross the membrane. Consequently, those mol-
ecules having a minimum of hydrogen bonding with wa-
substrate Extracellular ter will most readily enter the lipid bilayer, whereas polar
molecules
molecules such as inorganic ions will almost never dissolve
in the bilayer.
A number of factors, such as molecular weight and
molecular shape, influence the mobility of nonelectrolytes
within the membrane, but the empirically measured parti-
tion coefficient is the primary predictor of the diffusion of
a nonelectrolyte across the lipid bilayer. To measure this
property, a test substance is shaken in a closed tube con-
taining-equal
.
amounts of water and olive oil, and the coef-
C Carrier ficient K is determined from the relative solubilities in wa-
molecules ter and oil at equilibrium, using the equation
Figure 4-20 The structure of six-carbon molecules determines their wa- Number of
ter and lipid solubility. Note the difference between hexanol and manni- hydrogen bonds
to1 in the number of hydroxyl groups. Hexanol, with its weak hydrogen-
bonding capacity, is poorly soluble in water and highly soluble In lipids; Figure 4-21 Hydrogen bonds greatly decrease the l ~ p solublllty
~d and,
mannitol, with its strong hydrogen-bonding capacity, is highly soluble in hence, the permeablllty of a membrane.
water and poorly soluble in lipids.
A
Extracellular space - Channel
-
Fiqure 4-22 Membrane transport . proteins
.
act as carriers or form channels in the mern-
brane. (A) A channel protein forms a water-
Lipid filled pore across the bilayer, through which
protein
bilayer specific ions can diffuse. (B)In contrast, a car-
rier protein alternates between two confor-
Cytoplasm
mations, so that the solute bindinq site is se-
B quentially accessible on one side of the bilayer
Solute Carrier Extracellular and then on the other.
" .. .
SPOTLIGHT 4-2 The principle of bilayer formation is shown in the figure
(part B). The most stable configuration attained consists of two
ARTIFICIAL BILAYERS layers of lipid molecules whose hydrophobic, lipophilic hydro-
carbon tails are loosely associated to form a liquid-lipid phase
Many of our ideas of how molecules and ions pass across mem- sandwiched between the hydrophilic polar ends of the mole-
branes have grown out of experiments and observations on ar- cules, which are directed outward toward the aqueous medium.
tificial bilayers that are similar to the bimolecular leaflet that The thickness of the lipid film is easily determined from the in-
forms the basis of the cell membrane. Artificial bilayers are ex- terference color of light reflected from the two surfaces of the
tremely useful in studies of permeation mechanisms because film. Membranes with thicknesses of approximately 7 nm (black
they can be made from chemically defined mixtures of lipids. Se- interference color) are most commonly used. These membranes
lected substances can be added to test their effects on perme- have electrical conductances (ion permeabilities) and capaci-
ability. Channel-forming substances, such as the antibiotic tances consistent with their thickness and lipid composition. Al-
ionophores (moleculesthat facilitate the diffusion of ions across though their permeability to ions is much lower than that of cell
membranes) and membrane channel components of excitable membranes, the addition of certain ionophores increases it to
tissues, have been incorporated into artificial bilayers, allowing values that are characteristic of cell membranes.
their properties to be studied in isolation under the highly con-
trolled conditions shown in the accompanyingfigure.
Bilayer-filled
1-mm opening
Lipid bilayers can be induced to form across a I-mm opening between l i p ~ din a solvent such as hexane. Initially, while the bilayer is forming, its
two fluid-f~lledchambers. (A) The permeability of the bilayer to elec- interference color is gray (left).As the membrane assumes the more sta-
trolytes in the chamber can be measured electrically by placing test so- ble bilayer configuration (right),the interference color changes to black.
lut~ons with different electrolyte concentrations in each of the chambers. [From Kotyk and Janacek, 1970.1
(B)The bilayer is formed by filling the opening with a small amount ofthe
The functioning of membrane channels can be demon- nels that allow thousands of ions per second to stream
strated directly in artificial lipid bilayer membranes that are down their gradients and across the membrane.
by themselves highly impermeable to even the smallest of Studies of the permeabilities of cell membranes to
charged molecules (Spotlight 4-2). A dramatic increase in other polar substances give an estimated 0.7 nm for
ion permeability occurs upon addition of small amounts of the equivalent pore size-the pore diameter that would .
channel proteins extracted from cellular membranes. This account for the rate of diffusion across the membrane.
increase is measured as discrete pulses of current carried by Thus, membrane channels presumably have diameters
ions from one side of the membrane to the other, just like of less than 1.0 nm, close to the practical limits of resolu-
those measured in biological membranes. These unitary tion of contemporary electron microscopes and fixation
currents are due to the sudden opening of individual chan- methods.
As an example, rod-shaped molecules of the antibiotic
nystatin applied to both sides of an artificial or a natural
membrane aggregate to form channels. These pores permit 5
the passage of water, urea, and chloride, all of which are e,
Y
m Facllltated diffusion
less than 0.4 nm in diameter. Larger molecules cannot
penetrate the channels. Cations also are excluded, presum-
ably because there are fixed positive sites along the channel
walls. Incorporation of nystatin into artificial membranes
produces a negligible increase in membrane area occupied
by fixed channels (0.001%-0.01%), but it produces a
Passive diffusion
100,000-fold increase in membrane permeability to chlo-
ride ions. This means that very little membrane area need
be devoted to channels to account for the ion permeabili- I External concentration of glucose (mM)
ties of natural membranes. This conclusion is supported Km
by the fact that the electrical capacitance of the cell mem-
Figure 4-24 The kinetics of simple diffusion differ from those of carrier-
brane remains relatively unchanged during large changes
med~ated(facilitated) diffusion. In this example of glucose movements,
in the permeability exhibited during the excitation of the rate of simple diffusion is always proportional to the glucose con-
some membranes. (This phenomenon is discussed further centration. However, the rate of carrier-mediated glucose diffusion
in Chapter 5.) reaches a maximum (V,,,) when the glucose carrier protein is saturated.
The binding constant (K,) of the carrier for glucose, which is analogous
Facilitated Transport across Membranes to the Kmof an enzyme for its substrate solute, is measured when trans-
port is at half its maximal value. [Adapted from Lodish et al., 1995.1
Membranes are permeable to various polar molecules such
as sugars, amino acids, nucleotides, and certain cell
metabolites that would cross lipid bilayers by diffusion only
very slowly. This is because of facilitated transport, the The existence of such transporters was initially inferred
movement of molecules through membranes by the action from kinetics studies of molecule transfer across mem-
of membrane transport proteins (see Figure 4-22B). Facili- branes (Figure 4-24). For some solutes, the measured rate
tated transport, unlike active transport discussed later, does of influx reaches a plateau beyond which an increase in
not require energy in the form of ATP. Membrane transport solute concentration produces no further increase. This re-
proteins, which exist in many forms in all types of mem- veals that a rate-limiting step must occur in permeation. Ex-
branes, are exquisitely selective about which species of mol- periments elucidating the kinetics of such permeation led to
ecules they transport. Carrier proteins that transport a sin- the conclusion that transport occurs through the formation
gle solute from one side of the membrane to the other are of a carrier-substrate complex similar in concept to an en-
called uniporters, while those that transfer one solute and zyme-substrate complex. Each carrier protein has a char-
simultaneously or sequentially transfer a second solute are acteristic binding constant for its solute equal to the con-
called coupled transporters. Coupled transporters that centration of solute when the transport rate is half its
transfer two solutes in the same direction are called sym- maximum value (see Figure 4-24). As in enzyme reactions,
porters, while those that transfer solutes in opposite direc- the solute binding can be blocked by specific competitive in-
tions are called antiporters (Figure 4-23). These terms can hibitors as well as by noncompetitive inhibitors. The car-
also be applied to active transport systems. rier and solute molecule temporarily form a complex based
on bonding, stearic specificity, or both.
7 i Extracellular
The specificity of these transporters was first estab-
lished in studies where single gene mutations abolished
the ability of bacteria to transport specific sugars across
their cell membranes. Similar mutations have now been
found in many cases, including human inherited diseases
that affect the transport of specific solutes across kidney,
intestine, or lungs. For example, in cystic fibrosis, a de-
fect in the chloride transport channel protein (CFTR)
appears to be responsible for fluid imbalance in the
Uniporter Symporter Antiporter lungs.
0
Potassium
electrochemical
molecule of ATP hydrolyzed directly to power transmembranetransport,
three Na+ ions are pumped out and two K+ ions are pumped in. The spe-
cific pump inhibitor ouabain and K+ compete for the same sites on the
external side of the ATPase. (B)This schematic model of Na+/K+ATPase
gradient shows the movement of Na+ and K+ by a single protein. The binding of
1 Na+ (step 1) and the subsequent phosphorylationof the cytoplasmicface
ofthe ATPase by ATP (step 2) cause a conformational change in the pro-
ATP A D P + Pi tein resulting in a transfer of the Na+ across the membrane (step 3). The
Na+ is releasedto rhe cell's exterior and K+ is bound (step 4). Subsequent
dephosphorylationofthe protein (step 5) induces a return to the protein's
original conformation and consequent transfer of the Kt across the mem-
brane (step 6), where it is released into the cytosol (step 7).
Na+blndlng K+ b~ndlng
sites sltes
ADP + Pi
A G O ' =
- ATP + H,O
+7.3 kcal-mol-I
Chemiosmotic energy transduction similar to that pro-
posed for oxidative phosphorylation in mitochondria has
been implicated as the mechanism for energy transduction
during photosynthesis in chloroplasts and photosynthetic
bacteria. In addition, there is evidence that the Na+/K+
This reaction also requires that an ATPase complex be pump, which normally utilizes ATP to produce the Na+
oriented on the inner mitochondria1 membrane so as to gradient, can in special circumstances run in reverse, so that
take advantage of the separation of H+ and O H across the the movement of Na+ down its gradient will cause the
membrane. The H+ that is enzymatically removed from pump to synthesize ATP from ADP and P, .
Inner
mitochondrial
membrane
H+
lntermembrane Mitochondria1
space
H+
II OH-
matrix
Time
HOH
H+
\
Carrier protein
'1 Cytosol
Figure 4-29 Sugar and amino a c ~ dtransport may be ach~evedby port IS Inward because ofthe Na+ gradlent, Indicated by the arrow Note
sodium-medlated cotransport The carrier proteln must b ~ n d both the that glucose IS movlng agatnst ~tsgradlent
Na+ and the organlc substrate before it will transport elther. Net trans-
used to drive several energy-requiring processes in the arises from the Na+ concentration gradient, directed from
membrane. the lumen into the cell. This gradient is maintained by the
removal of Na+ from the cell by the Na+/K+pump located
Antiporters in the membrane on the other side of the cell, which faces
The Na+ concentration gradient also plays a role in the the plasma and blood.
maintenance of a very low intracellular Ca2+concentration
in certain cells via the Na+/Ca2+antiport system. In most,
if not all cells, the intracellular Ca2+concentration is sev-
MEMBRANE SELECTIVITY
eral orders of magnitude below the extracellular concen- The utility of cell membranes lies in their selectivity-their
trations (less than M), and many cell functions are ability to allow the passage of only specific types of mole-
regulated by changes in the intracellular Ca2+concentra- cules. This selectivity is important because a nonselective
tion. Efflux of Ca2+from cells is reduced when extracellu- membrane will not protect the contents of the cell from in-
lar Na+ is removed, because Ca2+is expelled from the cell trusion by unwanted chemicals. Each kind of membrane
in exchange for Na+ leaking in. The opposing movements transport system also displays selectivity, which differs in
of these two ions are coupled to each other by an anti- a given membrane for different transport systems. For ex-
porter. One view is that Ca2+and Na+ both compete for ample, when the Na+ in a physiological saline solution used
the carrier, but that Ca2+competes more successfully inside to bathe a nerve cell is replaced with lithium ions, the Li+
the cell than on the outer surface, so that there is a net ef- readily passes through the Na+ channels, which open dur-
flux of Ca2+.Here, again, the immediate source of energy ing electrical excitation of the nerve cell membrane. The
is the Na+ gradient, which ultimately depends on the ATP- other alkali metal cations, K+, Rb+, and Cs+, are essentially
energized active transport of Na+. Ca2+is also transported impermeant through these channels. On the other hand,
independently of the Na+ gradient by an ATP-energized the ATPase of the Na+ pump in the same membrane is
Ca2+pump, which is the major source of Ca2+extrusion highly specific for intracellular Na+ and is not activated by
under normal conditions. Li+. Lithium ions, passing through the Na+ channels, will
The Na+/H+ antiporter in the proximal tubule of the therefore gradually accumulate in the cell until it comes
mammalian kidney is another example of cotransport in into electrochemical equilibrium. This is an example of
opposite directions (see Chapter 14).Here the extrusion of electrolyte selectivity by the transport system, but not by
H+ from inside the cells lining the renal tubule into the the membrane channels. We will now consider how this
urine contained within the tubule is coupled to Na+ uptake selectivity for both electrolytes and nonelectrolytes is
into the cell in a 1:1stoichiometry. That is, for each H+ ex- achieved.
pelled, one Na+ is taken up into the cell. This has the ad-
vantages of (1)avoiding the expenditure of energy to per- Selectivity for Electrolytes
form electrical work, since two equivalent positive charges How do channels discriminate between different ions? Al-
are exchanged and (2)enabling the kidney to reclaim Na+ though enzymes recognize substrates via distinct shapes or
from the urine and excrete excess protons. The Na+/H+ex- chemical structures, membranes can distinguish ions of es-
changer, unlike the Na+/K+pump, is oriented so as to move sentially identical shape and size. For example, Na+ and K+
Na+ out of the lumen and into the cell. Also unlike the have almost the same shape and size (K+is a little larger),
Na+/K+ pump, this mechanism is not an example of pri- yet the resting nerve cell membrane is about 30 times more
mary active transport, in which ATP is the immediate permeable to K+ than to Na+. At first glance, we might
source of energy. Instead, the Na+/H+exchanger is an ex- conclude that these ions are distinguished on the basis of
ample of secondary active transport, in which the source of their hydrated size, with K+ passing freely through chan-
energy is the electrochemical gradient of one or both ex- nels that are too small for Na+. Size can explain how the
changed ions. In this case the energy driving the exchange K+ channel excludes Cs+ or Rb+ (Table 4-3) but not Na+,
......................................
TABLE 4-3 Channel
Ionic radii and hydration energies of the alkali metal cations.
Free energy of hydrat~on
Cat~on l o n ~ crad~us(A) .
(kcal mol-"
Ll + OM) -122
Na+ 0 95 -98
K+ 1 33 - 80
Rb+ 1 48 -75
CS+ 1.69 -67
Lipid bila
A Plasma membrane 6
Figure 4-33 Gap junctions permit passage of molecules between neigh- the connected cells. (B) Detail of a channel complex. Molecules smaller
boring cells. (A)The two membranes belonging to neighboring coupled than about 2 nm can pass between coupled cells through the channel.
cells both contain an array of hexagonal subunits, each of which connects Molecules larger than 2 nm, such as proteins and nucleic acids, are too
with a matching subunit in the apposed membrane. A central channel large to penetrate the channel. [Part Aadapted from Staehelin, 1974; part
penetrates both subunits, providing a path of communication between B adapted from Bretscher, 1985.1
amino acids, sugars, and nucleotides, are easily exchanged into a coupled cell, by lowering temperature, or by using
between cells (see Figure 4-33B). This exchange of small poisons that inhibit energy metabolism. The subsequent
molecules is responsible for gap junction-mediated cell-cell loss of electrical transmission between cells confirms the
communication. uncoupling. Thus, gap junctions are maintained intact only
Gap junctions are labile and close rapidly (within sec- if the metabolic activity of the surface membrane maintains
onds) in response to any treatment that increases intracel- sufficiently low concentrations of intracellular free Ca2+
lular Ca2+or H+ concentration. Uncoupling of cells from and H+. The mechanism of closing of the gap junction
their neighbors can be produced by injecting Ca2+or H+ channel is not clearly understood, but the channel appears
to be open or shut depending on the relative positions of
the six subunits of the channel.
r Fluorescein
Distance
Tight Junctions
Figure 4-34 Gap junctions between coupled cells can be demonstrated Tight junctions seal cells together in an epithelial cell sheet
by following the flow of fluorescent dye injected into one of a group of so that even small molecules cannot get from one side of the
coupled epithelial cells. Subsequent diffusion of the dye into neighbor-
ing cells without loss into extracellular space indicates that there are di-
sheet to the other. The two apposing cell membranes make
rect pathways from the cytoplasm of one cell to the cytoplasm of the ad- intimate contact, fully occluding the extracellular space in
jacent cell. between. Tight junctions are found most commonly in ep-
Figure 4-35 Adjacent ep~thelialcells like
those that line the mammalian small intestine
are connected by intercellularjunctions. The
membranes and associated structures are
drawn disproportionately large in this recon-
Zonula occludens struction of the cell-celljunctions.
Zonula adherens
(intermediate junction)
Macula adherens
Intercellular space
ithelial tissues as a zonula occludens, a thin band of protein not preferentially transport solutes and water. In other
molecules that encircles a cell like a gasket. The zonula oc- cases they are involved in active transport, performing
cludens is in tight contact with the zonulae of the sur- regulatory functions. For example, osmoregulatory activ-
rounding cells, forming an impermeable seal that pre- ities of animals are carried out by actively transporting
vents passage of substances from one side of the epithelium epithelia in a variety of specialized tissues and organs (see
to the other via leakage down along the sides of the cells e a p t e r 15).
(Figure 4-35). Considered en masse, the zonulae are con- Epithelia have several features in common. First, they
ceptually like a continuous rubber sheet, penetrated only by occur at surfaces that separate the internal space of the or-
the ends of the epithelial cells.3ubstances can pass through ganism from the environment. Included are the surfaces
the ends of the cells (the transcellular pathway), but not lining deep invaginations such as in the lumen of the in-
around them (the paracellular pathway). In tissues such as testine, which nevertheless comprise external space. Sec-
the mammalian small intestine, the gallbladder, and the ond, the cells forming the outermost layer of the epithe-
proximal tubule of the nephron, these zonulae are not fully lium are generally sealed together by tight junctions, which
continuous and thus not really very "tight." These tissues to varying degrees in different epithelia obliterate paracel-
are so leaky, they do not produce a transepithelial potential lular pathways between the serosal (inner) and mucosal
difference, even though their cells contain ion pumps capa- (external) sides of the epithelium (Figure 4-36). In epithe-
ble of generating transepithelial ion fluxes. Unlike gap junc- lia such as that of capillary walls, leaky junctions permit
tions, tight junctions appear to have no special channels for water and solute molecules to cross the epithelial layer by
cell-cell communication. diffusion within the passages that exist between the cells.
Two other types of cell junctions are shown in Such diffusion through paracellular pathways is not cou-
Figure 4-35: the zonula adherens and the desmosome serve pled to any metabolically energized transport mechanism,
primarily to aid the structural bonding of neighboring cells. so such passages allow only passive movement of water
and ions. Substances that are actively transported across
an epithelium must follow transcellular pathways, in
EPITHELIAL TRANSPORT which the cell membrane participates. Such substances
Epithelial cell sheets line the cavities and free surfaces of must cross the cell membrane first on one side of the cell
animal bodies and form barriers to the movement of wa- and then on the other. As discussed in the next section, the
ter, solutes, and cells from one body compartment to an- functional properties of the surface membrane of an ep-
other. Each organ or compartment within an animal has ithelial cell are dissimilar in some respects on its serosal
such a lining of surface cells. Some of these sheets serve and mucosal surfaces. This asymmetry is important to
only as passive barriers between compartments and d o epithelial active transport.
120 PRINCIPLES OF PHYSIOLOGY
...............................................................................
Exterior,
lumen, or
mucosal side
I,/
Variable
current
Current meter
-
Transcellular
path
Acellular
basement
membrane
Paracellular
path
between two-half-chambers
Figure
- 4-36 Substances cross epithelial layers by two pathways:. para-
.
cellular and transcellular. Active transport takes place only across cell
Figure 4-37 Afrog skin separates the two halves of this Ussing chamber.
membranes, suggestingthat all actively transported molecules follow the
Each half is filled with a physiologicalsaline or other test solution.The cur-
transcellularpathway
rent source is adjusted until the potential difference across the skin is
zero. Under those conditions, the current flowing through the circuit (and
thus through the skin) is equivalent to the rate of charge transferred by
Active Salt Transport across an Epithelium the active movement of sodium ions across the skin.
Energy-requiringtransport of ions from one side of an ep-
ithelium to the other has been demonstrated in a number of
epithelial tissues, including amphibian skin and urinary In 1947 Ussing reported the first experiments in which
bladder, the gills of fishes and aquatic invertebrates, insect two isotopes of the same ion were used to measure bidirec-
and vertebrate intestine, and vertebrate kidney tubule and tional fluxes (i.e., the simultaneous movement of that ionic
gallbladder. Much of the initial work on epithelial active species in opposing directions across the epithelium). The
transport was done on the skin of the frog. In amphibians, Ringer's solution in the outside compartment was prepared
the skin acts as a major osmoregulatory organ. Salt is ac- using isotope 12Na+,and the Ringer's in the inside com-
tively transported from the mucosal side (i.e., the side fac- partment was prepared using 24Na+.The appearance of
ing the pond water) to the serosal side of the skin to com- each of the two isotopes on the opposite side of the skin
pensate for the salt that leaks out of the skin into the was tracked over time. The two isotopes were switched
freshwater surrounding the frog. Similar uptake occurs in around in other experiments of the same type to rule out
the gut. Water that enters the skin because of the osmotic any effects due to possible (but unlikely) differences in
gradient between the hypotonic pond water and the more transport rates inherent in the isotopes themselves. In all ex-
concentrated internal fluid is eliminated in the form of a co- periments it was found that Na+ shows a net movement
pious dilute urine that is hypotonic relative to the body flu- across the skin from the outside compartment to the inside
ids (see Chapter 14). one. That the Na+ flux is the result of active transport is
Frog skin was first used in the study of epithelial trans- seen in the fact that it
port in the 1930s acd 1940s by the German physiologist Occurs without any concentration gradient, and even
Ernst Huf and the Danish physiologist Hans Ussing. In against an electrochemical gradient.
their procedure, a piece of abdominal skin several square
centimeters in area is removed from an anesthetized and Is inhibited by general metabolic inhibitors, such as
decapitated frog and placed between two halves of an Uss- cyanide and iodoacetic acid, and by specific transport
ing chamber (Figure 4-37). The dissection is very simple, inhibitors, such as ouabain.
since the skin of the frog lies largely unattached over an Displays a strong temperature dependence.
extensive lymph space. Once the skin is gently clamped Exhibits saturation kinetics.
between the two half-chambers, a test solution-for ex- Shows chemical specificity. For example, Na+ is trans-
ample, frog Ringer's solution (a solution that mimics the ported while the very similarly structured lithium ion
ionic composition of frog plasma) -is introduced, with is not.
the frog skin acting as a partition between the two com-
partments. The compartment facing the mucosal side of How can an active movement of ions be produced
the skin can be designated as the outside compartment and across a layer of cells contained in an epithelium? Adjacent
the one facing the serosal side as the inside compartment. cells of transport epithelium are intimately tied together
Air is bubbled through the two solutions to keep them well with tight junctions. Assume for the sake of simplicity that
oxygenated. this closeness eliminates all extracellular passageways for
MEMBRANES, CHANNELS, AND TRANSPORT 121
.......................................
the diffusion of ions between the two sides of the epithe- Mucosal side Epithelial cell Serosal side
lium. This would force all substances that cross the epithe- (toward exterior) of froQ skin (toward interior)
lium to traverse the epithelial cell membrane twice, first
crossing the membrane on one side of the cell and then
leaving through the membrane on the other side. Active
transport by this route requires that the surface membrane
of each epithelial cell be differentiated, so that the portion
of the cell membrane facing the serosal side of the epithe-
lium differs in functional properties from the portion fat-
ing the mucosal side. Experiments on frog skin in Ussing
chambers have provided several lines of evidence to
support a hypothesis of a differentiated membrane. For
example,
Ouabain, which blocks the Na+/K+ pump, inhibits Figure 4-38 Transepithelial sodium transport depends on a combination
transepithelial sodium transport only when applied to of both diffusion and active transport. In this model of an isolated frog
the inner (serosal)side of the epithelium. It is ineffective skin that has been bathed in Ringer'ssolution, sodium diffuses passively
on the outer (mucosal) side. Conversely, the drug down its concentration gradient into the cell from the mucosal solution.
K+ diffuses out of the cell into the serosal space as it is displaced by Na+
amiloride, a powerful inhibitor of passive carrier-facil- influx. In the face of these leaks, a Na+/K+exchange pump in the serosal
. itated transport, blocks Na+ movements across the skin membrane of the cell maintainsthe high internal K+ and low internal Na+
only when applied to the outer side of the skin. concentrations. [From Koefoed-Johnsenand Ussing, 1958.1
:
C H A I ' T E K
5
T H E PHYSICAL BASIS
OF NEURONAL FUNCTION
During the embryonic development of each neuron, cle fibers, displays the structural and functional features
dendrites and the axon grow out from the soma. Through- that characterize many neurons (Figure 5-2).The surface
out the life of the cell, the maintenance of processes de- membrane of motor-neuron dendrites and the membrane
pends on the slow, but steady, flow down the processes of of the soma receive signals from, or are innervated by, the
proteins and other constituents that are synthesized in the terminals of other nerve cells. Typically, but not always, the
soma. If an axon in an adult organism is badly damaged or soma integrates (sums)messages from the many input neu-
severed, it typically degenerateswithin a few days or weeks. rons to determine whether the neuron will initiate its own
In mammals, regeneration or regrowth of axons is limited response, an action potential (AP), also called a nerve im-
to nerves in the periphery of the body, whereas in cold- pulse. The axon carries an AP from its point of origin, the
blooded vertebrates, some regeneration also may occur spike-initiating zone, to the axon terminals, which contact
within the central nervous system (i.e., the brain and spinal skeletal muscle cells in the case of motor neurons. Typically
cord). Damaged neurons in some invertebrates readily re- the spike-initiating zone is located near the axon hillock,
generate and reinnervate their original targets. the junction of the axon and cell body, although in
many neurons the spike-initiating zone lies elsewhere.
Transmission of Signals in a Single Neuron Many axons are surrounded by supporting cells, which
A typical vertebrate spinal motor neuron, which has its provide an insulating layer called the myelin sheath (see
soma in the spinal cord and carries signals to skeletal mus- Chapter 6).
.......................................
Dendrites Presynaptic terminals Voltage-gated ion channels allow ions to move across
the cell membrane in response to changes in the electrical
field across the membrane. Neurons possess various types
of ion channels, each permitting passage of a particular
ionic species. The different types of ion channels found in
neurons are not distributed uniformly over the surface of
I
INTEGRATION
the cell. Instead, they are localized in different regions that
have specialized signaling functions. For example, the ax-
onal membrane is specialized for the conduction of APs by
virtue of its fast-acting, voltage-gated ion channels that se-
lectively allow Naf and K' to cross the membrane. In ad-
dition, the membrane of axon terminals contains voltage-
1
'I SPIKE
INITIATION A
gated Ca2+channels and other specializations that allow
neurons to transmit signals to other cells when APs invade
the terminals.
'
.' Terminal
ganglion
segments
A
Nerve
cord
\
Metathora
gangl~or
rons In the central nervous system, and motor
(efferent) neurons controlling the muscles of
the legs. The wind-receptor neurons contact
the giant interneurons at synapses in the ter-
,/' Synapse minal ganglion ofthe nervous system, and the
giant interneuronscontact the leg motor neu-
rons at synapses in the thoracic ganglia. Stim-
ulation of the wind-receptor neurons causes
the cockroach to run away from the st~mulus.
vvlna recepror I
,
win'
,,.
neuron (afferent)
,i Giant
d
receptors Synapse
Sensory neurons, which transmit information collected cell bodies, called ganglia, distributed along the nerve cord;
from external stimuli (e.g., sound, light, pressure, and these control local regions of the animal (Figure 5-5).Ver-
chemical signals) or respond to stirnuli inside the body tebrates also have ganglia, consisting of peripheral neu-
(e.g., the blood oxygen level, the position of a joint, or ronal cell bodies, outside the CNS. In vertebrates the nerve
the orientation of the head) cord, called the spinal cord, is located along the dorsal mid-
Interneurons, which connect other neurons within the line, whereas in many invertebrates (e.g., insects, crus-
central nervous system taceans, and annelids), the major nerve cord lies along the
ventral midline. Many neurons that have cell bodies in the
Motor neurons, which carry signals to effector organs,
CNS send processes into the rest of the body (the periph-
causing contraction of muscles or secretion by gland cells
ery) to collect sensory information or to deliver motor in-
The essential function of sensory neurons is to transform formation to control the activity of muscles or glands.
the physical energy available in a stimulus into the electri- The other main cell type in the nervous system, glial
cal signals used by the nervous system. Networks of in- cells, fills all of the space between neurons, except for a very
terneurons, which are the most numerous type of neuron, thin extracellular space (about 20 nm wide) that separates
exchange information and perform most of the complex the glial and neuronal membranes. In general, more com-
computations that produce behavior. Motor neurons con- plex animals have a larger number of glial cells relative to
stitute the output portion of a neuronal circuit, carrying neurons than d o less con~plexanimals. The vertebrate
specific instructions to muscles or to other effector organs central nervous system, for instance, contains 10-50
that they innervate. (Neurons that innervate gland cells and times more glial cells than neurons, and these cells occupy
other effector targets are also called "motor" cells, even about half the volume of the nervous system. The
though they do not control bodily movement.) ratio of glial cells to neurons is considerably smaller
Neurons are grouped into clusters in almost all phyla. in most invertebrates.
Typically, the cell bodies of many-even most-neurons Glial cells provide intimate structural, and perhaps
are contained within the central nervous system (CNS),al- metabolic, support for neurons. Several different cell
though the cell bodies of some neurons are located in the types function as glial cells. In vertebrates, for example,
periphery. In most animals, the CNS consists of a brain, lo- oligodendrocytes in the CNS and Schwann cells in the pe-
cated in the head, and a nerve cord, which extends poste- riphery wrap each axon in an insulating myelin sheath,
riorly along the midline of the animal. Many invertebrates which contributes to assuring reliable and rapid transmis-
have a brain located in the head and collections of neuronal sion of APs (see Figure 5-2).
.......................................
Stimulus
Receptor
endings T-
....@
Sensory
GRADED
RESPONSE
..........................
ALL-OR-NONE
Afferent SIGNAL
/ \
Nerve cord (axons) Peripheral nerve trunk
Figure 5-5 The central nervous system, typically consisting of the brain
and nerve cord, is the site of most information processing and usually
contains most of an animal's neuronal cell bodies as well as many axons.
The brain, usually in the head of the animal, contains a large number of
neurons and their interconnections. In many anlmals, such as the medi-
cinal leech Hirudo medicinalisshown in this figure, the somata of other
neurons are grouped within the nerve cord, in structures called ganglia.
In a segmented animal like the leech, each segment commonly contains
a ganglion. Nerves containing axons connect structures within the CNS
and connect the CNS to peripheral structures.
\
Data analysis
I-
Original
signal
excitable tissues has a long history, which is reviewed in across the cell membrane (the membrane potential, V,) is
Spotlight 5-1. To understand both the basis for this ex- electronically amplified and displayed on a recording in-
citability and its consequences for how a neuron functions, strument. Until recently, recordings of membrane potentials
we need to be able to measure the details of the electrical were typically visualized using an oscilloscope, but re-
changes across the membrane as a function of time. searchers now rely extensively on digital computers to con-
trol equipment and display data during experiments, as
Measuring Membrane Potentials well as to analyze and store data later (Figure 5-6).
Electric currents are generated in living tissues whenever Much of what we know about how APs are generated
there is a net flux of charged particles across the membrane. rests on experiments carried out by A. L. Hodgkin and
Such currents can be detected directly by using two elec- A. F. Huxley in the 1940s and 1950s. They recorded mem-
trodes to measure the change in electrical potential that is brane potentials from squid axons, which are large enough
caused by current flow across the cell membrane. One sens- that a silver wire can be inserted and run longitudinally
ing electrode is placed in electrical contact with the fluid down the inside of this cylindrical process (Hodgkin and
inside the cell, and the other is placed in contact with the Huxley, 1952). The electrical activity of neurons that are
extracellular medium, so the two electrodes indicate the smaller than the squid giant axon has been studied using
voltage, or potential difference, between the cytosol and the glass capillary microelectrodes, which are described in
extracellular fluid. The potential difference measured Chapter 2. A cell is not damaged when it is impaled by a
microelectrode because the lipid bilayer of the membrane is the resting potential, V,,,,, of the cell membrane and is
seals itself around the pipette tip after penetration. Insertion most conveniently expressed in millivolts (mV, or thou-
of the electrode tip through the plasma membrane into the sandths of a volt). This resting potential is the membrane
cell brings the cell interior into electrical continuity with the potential measured when no active events or postsynaptic
voltage-recording amplifier. By convention, the membrane events are occurring. Virtually all cells that have been in-
potential is always taken as the intracellular potential rel- vestigated have a negative resting potential with a value be-
ative to the extracellular potential. In other words, the ex- tween -20 mV and - 100 mV.
tracellular potential is arbitrarily defined as zero. In prac- The potential sensed by the intracellular electrode does
tice the amplifier subtracts the extracellular potential from not change as the tip is advanced further into the cell, be-
the intracellular potential to give the potential difference. cause in the steady state the interior of the cell has the same
A simple arrangement for measuring membrane po- potential everywhere. Thus, the entire potential difference
tential is shown in Figure 5-7. In this experiment, the neu- between the cell's interior and exterior is localized across
ron is immersed in a physiological saline solution that is in the surface membrane and in the regions immediately ad-
contact with a reference electrode. Before the tip of the jacent to the inner and outer surfaces of the membrane.
recording microelectrode enters the cell, the microelectrode This potential difference constitutes an electrical gradient
and the reference electrode are both in the saline solution that is available as an energy source to move ions across
and are therefore at the same electrical potential; the po- the membrane. The electric field, E, equals the voltage in
tential difference recorded between the two electrodes is volts divided by the distance, d, in meters (E = Vld). Since
zero (see Figure 5-7A). As the tip of the microelectrode pen- d, the thickness of the membrane, is approximately 5 nm
etrates the cell membrane and enters the cytosol, a down- (5 x m), the actual electric field across cell mem-
ward shift of the voltage trace suddenly appears, indicating branes is very large.
that the electrode has entered the cell and is now measur-
ing the potential difference across the cell membrane (see Distinguishing Passive and Active Membrane
Figure 5-7B). By electrophysiological convention, inside- Electrical Properties
negative potentials are shown as downward displacements As noted earlier, the membranes of neurons and other ex-
of the oscilloscope trace. The steady inside-negative poten- citable cells display both passive and active electrical prop-
tial recorded by the electrode tip after it enters the cytosol erties. To understand the basis of electrical integration and
A Amplifier
Recording
Reference O mV
electrode
Saline
bath
-80 mV
Time -+
Recording
A Zinc B Zinc
Nerve st~mulated
when unl~ke
metals complete
clrcu~t
Current
Voltage
pulse Amplifier
Voltaae
display
-
d~splay
Current electrode
-- I-
-.Ic-c
Reference
electrode
Sal~nebath
,Ga
l ss capillary electrodes
/ filled with electrolyte
solution (e.g., 3 M K+ acetate)
inserted through surface
membrane.
Tip diameter of electrode is about 0
Figure 5-8 lmpal~nga cell w ~ t htwo m~croelectrodesallows the passlve Input reslstance, prevent~ngany apprec~ablecurrent from leavlng the cell
electrtcal propert~esof the membrane to be measured (A) Dtagram of an through the recording electrode (B) Magn~f~ed v~ewof the glass cap~llary
exper~mentalsetup to measure passlve membrane propert~esCurrent m~croelectrodesInserted through the membrane of the cell The elec-
flows in a c~rcu~t through the wlres, bath~ngsaltne, reslstor, current elec- trode at the left is used to pass current Into, or out of, the cell The cur-
trode, and cell membrane To malntatn constancy of the st~mulatingcur- rent changes the membrane potent~al,V,, as ~tcrosses the plasma
rent, the reslstor IS selected to have a far greater reslstance than the other membrane
elements of the st~mulatingclrcult. The recording ampl~f~er has a very h ~ g h
+40 r Figure 5-9 Some stlrnull evoke only passlve rnem-
brane responses, other stlmull may evoke actlve re-
sponses as well Shown here are typlcal records from
an experiment In which different amounts of current
are passed Into an excitable cell uslng the experl-
--
.-
m
mental setup deplcted ~nFlgure 5-8 The traces of
the stimulus currents (bottom) and the correspond-
o
- ~ n changes
g In the membrane potentlal (top) are In-
dlcated by small numerals lfthe ~ntroducedcurrent
causes the membrane potentlal to become more
negative wlth respect to the outslde of the cell, ~t1s
called hyperpolarlzlng (traces 1 and 2) If the Intro-
duced current causes the membrane potentlal to
become more posltive wlth respect to the outside of
the cell, ~tIS called depolarlzlng (traces 3 and 4) Hy-
perpolarlzlng currents and small depolarlzlng cur-
- rents produce passlve shlfts In V,,, (traces 1-3) Wlth
20
-s sufflclent depolarization, an addltlonal current enters
1 4
-
+
C
the cell, producing a sudden actlve electrical re-
sponse, the AP (trace 4a) Note that the slze of the
passlve responses IS more or less llnearly propor-
1 3 ??
L
tlonal to the stlrnulus current, whereas In the actlve
2
u2 0
-3
U)
response the change In membrane potentlal 1s much
U l z
.-
5
larger than would be expected for a h e a r response
At threshold, some st~mull will produce APs, whereas
other stlmull of the same magnitude will produce
-20 only a passlve response (trace 4b)
potential becomes less negative (e.g., it may shift from The active change in Vm that occurs in excitable cells
-60 mV to -50 mV). As the amount of applied current is depends on the opening or closing (gating) of numerous
increased, the degree of depolarization will also increase. ion-selective channels typically in response to depolariza-
Depolarizing a neuron causes membrane channels that are tion of the membrane. The opening (or closing) of a popu-
selectively permeable to sodium ions (Na+)to begin open- lation of ion channels controls the flow of an ionic current
ing. When electrically excitable cells become sufficiently de- that is driven from one side of the membrane to the other
polarized to open some critical number of Na+-selective by the electrochemical gradient for the ionic species per-
channels, an AP is triggered (Figure 5-9, trace 4a). The meating the channels. This gating of ion channels is the im-
value of the membrane potential that triggers production mediate cause for nearly all active electrical signals in living
of an AP is called the threshold potential. The opening of tissue. The simultaneous opening of many ion channels
voltage-gated Na+ channels in response to depolarization, generates the currents that are measured across cell mem-
and the resulting flow of Na+ ions into the cell, is one ex- branes. The voltage-gated ion channels that mediate active
- -
ample of membrane excitation. The opening of Na+-selec- electrical responses may be localized to particular areas
tive channels reduces the electrical resistance across the (e.g., the axonal membrane in neurons).
membrane (or increases the conductance), which can allow Most membrane channels allow one or a few species of
more current to flow. The mechanisms responsible for the ions to pass much more readily than all other ions; that is,
AP and other instances of membrane excitation are dis- they exhibit some degree of ion selectivity. The ion channels
cussed in more detail later in this chapter. that make cell membranes excitable are named for the ionic
species that normally moves through them. For example,
Role of Ion Channels Na+ is the ion that normally moves through the fast-acting,
Table 5-1 summarizes the properties of some ion channels voltage-gated sodium channels, which open in response to
that function in the passive and active electrical responses membrane depolarization, but certain other ions (e.g.,
of neurons. The passive change in V, that occurs in re- lithium ions) also can pass through these channels. In ad-
sponse to hyperpolarization or depolarization does not de- dition to voltage-gated channels, which respond to changes
pend on opening or closing of ion-selective channels. in V,,, , other ion channels are gated when messenger mol-
Rather, the ionic current that produces passive electrical re- ecules (e.g., neurotransmitters) bind to receptor proteins on
sponses flows primarily through K+-selectivechannels that the cell surface. These ligand-gated channels are discussed
are always open. These resting potassium channels are in Chapter 6. Still other ion channels, found in sensory re-
largely responsible for maintaining the resting potential, ceptor cells, are gated by specific stimulus energies such as
V,,, ,across the cell membrane. These channels typically are light (photoreceptors), chemicals (taste buds and olfactory
uniformly distributed over the entire membrane of ex- neurons), or mechanical strain (mechanoreceptors).These
citable cells. channels are discussed in Chapter 7.
TABLE 5-1
hGwn~leSof ion channels found in axons
Current through
Channel channel Characteristics Selected blockers Function
Restlng K+ channel (opsn Produces relat~velyhlgh Part~allyblocked by Largely responsible for V,,,
in resting axon) PKof rest~ngcell Tetraethylammon~um
(TEA)
Voltage-gated Na+ ha Rap~dlyact~vatedby Tetrodotox~n(TTX) Produces rlslng phase
channel depolar~zat~on,becomes of AP
inactivated even ~fV,,,
remalns depolar~zed
Voltage-gated Ca2+chan- I, Activated by depolariza- Verapamil, D600, Co2+, Produces slow depolariza-
nel tion but more slowly than Cd2+,Mn2+, Ni2+, La3+ tion; allows Ca2+t o enter
Na+ channel; is inacti- cell, where it can act as
vated as function of cyto- second messenger
plasmic [Ca2+]or V,
Voltage-gated K+ Activated by depolariza- Intra- and extracellular Carries current that rapidly
channel ("delayed tion but more slowly than TEA, amino pyridines repolarizes the membrane
rectifier") Na+ channel; is inacti- t o terminate an AP
vated slowly and not
completely if V,, remains
depolarized
Ca2+-activatedKt Act~vatedby depolar~za- Extracellular TEA Carr~escurrent that repo-
channel tlon and elevated cyto- lar~zesthe cell following
plasm~c[Ca2+],remalns APs based on e~therNaf or
open as long as cytoplas- Ca2+and that balances Ic,,
mlc [Ca2+]1s h~gherthan thus l ~ m ~ t depolar~zatron
~ng
normal by Ica
PASSIVE ELECTRICAL PROPERTIES portional to the current multiplied by the resistance of the
OF MEMBRANES membrane:
The capacitance and conductance of cell membranes,
AV, = AI xR (5-1)
which account for their passive electrical responses, corre-
spond to particular membrane structural elements. The
lipid bilayer, which is impermeable to ions, acts as an insu- where AV, is the voltage drop across the membrane, AI is
lator that separates charged ions, conferring on the mem- the current (in amperes) across the membrane, and R (in
brane an electrical capacitance. Ion channels, through ohms, a)is the electrical resistance of the membrane. The
which electric charges cross the membrane, give the mem- input resistance of a cell (i.e., the total resistance encoun-
brane its electrical conductance. These two membrane elec- tered by current flowing into or out of the cell) is a function
trical properties can be represented by an equivalent circuit of its membrane area, A, because the membrane of a larger
in which an electrical capacitor is connected in parallel with cell will typically contain more ion channels than does the
a resistor (Figure 5-10). The resistor represents the con- membrane of a smaller cell. Therefore, when membranes of
ductance of the ion channels, and the capacitor represents different cells are compared, it is necessary to take into ac-
the capacitance of the lipid bilayer. Such an equivalent cir- count the effect of membrane area. To do this, we define the
cuit is useful for making an explicit model of measured cur- specific resistance, R,, of the membrane as
rent flow through a membrane to determine likely values
for membrane conductance and capacitance. R,=RXA (5-2a)
Membrane Resistance and Conductance where A is the membrane area and Rmis the resistance of
The resistance of a membrane is a measure of its imperme- a Unitarea of membrane. Rearranging law gives
ability to ions, whereas the conductance is a measure of its
permeability to ions. For a given transmembrane voltage,
the lower the resistance of the membrane (i.e., the greater R = - AVm (5-2b)
AI
its conductance), the more ionic charges will cross the
membrane through ion channels per unit time. The rela-
tionship between current, resistance, and steady-state volt- then substituting yields
age across a membrane is described by Ohm's law, which
states that the voltage drop produced across a membrane R, = -x A (5-2~)
by a current passed across the membrane is directly pro- AI
138 PHYSIOLOGICAL PROCESSES
.......................................
where AVm/AI is in ohms and area is in square centimeters; Even though a membrane may be permeable to ion X,
thus Rmis in units of ohms cm2.Note that membrane area the conductance, gx,depends on the presence and concen-
and input resistance, R, are reciprocally related. The spe- tration of this species in the solution, because the ion
cific resistance, R,, of the membrane is a property of the species cannot carry current unless it is actually present. In
population of ion channels carrying ionic current across the addition, the permeability of the membrane to nonelec-
membrane. Specific resistances of various cell membranes trolytes does not contribute to conductance, because non-
range from hundreds to tens of thousands of ohms cm2. - electrolytes are uncharged and hence cannot carry current.
The reciprocal of resistance, R, is conductance, g (in Thus, the terms conductance and permeability are not
units of siemens, S): strictly synonymous.
Membrane Capacitance
Although ions do not cross the lipid bilayer except through
ion channels, they can interact across the membrane, which
Substituting this expression into Ohm's law gives is a thin insulating layer, to produce a capacitative current
(or charge displacement) even when no charges physically
cross the membrane in the process. When a voltage is ap-
plied across a membrane, positive ions will tend to move
towards the negative (cathodal)side from the positive (an-
The reciprocal of the specific resistance of a membrane odal) side in response to the force of the applied electric
is the specific conductance, gm (in units of siemens. ~ r n - ~ ) . field. Because the ions cannot cross the lipid bilayer, they
Conductance is closely related to the ionic permeability of pile up on the two surfaces of the membrane, cations on the
the membrane, but is not strictly synonymous with it. Con- anodal side and anions on the cathodal side, producing a
ductance for a given species of ion is defined by Ohm's law net excess of oppositely charged ions on opposite sides of
as the current carried by that species of ion divided by the the membrane. The membrane thus stores charges in the
electrical force acting on that species: same way that charges are stored by a capacitor in an elec-
trical circuit (Figure 5-11). Oppositely charged ions that
I,
gx = emf
where gx is the membrane conductance for ion species Equal Equal Equal
X, Ix is the current carried by that species, and emf is the +, - +. - +. -
electromotive force (in volts) acting on that species. Al- -A-
though emf varies with membrane potential, it is not iden-
tical with membrane potential, as will be discussed later.
Extracellular fluid
00 0
0 @ 0
core
Lipid
1 1 1 o n channel
(conductance)
Cytosol A
I
Membrane
Extracellular
fluid
(capacitance) Figure 5-1 1 The cell membrane acts as a capacitor. The cell membrane
can separate charges, with cations and anions forming a diffuse layer on
Cytosol opposite sides of the membrane and interacting electrostaticallyacross
the thin lipid barrier. This electrostatic interaction holds the charges in a
Figure 5-10 The passive electrical properties of a cell membrane can be narrow region immediately adjacent to the two surfaces of the cell mem-
represented by a simple electrical circuit. The membrane has a capaci- brane. Except for these few excess cations or anions on each side, the
tance because the lipid core is an insulator.The membrane's ionic con- bulk solutions on either side of the membrane conform to the principle
ductance depends on the presence of open ion channels. Arrows indi- of electroneutrality, as does the shaded region in the neighborhood near
cate flow of capacitative current, I,, and resistive current, I,. the membrane.
have accumulated on the two sides of the membrane can in-
teract electrostatically with one another because the mem- Current source
brane is so thin.
The ability of the bilayer to separate or store charges in
this way is its capacitance, which is measured in units of
coulombs per volt, or farads (F).The amount of charge that
can be separated by a layer of insulating material depends
on its thickness and on its dielectric constant, a property
that reflects the ability of an insulator to store charge. It is
possible to calculate an expected value for the capacitance
of nerve cell membranes if the membrane thickness and di-
electric constant of membrane lipids are known. Based
on a lipid-layer thickness of about 5 nm and a dielectric
constant of 3, which is about that of an 18-carbon fatty Membrane
acid, the membrane capacitance has been calculated to be
about 1 microfarad ( p F ) per square centimeter (1 pF =
1 0 F). Indeed, measured values of the capacitance of
biological membranes have proven generally to be close to
1p F . ~ r n - ~ .
The capacitance of a membrane affects how it responds
to a change in applied voltage or current. Positive ions pil-
ing up on one side of the membrane repel positive ions
from the other side, generating a transient flow of capaci-
tative current. Since the movement of charged particles near
the membrane takes time, capacitance effectively limits
how fast the voltage across a membrane can change under
specified conditions. This effect can be illustrated by an
Time -
Figure 5-12 The capacitance and resistance of the cell membrane are
responsibleforthe passive change in potential (electrotonic potential) in
equivalent circuit representing the neuronal membrane, as response to applied current. (A) Equivalent circuit for a cell membrane
shown in Figure 5-12A. In this example, a steady current, showing the current flow (colored arrows) when an abrupt, sustained
I,, of 1 ampere is applied suddenly to the membrane. Ac- pulse of constant current is passed across the membrane. (B) Time
courses for the currents and voltage produced by the susta~nedstimu-
cording to equation 5-la, such an applied current will pro-
lus shown in panel A. The top record shows how the total current is di-
duce a voltage drop across the membrane as shown in vided between the resistive current, Ir,and the capacitative current. I,.
Figure 5-12B (trace V,,,). This passive potential change, The middle record shows the membrane potential, Vm, (~.e.,the poten-
called an electrotonic potential, is produced by the current tial across both the membrane resistance and the membrane capaci-
that passes across the cell membrane. tance). The bottom record shows the total current across the membrane,
,I = Ic + IT.The time required for I, and Vm to reach 63% of their asymp-
As shown in Figure 5-1 2A, the current passing across
tot~c values is proport~onalto the product of the resistance and the ca-
the membrane must be distributed between the resistive pacitance of the membrane. This product is the time constant, r, of the
and the capacitative pathways, which are arranged in par- membrane.
allel across the membrane. When the steplike pulse of cur-
rent, Im,is forced across the membrane, the distribution of
current between the membrane capacitance and resistance
changes with time. At first, charges move relatively easily course (see Figure 5-12B). At the same time, the resistive
onto the capacitor component of the circuit, so most of the current, Illwhich passes through the membrane conduc-
current is carried by the capacitance as it charges, and little tance (i.e., through ion channels) increases exponentially,
current flows through the membrane resistance. The because the sum of Irand 1' necessarily equals the total cur-
buildup of charge on the capacitor is one form of electrical rent applied.
current, although no charges physically move across the ca- The relationship between potential and time during the
pacitor. Instead, the capacitative current, 5, is carried by charging of the capacitance is given by the equation
the displacement of charges onto, or off of, the opposite
sides of the lipid bilayer. As time passes and the capacitance Vt = Vz(l - CtlRC) (5-4)
charges, a voltage appears across the capacitance, which
slows the rate at which further charging occurs. This re- where Vx is the potential across the capacitor at time t = a
duction in the rate at which charges can move onto the ca- produced by a constant current applied to the network, t is
pacitor displaces an increasingly large fraction of current to the time in seconds after the onset of the current pulse, R
flow through the membrane resistance. Hence, the'capci- is the resistance of the circuit in ohms, Cis the capacitance
tative current, 1,., falls along an exponential time course, of the circuit in farads, and V, is the potential across the
while the membrane potential, V,, rises with the same time capacitor at any time, t.
When t is equal to the product RC, then V, =
V,(1 - lle) = 0.63 Vm.The value of t (in seconds) that
equals RC is termed the time constant (7) of the process.
Note that r is independent of both Vmand current strength.
It is the time required for the voltage across a charging ca-
pacitor to reach 63% of the asymptotic value, Vm(see Fig-
ure 5-12B, Vmtrace).
In summary, the passive electrical properties of the cell
membrane are its equivalent resistance and capacitance,
which are connected in parallel. Together they give the
membrane a time-dependent response to changes in volt-
age; the size and time course of the response depends on the
B
' Ion selective
membrane
ELECTROCHEMICAL POTENTIALS
All electrical phenomena in neurons and other cells de-
pend on the transmembrane potential difference, V,.
This voltage difference across the cell membrane is an
electrochemical potential that arises from two features
found in all eukaryotic cells. First, the concentrations of
several ions inside the cell are different from the concen-
trations of those same ions in the fluids outside the cell, and
these concentration gradients are maintained at the expense
of metabolic energy. Second, the ion channels that span the
membrane are selectively permeable to different ionic
species. These two properties result in the transmembrane
potential, Vm,which together with the electrical properties
of the membrane that we have just discussed produces the Figure 5-13 At electrochemical equilibrium, an electrical potential dif-
signals neurons use in communication. In this section, we ference exactly balances the chemical concentration gradient. (A) A
membrane that is permeable only to K+ separates compartments I and
will examine how the electrochemical membrane potential II, each of which contains 0.01 M KCI. (B) When the KC1 concentration in
arises and what determines its magnitude. compartment I is increasedto0.1 M, there is a small net movement of K+
To begin, consider the situation depicted in Figure 5-13. into compartment II. (C)The electromotive force (emf) corresponding to
A chamber containing 0.01 M KC1 is divided into two the resulting charge separation acts to impede further movement of KC.
compartments by a membrane permeable only to K+ ions. When the emf just balances the concentration gradient, the net move-
ment of K+ again becomes zero.
Under these conditions, the voltage recorded across the
membrane is zero. This hypothetical membrane is perme-
able to K+ but not to C1-, so K+ diffuses across the mem- indefinitely, provided there is no leakage of C1- across
brane without its counterion (the species of ion that coun- the membrane.
terbalances its positive charge, in this case C1-). However, How do we explain the maintenance of this potential
the net flux of K+ is zero, because the concentrations in the difference?After we increase the KC1 concentration in com-
two compartments are equal, so on the average, for each partment I, for every K+ that statistically is available for dif-
Kt ion that passes in one direction across the membrane, fusion through K+ channels from compartment I1 to com-
another will pass in the opposite direction. As a partment 1, 10 K+ ions in compartment I are available to
result, there will be no net flux of K+ ions, and the trans- pass across 5 2 membrane into compartment 11. The differ-
membrane potential will be zero (see Figure 5-13A). If ence in K+ concentrations-a chemical gradient, or chem-
we now increase the concentration of K+ in compartment ical potential difference-causes an initial net diffusion
I tenfold to 0.1 M, K+ will diffuse from compartment I to across the membrane from I to 11. However, each additional
compartment 11, producing an increase in positive charge K+ that diffuses from compartment I to compartment I1
in the latter (see Figure 5-13B). A positive potential will adds its positive charge to compartment 11, because C1-
thus quickly develop in compartment 11, and the voltmeter cannot accompany K+ across the membrane. As K+ ions
will indicate a potential difference between the two com- accumulate in compartment 11, the potential difference
partments (see Figure 5-13C). This potential difference across the membrane quickly rises, because the
will reach an equilibrium value that will be maintained membrane then separates an excess of positive charges on
one side from an excess of negative charges on the other (see the diffusible ion, and importantly, on the ratio of concen-
Figure 5-11). As K+ enters compartment I1 and produces trations on the two sides of the membrane:
this buildup of positive charge in that compartment, further
movement of K+ becomes less likely because extra positive
charges in compartment I1 repel additional positively
charged ions, while the electrostatic attraction of the excess
negative charges in compartment I holds K+ back. Here R is the gas constant, T is the absolute (Kelvin) tem-
In this situation, every K+ entering the membrane via perature; F is the Faraday constant (96,500 coulombs/gram-
a K+ channel has two forces acting on it: (1)a chemicalgva- equivalent charge);z is the valence of ion X; [XI, and [XI, are
dient favoring a net flux of K+ from I to 11, and (2)an elec- the concentrations (more accurately, the chemical activities)
trical potential difference favoring a net flux of K+ from I1 of ion X on sides I and I1 of the membrane; and Ex is the
to I. After some time, the opposing forces come into equi- equilibrium potential for ion X (potential of side I1 minus
librium and remain balanced, with the electromotive force side I). At a temperature of 18"C, for a monovalent ion, and
due to the electrical potential difference across the mem- converting from In to log, the Nernst equation reduces to
brane precisely offsetting the tendency for K+ to diffuse
down its concentration gradient (see Figure 5-13C). At this 0.058 [XI,
point, K+ is in electrochemical equilibrium. A potential dif- Ex = -log -
z [XI11
ference across the membrane that is established in this way
is called the equilibrium potential for the ion in question- where Exis expressed in volts. (At 38"C, which is approx-
in this case, the potassium equilibrium potential, E,. imately the body temperature of many mammals, the mul-
Once an ion is in electrochemical equilibrium, it shows tiplication factor is 0.061/z.) Note that Ex will be posi-
no further net flux across the membrane, even if the mem- tive if X is a cation and the ratio of [XI, to [XI,, is greater
brane is freely permeable to that ion. Conversely, if an ion than unity. The sign will become negative if the ratio is less
present in the system cannot diffuse across the membrane, than 1. Likewise, the sign will be reversed if X is an anion,
its presence does not influence the equilibrium state. Thus, rather than a cation, because z will be negative.
in our hypothetical system, C1- would contribute nothing By convention, the electrical potential inside a living
to the membrane potential, even though it is far out of elec- cell, V,, is expressed relative to the potential outside the
trochemical equilibrium (its concentration gradient would cell, Vo. That is, the membrane potential, V,, is given as
favor movement from compartment I into compartment V, - Vo, so the potential of the cell exterior is arbitrarily
11),because it is unable to cross the membrane. defined as zero. For this reason, when determining the equi-
It is important to note that the process of establishing librium potential across the cell membrane, we place the
the equilibrium state involves the diffusion of only a very extracellular concentration of the ion in the numerator and
small number of ions across the membrane, from one com- the intracellular concentration in the denominator of the
partment to the other, compared to the total number of ions concentration ratio. Applying the Nernst relation (equation
present in solution. Virtually no change takes place in the 5-6),we can calculate the potassium equilibrium potential,
concentvations of KC1 in the two compartments during this E,, in a hypothetical cell in which [KIo = 0.01 M and
process, because the number of KC ions that crossed into [K], = 0.1 M:
compartment I1 is insignificant compared to the number
originally present in the solution. This situation is discussed
further in Spotlight 5-2.
A QUANTITATIVE
There is one Faraday o f charge (= 96,500 coulombs) in 1 gram-
CONSIDERATION OF
equivalent weight (I mole) o f a monovalent ion. Thus t h e num-
CHARGE SEPARATION ber o f moles o f K+ required t o transfer 5.8 x c o f charge
across 1 cm2 o f membrane is calculated as follows:
ACROSS MEMBRANES
5.8 x lo-@C . C ~ - ~
It takes only a small number o f Ions d ~ f f u s ~ nacross
g 1 cm2 o f the =6 X 10-l3 m o l Kt p e r cm2
9 65 x 1O4 C.(mol K+)-l
membrane In F ~ g u r e5-13B t o allow the membrane p o t e n t ~ atlo
reach E,. In fact, the actual number o f excess ions that cross the
The number o f excess K ions that have accumulated in com-
membrane can b e calculated for a system that contains only one
partment II at equilibrium in Figure 5-13C is found b y multiplying
diffusible ion. The number o f excess K+ ions that accumulate in
the number of moles of K+ b y Avogadro's number (6 x lP3mol-
compartment II (and hence the number o f excess CI- ions left
ecules/mol):
behind in compartment I) depends o n t w o factors: E, and the ca-
pacitance, C, of the membrane. The charge, Q, that accumulates
across a capacitor equals the capacitance times the voltage, V: (6 x 1 0 - l ~m o l K+)(6 X l o z 3 ) = 3.6 X 10'' K+ per cm2
B
r
0-
-20
E
-40
C
.a,
0
,- 0 bsewed
values
C
s!
111
$ -80
I Nernst equation ( E K )
-100 - -0
-0
-120
0 0.2 0.5 1.0 2.5 5 10 20 50 100
External potassium concentration (mM)
Figure 5-14 The concentration ratio of K+ across the cell membrane af- ternal K+ concentration. The values of ENa,, were calculated from the
fects both the calculated membrane potential and V,,, measured exper- Goldman equation assuming PN, is 1% of P,, [K+], is 140 mm, and ignor-
imentally. (A) Semilog plot of the relationship between the equilibrium ing any contribut~onmade by CI-. The straight line represents the pre-
potential for K+, E,, and the ratio of K+ concentrations on the two sides dicted change in E, as [K+l0 changes calculated from the Nernst equa-
of a membrane, [K]o/[K],, calculated from the Nernst equation At 20"C, tion. Note that both the relationshippredicted by the Goldman equation
the slope of the line is 58 mVfor every tenfold increase In the concentra- and the experimental Vre, values deviate from this straight line at low
tion ratio. (B) Semilog plots comparing dependence of the calculated [K+l0.At higher [K+],, the calculated E, and EN,,, are nearly identical. [Part
EN,, and the measured V, in frog muscle (observed values) on the ex- B adapted from Hodgkin and Horowiu, 1960.1
The Goldman Equation: Calculating the muscles do lie close to this value, but it is possible to test
Equilibrium Potential for Multiple Ions further the accuracy of the hypothesis that V, depends on
If there is a concentration gradient across the membrane for the diffusion of specific ionic species.
a particular ion, the Nernst equation gives the equilibrium
potential for that ion. However, the Nernst equation can be THE RESTING POTENTIAL
used for only one ionic species at a time. In contrast, essen-
tially all cell membranes are permeable to several ionic At equilibrium, every cell that is in a nonexcited or "rest-
species, all of which are distributed asymmetrically across ing" state has a potential difference, Vrest,across its mem-
their plasma membranes. If more than one ionic species can brane. Typically, VI,, lies between -30 and -100 mV, de-
permeate the membrane, all of the permeating ionic species pending on the kind of cell and on the ionic environment.
.may contribute to setting the potential difference across the Two factors govern this potential: first, ion channels in the
membrane. In these more complicated situations, the membrane that are permeable to some-but not necessar-
Nernst equation does not apply, and a different equation ily all-of the ionic species present, and second, the
must be used to calculate a predicted equilibrium potential. unequal distribution of inorganic ions between the cell in-
Based on the notion that the transmembrane potential terior and cell exterior, maintained by active transport
is directly related to ion permeability, D. E. Goldman across the membrane and by the Donnan distribution (see
(1943) derived a quantitative representation of the mem- Chapter 4). The unequal distribution of ions provides a
brane potential when more than one ionic species can cross chemical driving force allowing an equilibrium potential to
the membrane. The Goldman equation can be considered be established.
as an approximate generalization of the Nernst equation Role of Ion Gradients and Channels
that has been extended to include the relative permeability
of each species of ion: As predicted by the Goldman equation, ions influence the
potential across a membrane roughly in proportion to the
permeability of the membrane to each of the ionic species
present. In the limiting case, if the membrane is imperme-
able to a particular ion species (as it is to the many large or-
ganic ions inside of cells), then the electrochemical gradient
of that species has no effect on the membrane potential, be-
in which P,, PNa,and PC,are the permeability constants for cause nonpermeating ions cannot carry charge from one
the major ion species in the intracellular and extracellular side of the membrane to the other. Conversely, if a mem-
compartments, and [K+],, and [K+],indicate the concentra- brane is permeable to only one species of ion, the distribu-
tions outside and inside the cell, respectively. tion of that ion will dictate the transmembrane potential,
In this equation, the probability that one ionic species which can be predicted by using the Nernst equation for
will cross the membrane is taken to be proportional to the that ionic species. If a membrane is permeable to more than
product of its concentration (more accurately, its thermo- one ion-and most biological membranes are-and the
dynamic activity) on that side and the permeability of the permepbilities are known, it is possible to predict the value
membrane to that ionic species. In frog muscle cells, the of V,,,, using the Goldman equation. In the previous sec-
permeability constant for sodium is about ?boo that of tion, we calculated a predicted EPP of -92 mV using mea-
potassium, and the membrane is nearly impermeable to sured values for the permeabilit~esand ion concentrations
chloride. In this situation, the Goldman equation can be in frog skeletal muscle fibers. In fact, measurements
simplified as follows: . of V,,,, in frog skeletal muscle fibers range from -90 to
- 100 mV, supporting the idea that the resting potential de-
Fershoot
+60 other types of exc~tablecells In vertebrate an~mals(e.g.,
heart muscle cells), each AP can last as long as half a second
5-
--
-
9
E
'--
(see Flgure 12-7).When the Interval between two APs 1s re-
duced, the second AP becomes progressively smaller, and lt
C
$2 falls completely lf a stimulus IS del~veredtoo soon after the
0a first AP. Durlng thls absolute refractory period the neuron
a
C
9
does not respond to a st~mulusof any slze.
a - - - - - - - - - - Threshold The flushlng of a tollet ~llustratessome of the features
~~~~l
E
5.
potentla1
of an AP. Once ~t 1s lnrt~ated,the flush (AP) continues to
complet~on,~ndependentof pressure (st~mulus) appl~edto
-80 - the trlggerlng lever. The flush IS thus an all-or-none phe-
3 -Threshold '~fter-hyperpolarization nomenon, like an AP. On the other hand, if after one flush,
2 n current the lever is pressed before the tank has filled completely, the
3 a - .1l l second flush that is produced will be smaller than normal.
If the lever is pressed too soon after one flush, a second
I I I I I I I I I I
1 2 3 4 5
flush may not occur at all.
In a neuron the situation is somewhat more compli-
Time (ms)
cated than in the flushing toilet analogy. N o stimulus, how-
Figure 5-16 A neuron produces an action potential when a stimulus ever large, is sufficient to evoke a second AP during the ab-
changes V , enough to exceed the threshold potential. The magnitudes solute refractory period, which exists during and for a short
of three stimuli are shown at the bottom of the figure, and the corre- time after an AP (Figure 5-17A). Following this period, the
sponding responses in a neuron are shown above. The small numbers
excitable cell membrane enters the relative refractory pe-
ind~catewhich stimulus produced wh~chresponse. Stimulus 3 depolar-
ized the membrane sufficiently to trlgger an AP. Smaller changes in V,
riod, during which the threshold potential is elevated above
evoked much smaller responses in the neuron (traces 1 and 2). The normal, but a strong stimulus may evoke an AP. An AP ini-
dashed curve illustrates the change in V , that would occur if the neuron tiated during this period may have a reduced amplitude (the
produced only a passlve response to stimulus 3, rather than an AP. overshoot is smaller). Excitability progressively increases
Sometimes a stimulus that IS just at the threshold value evokes an
(and the threshold potential decreases) during the relative
abort~ve,nonpropagated excitation, called a local response, rather than
an all-or-none AP
refractory period until it returns to the level characteristic
of the resting membrane before stimulation (Figure 5-17B).
Refractoriness, or diminished excitability, during and im-
produce passive depolarization until the current delivered mediately after the AP prevents fusion of impulses, but per-
is strong enough to depolarize the membrane to its thresh- mits the propagation of discrete impulses.
old potential, whereupon an AP is triggered. If the depo- If a neuron is stimulated by a series of subthreshold de-
larization is just too small to reach threshold, there may be polarizations, a time-dependent decrease in excitability oc-
an abortive, nonpropagated excitation called a local re- curs (i.e., the threshold potential increases). For example, if
sponse, which is simply the beginning of an AP that died the membrane is depolarized gradually with a current of
out before it was irreversibly under way. steadily increasing intensity, a greater depolarization is
The threshold current is the intensity of stimulating cur- required to elicit an AP than when the stimulus has an
rent that is just sufficient to bring the membrane to the abrupt onset (Figure 5-18).The slower the rate of increase
threshold potential and elicit an AP. Although most in the intensity of the stimulating current, the greater the in-
neurons have threshold potentials between -30 mV and , crease in threshold potential. This characteristic of ex-
-50 mV, no absolutely consistent value can be assigned ei- citable membranes, called accommodation, results from
ther to the threshold current intensity or to the threshold time-dependent changes in the sensitivity of membrane
potential, because the threshold depends on immediate past channels to depolarization.
electrical events that can modify the state of the membrane. When they are stimulated continuously by a current of
Once the threshold potential is reached, the AP be- constant intensity, some neurons accommodate rapidly and
comes regenerative; that is, the event becomes self-perpet- generate only one or two APs at the beginning of the stim-
uating, and Vmcontinues to change with no further stimu- ulus period (Figure 5-19A). These neurons are said to have
lus required. As the cell interior rapidly gains positive ions, a phasic response. Other neurons accommodate more
V, becomes less negative until the intracellular potential ac- slowly and therefore fire repetitively, although with grad-
tually exceeds zero (i.e., reverses polarity). The interior con- ually decreasing frequency, in response to a prolonged con-
tinues to become increasingly positive until it reaches a stant-current stimulus (Figure 5-19B). These neurons are
peak of +10 mV to $50 mV. The very brief period when said to have a tonic response. This difference among neu-
V,, is inside-positive is called the overshoot (see Figure rons plays a key role in how sensory neurons transmit in-
5-16). In mammalian neurons, APs typically last only a formation (see Chapter 7).The reduction in the frequency
millisecond or so, although in many invertebrate species, of APs that is typically seen in a neuron that responds ton-
APs can last as long as 10-or even 100-milliseconds. In ically during a sustained stimulus is termed adaptation.
Absolute refractory Relative refractory
period period
Time
A Phasic response
Time
Figure 5-17 During and after one AP, the neuron is refractory to pro-
ducing another AP. (A) Recorded changes in V, in response to three pairs Stimulus I
of stimuli that were delivered to a neuron. The timing of each stimulus is
represented by gray bars underthe V, traces; the thickness of these bars
indicate the strength of the stimulus required to produce an AP. In trace
1, the second stimulus produced no AP, indicating that it was delivered
during the absolute refractory period. In trace 2, the second AP was
B T o n ~ cresponse, l,
smaller and a stronger-than-normal stimulus was required to reach
threshold, indicating that the second stimulus was delivered during the vm
relative refractory period. When the two stimuli were separated by a suf-
ficiently long interval, both stimuli produced normally sized APs (trace 3).
(6) Time course of the change in membrane excitability during the re-
Stimulus s
fractory period. Durlng the absolute refractory period, the neuron cannot Time --)
be excited to produce another AP, regardless of the size of the stimulus.
During the relative refractory period, excitability is reduced (i.e., the Figure 5-19 Many, but not all, neurons adapt to sustained stimulation.
threshold is elevated), so stronger stimuli are required to reach threshold. (A) Some neurons exhibit strong adaptation to a prolonged stimulus,
Over time, membrane excitability returns to normal. producing only one or two impulses at the beginning of the stimulus, a
phasic response. (B)Other neurons adapt very little, except for a pro-
gresslve lengthen~ngof the ~ntersp~ke Interval, a tonlc response Note
of the features of the that we have that In t h ~ sf~gurethe overshoot of the APs was cut off by the record~ng
described so far can be explained at the level of the molec- device
ular structure of the membrane. Indeed, they all depend on
the selective regulation of ion channels in the nerve cell moves the membrane potential towards EN,. As the AP
membrane. reaches its peak, gNadeclines and gKincreases. Very quickly,
the outward K+ current exceeds the inward Na+ current,
Ionic Basis of the Action Potential causing the membrane potential to repolarize back toward
The electrical signature of an AP is the rapid depolarization E,. These dramatic changes in the ionic conductances re-
of the membrane. We now know that this change in Vmde- sult from molecular changes in the membrane. To summa-
pends on an inward Na+ current caused by a sudden large rize, at rest the membrane is most permeable to K+, but in
increase in the Na+ conductance (g,,) of the membrane the early phase of an AP, it becomes very much more per-
(Figure 5-20). During the rising phase of the AP, gN, ex- meable to Na+. When the permeability to Na' once again
ceeds g, (K+ conductance), and the net inward current becomes small, the membrane is at first very permeable to
- E Figure 5-20 An action potential is caused by
Na
transient changes in ionic conductances
across the membrane ln~t~ally gNaIncreases,
AP then w ~ t hsome delay g, ~ncreases,too The
AP In t h ~ f~gure,
s recorded from a squ~dglant
- 30 axon, conslsts of three phases a ris~ngphase
that depends upon the Increase ~n gNa,a
fall~ngphase that depends upon both a sud-
-
20 -2 den drop In g , and the rlse In g, and an af-
ter-hyperpolar~zat~on that occurs when g, re-
mains elevated for some tlrne The decrease
- 10 ~ngNa IS due to ~nact~vat~onof Nat channels,
whereas the decrease In gKIS caused by repo-
lar~zat~on When g,, IS h~gh,V, approaches
0 EN,,when gKIS h~gh,V, approaches EK
After-hyperpolanzat~on
- EK ....................................
w
K+ because voltage-gated KC channels are open. Later, the This equation says that there is an ionic current across the
permeability to K+ drops, because only the passive resting- membrane only if there is both a driving force and a con-
membrane K+ channels remain open. The permeability of ductance for ion X. If either gxor the emf for X is zero, there
the membrane to C I does not change during an AP. This will be no I,. For example, if many X-selective channels
brief overview of the mechanisms responsible for APs sum- were open, then gx would be high. However, if V, = Ex,
marizes decades of painstaking work by many scientists there would be no emf driving X across the membrane, and
who, through clever manipulation of the electrical and Ixwould equal zero. Likewise, if gxis zero (i.e., no ion chan-
chemical environment of axons, discovered the physical ba- nels for X are open), Ix will be zero regardless of the emf.
sis of signaling in the nervous system. What can be responsible for changes in ionic current
To understand how changing membrane characteristics across the membrane? To answer this, we can consider the
might account for the AP, it is useful to return to the model elements of equation 5-10. Ex,the equilibrium potential for
of a membrane as composed of a capacitance arranged in X, cannot change rapidly because it depends on the con-
parallel with a conductance (see Figure 5-12A), only we centration gradient of ion X, and the concentrations of X
will modify the model to include separate, parallel conduc- inside and outside the cell remain essentially unchanged
tances for each species of ion. These ionic conductances during an action potential. The variable in equation
represent open channels through membrane protein mole- 5-10 that is most likely to change is the ionic conductance,
cules, channels that carry the Na+ and KC currents across gx, which allows ion X to cross the membrane. Hence,
the membrane. Like any current carried by charged entities, changes in ionic conductance (i.e., changes in the number
ionic currents through membrane protein channels obey of ion channels that are open in the membrane) play a cru-
Ohm's law (equation 5-1). Thus, the current carried by ion cial role in controlling the electrical currents carried across
species X, I,, is given as biological membranes. Notice that changes in V, will also
affect the ionic current due to X, because the driving force
on X is equal to V, - Ex.
Our knowledge about how changes in the open state of
where gx is the membrane conductance for X, which is ion channels generate APs is due in large part to the efforts
proportional to the number of open X-selective channels, of several pioneering physiologists. In 1936, a distinguished
and emfx is the electromotive force acting on X. The elec- English zoologist, J. Z. Young, working at the marine sta-
tromotive force acting on X is the difference between the tion in Naples, Italy, first reported that longitudinal struc-
membrane potential, V,, and the equilibrium potential tures that had been described in squids and cuttlefish were
of X, Ex: not blood vessels, as had previously been thought, but were
instead very large axons (Figure 5-21A). These axons,
which control the speedy escape response of the animal, are
thought to have evolved their large size to facilitate rapid
If Ex differs from V,, there will be some driving force act- conduction of action potentials. They came to be known as
ing on X to force it across the membrane in one direction giant axons and proved to be a boon to biophysicists be-
or the other; the direction the ion will move can be inferred cause their large diameter-up to 1rnrn-allows electrode
from the sign of emfx. Substitutingthis expression for emfx wires to be inserted through them longitudinally for stimu-
into equation 5-8, we have a new form of Ohm's law: lation and recording (Figure 5-2 1B).
Working with giant axons, two groups of experi-
menters-K. S. Cole and H. J. Curtis in Woods Hole,
THE PHYSICAL BASIS OF NEURONAL FUNCTION 149
...............................................................................
A B Amplifier Figure 5-21 Giant axons of the squid, Loligo,
played an important role in early biophysical
studies of the AP. (A) Illustration of the squid
showing giant axons. Each stellate nerve con-
tains a giant axon that is several inches long
and has a d~ameterof up to 1 mm. Because of
their large size, giant axons conduct APs
rapidly and ensure the relatively synchronous
activation of all muscles in the mantle. When
-
recorded over time showing the AP as it
t
-
passed the location of the two electrodes il-
lustrated in part B. [Part A adapted from
Keynes, 1958.1
Stimulus
Stellate nerve
+
c s
--E
;+"[&--
C
(I)
(I)
C
2 -70
n
E
I
H
2 ms
- - - - - - - - - 1 mA out
compares Vmwith the command signal. If Vmis different from the preset 1 mA out
command potential, an electrical current is quickly passed across the 0
membrane to make V, once again equal to the command potential. If ----- --- 1 mA in
the permeabilityof the membraneto ions changes, more or less current
will be requiredto maintain V, at a constant level. (B) Results of a voltage-
clamp experiment illustrate the different effects of hyperpolarizingand -> bl +65 mV
(depolarization)
depolarizing voltage steps. When V ,was hyperpolarized(a),a small, con-
stant membrane current was induced (a').When V, was depolarized (b),
.5
(I)
F
-4t - - -- - - - 0
the induced ionic current initially was inward; later the ionic current was -+-
a r -65 mV
outward (b').(By convention, ~nwardfluxes of positive ions are plotted B I l l I I I (hyperpolarization)
downward from 0.) We now know thatthe early inward current is carried
by Na+ ions, and the later outward current is carried by K+ ions. [Part B 0 2 4 6 8 1 0
adapted from Hodgkin, Huxley, and Katz, 19521. Time (ms)
across the membrane into the cell. Later experiments neurons. However, these experiments could not reveal the
showed that the delayed outward current was carried by precise nature of these voltage-gated conductance channels,
K+ (see below). When the delayed current was subtracted the basis for their selectivity, or how they are activated. In
from the complex total current obtained in normal seawa- the time since Hodgkin and Huxley performed their ex-
ter, the difference between the two currents (shaded area, periments, two important advances have contributed sig-
Figure 5-23B) shows the time course of the inward current nificantly to our understanding of membrane channels.
carried by Nat, which is plotted in Figure 5-23C. First, techniques were developed for measuring ionic cur-
These experiments led to the hypothesis that a sudden rents across small regions of the cell membrane, even from
depolarization causes a significant number of Na+-selective one single ion channel. In addition, the techniques of pro-
channels to open briefly, producing an increase in the tein chemistry and molecular biology have made it possible
Na+ conductance through the membrane and allowing to identify the membrane proteins that constitute the chan-
Na+ to flow into the axon. In the normal extracellular nels. As a result, we now have a clear and consistent view
environment, the electrochemical gradient acting on Na+ of the molecular nature of ion channels, and we are devel-
(V, - E,) should drive Na+ into the cell. Thus, according oping a description of how modification in molecular struc-
to Ohm's law, when gNarises, so does INa : ture can allow the channels to change the conductance of a
membrane to particular ionic species.
To relate the molecular properties of channels to their
role in the generation of APs, we need to understand four
What does the time course of the Na+ current tell us key features of ion channels: (1)the distribution of ion
about the behavior of the membrane? Based on equation
5-11, the time course of INadepends both on changes in
/ channels in neuronal membrane; (2)the nature of current
flow through a single channel; (3)the mechanism by which
the conductance of the membrane to Na+, g,,, and depolarization of the membrane can open an electrically
on changes in the emf acting on Na+, which equals gated channel; and (4)the physical basis for how channels
V, - EN,. Clamping V,,, at a constant value holds the emf can select among ions. In this section, we will discuss each
on Na+ constant, because Na+ concentrations inside and of these features, particularly in relation to voltage-gated
outside the cell do not change. As a result, the time course Na+ channels.
of I, must directly reflect how gNachanges over time in re- Localization and characterization of voltage-dependent
sponse to depolarization. An important feature of I,,, il- channels has been facilitated by several naturally occurring
lustrated in Figure 5-23C, is that even when V,,, was held neurotoxins that bind to specific channels. One particularly
constant at a depolarized potential, INareached a maxi- potent and useful toxin is tetrodotoxin (TTX)from the vis-
mum within 1 ms and then rapidly returned to its low pre- cera of the Japanese puffer fish, Sphoeroides rubripes, and
stimulus value. Thus IN,, and hence g,, must consist of related species. TTX selectively blocks fast-acting, voltage-
two separate processes: activation, the time-dependent in- gated Na+ channels. When radioactively labeled TTX mol-
crease in g,, caused by a depolarization, and inactivation, ecules are added to the extracellular fluid, they bind to Na+
the time-dependent return of gNato its baseline level. channels. Examination of neurons labeled by this technique
Hodgkin and Huxley used the voltage-clamp technique has allowed the density of bound molecules, and hence of
to make time-dependent measurements of the distinct ionic Na+ channels, to be estimated. More recently, antibodies to
currents that contribute to the total current during an AP, the channel proteins have been developed, allowing the
concluding that each ionic current reflected a separate con- molecules to be labeled and viewed directly (see Chapter 2).
ductance through the membrane that was selective for the In nonmyelinated axons from a variety of different types of
particular ion carrying that current. Using these data, they neurons, the density of Na+ channels has been measured at
formulated equations to express each conductance as a about 500 Na+ channels per pm2; in these axons, the Na+
function of V,,, and time. These equations predicted the channels occupy about Yioo of the total surface area. Al-
electrical behavior of a nerve membrane under many dif- though this value might seem to be a surprisingly low den-
ferent conditions and demonstrated that properties of sity of channels, other calculations have indicated that each
many excitable membranes could be entirely accounted for channel can pass up to 10' Na+ ions per second, providing
by time-dependent changes in Na+ channels and K+ chan- enough I,, to account for the macroscopic currents that
nels in the membrane. have been measured in various neurons.
Observing the properties of current flow through a sin-
Voltage-gated sodium channels gle membrane channel proved difficult, because ordinary
Hodgkin and Huxley's detailed electrical measurements in voltage-clamp techniques cannot record currents through
squid giant axons revealed that gNaand gKchange during single channels. A conventional voltage-clamp device nec-
an AP. They hypothesized that these changes in conduc- essarily collects current from thousands of channels con-
tance permitted ions to move across the membrane and tained in a large area of membrane, and the precision of
that the resulting ionic currents caused the AP.Their elegant recording is limited by substantial background electrical
experiments clearly defined the aggregate, macroscopic noise arising from the passive flow of current through other
properties that must characterize membrane channels in membrane channels. However, in the late 1970s pioneering
T H E PHYSICAL BASIS O F N E U R O N A L F U N C T I O N 153
........................................
work by E. Neher and B. Sakmann led to the development on the charge, causing it to move in space, thus producing
of patch clamping, a technique for recording from individ- a conformational change in the molecule. Consider a typi-
ual ion channels using a modification of glass recording mi- cal resting neuron with a potential difference of about
cropipettes. In this method, a micropipette with a tip diam- -75 mV across the membrane. A depolarization of 50 mV
eter of 1-2 pm is placed in intimate contact with the (to -25 mV) generally activates a large fraction of the Na+
membrane and gentle suction is applied, producing a very channels present in such a membrane. These channels con-
tight, high-resistance seal between the pipette and' the cell sist of protein molecules inserted through the lipid bilayer
surface (Figure 5-24A; see also Figure 2-6). This powerful of the membrane, which is about 5 nm thick. It can be cal-
technical advance has profoundly influenced how we think culated that the portions of the channel proteins within
about membrane channels. The patch-clamp technique has the 5-nm layer of the membrane sense a voltage change
made it possible to record the activity of ion channels in the of 10-" per 10-"m, or 100,000 V-cm-', during the 50-
membranes of many types of neurons and from many mV depolarization. Charged groups on channel proteins
species. The results of these experiments has indicated a re- certainly might move, driven by this huge membrane elec-
markable amount of conservation throughout phylogeny. tric field and producing conformational changes in the
Neurons in animals as widely separated as jellyfish and mice proteins.
share ionic mechanisms that underlie their APs. When Hodgkin and Huxley proposed that membrane
Using the patch-clamp technique, researchers can depolarization would lead to a movement of the gating
record ionic currents through single membrane channels charge from inside to outside of the membrane, they also
while V,,, in that small region is clamped to a chosen value. suggested that this movement of charge should corre-
When a sufficiently large depolarizing step voltage is ap- spond to a small gating current (Ig)that would be associ-
plied, the recorded events occur as all-or-none currents hav- ated in time with the opening and closing of Na+ chan-
ing a square shape that indicates abrupt opening and clos- nels. For technical reasons, this hypothetical Ig was not
ing of the ion channel whose activity is being recorded detected until the early 1970s, when the development of
(Figure 5-24B). The currents are of similar amplitude very sensitive techniques allowed it to be measured. $ can
for individual channels that share a particular ion selectiv- be observed when the much larger ionic current through
ity and kinetics. The observed similarity in the records from the N a + channels, INa,is pharmacologically blocked by
channels with different ionic specificities suggests that tetrodotoxin or a similar agent. Gating currents also have
all voltage-gated ion channels function in the same gen- been detected for K + channels and for Ca2+ channels in
eral way. The time that individual channels remain open several tissues.
varies randomly over a wide range. The conductance of Figure 5-25 presents a model of the opening and clos-
a single sodium channel does not depend significantly on ing of a voltage-gated Na+ channel when a depolarizing
V,; its value ranges from 5 to 25 picosiemens, pS (10 pS = step voltage is applied to the membrane. Detailed analysis
1 0 x 10-l2 S, or lo1' Cl of resistance) for different of the Na+ channel gating current has rhvealed that (1)gat-
channels. ing of the channel takes place in several distinct steps, each
From Ohm's law, Faraday's constant, and Avogadro's of which is associated with charge movement, and (2)ac-
number, it can be calculated that one activated Na+ chan- tivation and inactivation of the channel are coupled
nel carries Naf ions at a rate of about 6000 ions per milli- processes. It appears that even though inactivation is not
second at an emf (V, - ENa) of - 100 mV (approximately associated with a gating current, the voltage dependence of
the driving force as the AP gets under way). The summed inactivation (see Figure 5-25, step e) occurs because the ac-
activity (i.e., openings and closings) of thousands of Na+ tivation and inactivation processes are coupled.
channels, each contributing a minute pulsatile unitary cur- Once a Na+ channel is open, only certain ions can pass
rent, gives rise to the macroscopic INa that produces the ris- through it. A channel's selectivity is indicated by its relative
ing phase of the AP (Figure 5-24C). The number of N a + permeability for various ion species. For instance, if the
channels open at any instant depends on time (because permeability of the Na+ channel for Na+ is set at 1.00,
of the time course of processes that lead to channel activa- than the permeability for Li+ is 0.93 and for K+ is only
tion and inactivation), as well as on V,. Thus macroscopic 0.09. The channel acts as if it contains a filter that selects
changes in the gNaof the membrane, which occur as func- partly on the basis of size, but estimates of channel size rel-
tions of V, and time, reflect the behavior of thousands of ative to the size of permeating ions suggest that relative size
Na+ channels, each one opening and closing during depo- cannot be the whole story (Figure 5-26A). Current hy-
larization in accord with certain probabilistic principles. potheses explaining how a channel selects among ions are
How could depolarization of the membrane influence based partly on ionic size and partly on other properties of
the opening of voltage-gated ion channels? Hodgkin and the permeating species. Negative charges located at the
Huxley originally suggested that changes in V, might reg- outer mouth of a cation-selective channel, such as Na+ and
ulate gNaand gKby causing a conformational change in a K+ channels, attract cations and repel anions. Cations
gating molecule. According to their proposal, a gating mol- A
larger than 4 in diameter are too large to pass through
ecule would bear a net charge at physiological p H values the pore of either Na+ or K+ channels. Cations smaller
and a change in membrane potential would produce an emf than 4 A pass through the pore, but only once they have
154 PHYSIOLOGICAL PROCESSES
...............................................................................
Current flows
when channels
are open
\ \ I 1 /, Suction produces \,
high-resistance seal \ p
-./iet I
/ / ,between glass and
\
\ 11
1 Cell m e h b r a n e
I
Depolar~zing
voltage step
Ensemble current
reconstructed by summing
many traces like those
in B
1
0
- 30
Time (rns)
60
Figure 5-24 The patch-clamp technique allows current through a single periments described in Spotlight 5-3. (B) A depolarization (black trace) of
membrane channel to be recorded. (A) A fire-polished patch pipette, a patch of rat muscle fiber membrane caused a single sodium channel to
with a tip diameter of about 2 p m and containing the same solution as open transiently several times, producing unitary sodium currents that
the one bathing the rest of the neuron, is sealed againstthe clean surface varied in duration and latency (three colored traces). (C) Summation of
membrane of a neuron until a very high-resistance contact is achieved, 144 such records from one patch produced an "ensemble current," the
which prevents the loss of current from the pipette to the outside saline. tlme course of which reflects the temporal distribution of individual chan-
Current flow through an open channel is detected by a sensitive elec- nel openings of that one channel following depolarization. This time
tronic circuit. The voltage across the patch of membrane that is sur- course resembles that of a macroscopic sodium current, which depends
rounded by the tip of the pipette is clamped using an electronic feedback on the activity of many channels in response to a single depolarizing step
circuit similar in principle t o that used in the original voltage-clamp ex- (see Figure 5-23C). [Parts Band C adapted from Patlack and Horn, 1982.1
tI T H E P H Y S I C A L BASIS O F N E U R O N A L F U N C T I O N
...............................................................................
Extracellular side
155
lnact~vat~ng gate
part~cle
Cytosol
-60 mV -
vm
-60 mV "OmVr - 4-L Y,IL
Figure 5-25 The conformation of the voltage-sensitive Na+ channel perimposed on an initial capacitative current, I=, through the membrane
changes during membrane excitation. A schematic model representing capacitance. kc) When the majority of the Na+ channels have opened, the
the opening and closing of the channel is shown, with cumulative current inward Na+ current, INa, is maximal. As depolarization continues (d),open
records corresponding t o each step below. In the resting state (a), the ac- channels begin t o close, because the inactivation palticle moves t o block
tivation gate is closed and the inactivating particle is located away from each open channel. After the membrane is repolarized (e), the gating
the pore. When Vm isvoltage-clamped t o a depolarized level (b), the gat- charges of the Na+ channel again reorient, giving rise t o another capac-
ing charge (curved arrow) moves in response t o the new electr~cfield itative gating current. The inactivation particle moves out of the channel,
across the membrane, producing a small gating current I,, and the ac- and the activation particle moves into the pore, returning the channel t o
tivation gate for the channel moves t o the open configuration. I, is su- its resting state (f).
lost the shell of water molecules (the water of hydration) time and no longer respond to depolarization. The spon-
that normally surround charged species in free aqueous so- taneous termination of the Na+ current by the intrinsic in-
lution (Figure 5-26B). How readily polar oxygen or activation of Na+ channels would be sufficient to finish
charged functional groups that line the pore of the channel an AP. However, K+ channels in the membrane accelerate
can substitute for the water of hydration determines how the recovery of the membrane potential following depo-
easily ions of the right size pass through the channel. Ac- larization.
cording to this hypothesis, some mechanism must exist in .
the Na+ channel for selectively replacing the water of hy- Voltage-gated potassium channels
dration surrounding Na+ ions, making the channel most The neuronal membrane also contains voltage-gated potas-
permeable to Na+. sium channels whose probability of opening is increased by
In summary, in an AP Na+ channels respond to an ini- depolarization. In comparison with the voltage-gated Na+
tial depolarization by opening, allowing Na+ to enter the channels, however, these K+ channels respond more slowly
cell, which further depolarizes the membrane. This depo- to voltage changes. The membrane gKdoes not begin to in-
larization causes more channels to open, allowing still crease until the AP is near its peak, and g, remains high
more Na+ to enter the cell and triggering an explosive, re- in the falling phase. The net outward current through the
generative evegt. This relationship between membrane K+ channels brings Vmback toward its resting value, and
potential and sodium conductance, termed the Hodgkin in neurons that have an after-hyperpolarization, the
cycle, represents a type of positive-feedback system membrane potential moves closer to EKthan it is at rest (see
(Figure 5-27). Such systems are rarely found in biological Figure 5-20).
tissue because they are inherently unstable. As noted pre- Many types of voltage-gated K+ channels do not in-
viously, once an AP is started, it needs no additional stim- trinsically inactivate, as do voltage-gated
- Na+ channels. In-
-
ulus to continue. The effects of this positive-feedback sys- stead, their conductance to ions depends only on Vm,so as
tem on the membrane are limited, however, in two ways. I, returns Vmback toward its resting value, g, decreases.
First, as the membrane potential approaches EN,, the As a neuron becomes depolarized, its K+ channels open,
driving force on Na+ is reduced. Second, open Na+ chan- thereby speeding repolarization. As the cell repolarizes, the
nels are inactivated, independently of V,, after a short K+ channels close again. As noted above, K+ channels are
156 PHYSIOLOGICAL PROCESSES
...............................................................................
Sodium channel Potassium channel
A
3.3 A
Selectivity
for ions Na+ > Li+ > Ca2+> K+ K+> Li+ > Na2
B + H,N-OH + H,N-NH,
LiC. H,O -
Na+ H,O Hydroxylamine Hydrazine
Figure 5-26 Ionic selectivity of channels is relatively specific and proba- channel may be rectangular, rather than round. The selectivity filter is
bly depends upon the relative size of some critical portion of the channel, thought to contact the dehydrated ions directly, so the channel must strip
along with the ease of dehydrating the permeating Ions. (A) Schematic the water of hydration from the ions. (B) Schematic diagrams of several
diagrams of the Na' and K+ channels based on the relative permeabili- partially hydrated inorganic and organic ions. All of these ions can pass
ties of various ions. The K+ channel is thought to have a round selectiv- through the Na+ channel, but only K+ can pass through the K- channel.
ity filter that is smaller than at least one dimension of the selectivityfilter The channel sizes and ions are drawn to the same scale. [Part B adapted
in the Na+ channel. The sizes and shapes of the organic ions that can from "Ion channels in the nerve cell membrane" by Rlchard D. Keynes.
pass through the selectivity filter of the Na+ channel indicate that the Copyright 0 1979 by Scientific American, Inc. All rights reserved.]
not needed by the membrane to generate APs, and indeed lowing neurons to generate APs at a higher frequency than
some myelinated mammalian neurons appear to lack them they otherwise could.
entirely. However, the acceleration of membrane repolar-
ization mediated by K+ channels does shorten the AP, al- Absolute and relative refractory periods
As we saw earlier, immediately following an AP the neu-
ronal membrane first is nonresponsive and then less re-
Applied outward current sponsive to stimuli than it is at rest (see Figure 5-17).These
absolute and relative refractory periods result from the
same mechanisms responsible for the after-hyperpolariza-
Membrane tion: inactivation of Naf channels and continued activation
F\
depolarization of K+ channels. The majority of Na+ channels that partic-
ipate in an AP remain inactivated, and thus cannot be
opened by depolarization, during the falling phase and for
a brief period thereafter. At a slightly later time-many, but
Na+ enters N a channels not all-of the Na+ channels are no longer inactivated, so
through channels open a depolarization can open a few channels. However, be-
cause this smaller number of channels produces much less
inward current, an above-normal depolarization is required
to generate enough inward current to initiate an AP during
this period. Moreover, outward current through the K+
channels that are still open from the previous AP also op-
Figure 5-27 The Hodgkin cycle is the positive-feedback loop between
poses the inward Na+ current. Together, these two ionic
membrane depolarization and sodium conductance responsiblefor the mechanisms produce the macroscopic effects measured as
rising phase of an A? The cycle is normally initiated by a depolarization refractory periods.
of the membrane that comes from outside the neuron and occurs inde-
pendently of the voltage-gated Na+ channels. The positive-feedback
Molecular structure of voltage-gated ion channels
loop usually is interrupted by the intrinsic inactivation of Na+ channels,
wh~chterminates the rising phase (black line 1). Alternatively, it is inter-
A combination of protein chemistry and molecular biology
rupted during a voltage-clamp experiment with the Na+ channels open has yielded detailed information about the channels that
(black line 2). conduct action potentials. Voltage-gated Na+ and K+ chan-
nels have now been identified, cloned, and sequenced in a among the various types of voltage-gated cation channels.
variety of species from Drosophila to mice and humans, A Na+ channel has been found to consist of one large a
and the homology among channels from different species is protein (about 260 kilodaltons, kD)that contains four ho-
striking. Furthermore, there is considerable homology mologous transmembrane domains (Figure 5-28A, left).
Extracellular sid
B Ca2+channel
c K+ channel / H5
- One subunit
Figure 5-28 The molecular structures of the voltage-gated Na+, K+, and channel complex. These small subunits, which differ between Na+ and
CaZ-channels are similar. (A,B) Each Na- channel and Ca2+channel con- Ca2+channels, contribute to the physiologicalproperties of the channels.
tains a large protein subun~t(a or a,) that can form functional channels (C) The cu subunit of a voltage-gated K+ channel is about one-quarter the
when expressed by itself in Xenopus oocytes (left). The asubunit in each size of the cu subunits in Na+ and Ca2+channels. It contains six a-helical
channel contains four homologous repeats (I-IV) of a-helical regions regions that are homologous to one repeat of the transmembrane heli-
(1 -6), each ofwhich is thoughtto span the lipid bilayer of the membrane. cal regions in the Na+ and Ca2' channel a subunits. Several variations of
The voltage sensor is thought to be located on the a helix numbered 4. the a-helical sequences are known for K+ channels. Afunctional K+ chan-
The four repeats of a-helical regions are thought to associate with one nel IS formed by association of four asubunits, which may have identical
anotherto form the ion-conducting pore of the channel (right). Each long or different sequences (r~ght).Smaller Psubunits that accompany the K+
cylinder In the channel diagrams correspondsto one repeat in the asub- channel asubunits in native neuronal channels help determine the phys-
unit. In addition, one or more smaller proteins (designated by various iological properties of these channels. [Adapted from Lodish et al., 1995,
Greek letters) normally associate with the asubunit to form the complete and from Hall, 1992.1
158 P H Y S I O L O G I C A L PROCESSES
.........................................
Although the a protein normally is associated with several ing potential, Vrc,,, of about 0.3 mV; following 10 action
smaller p proteins, it can, by itself, exhibit voltage-gated potentials in close succession, there is a cumulative depo-
channel function when it is expressed in Xenopus oocytes. larization of about 2 mV. Thus, the ability of small-diame-
The transmembrane domains of the cu protein are thought ter axons to continue generating APs depends on the rapid
t o be arranged around a central pore, forming the actual restoration of the intracellular and extracellular resting
path through which Na+ ions can flow (Figure 5-28A, concentrations of Na+ and K+ by active transport or other
right). Further analysis has identified particular amino acids mechanisms before cumulative ion fluxes produce signifi-
that are likely to confer the property of inactivation on cant changes in ion gradients. Note that the metabolic
these channels. pumping of ions across the cell membrane does not enter
This model of the Naf channel is strengthened by the directly into the production or recovery of an AP, but does
structure of the K+ channel protein. With a molecular mass serve to maintain the ionic concentration gradients required
of about 70 kD,the K+ channel protein is much smaller for the production of all membrane currents.
than the Na+ channel a protein, but it exhibits strong ho- Glial cells contribute to maintaining the VxSfof neurons
mology with the transmembrane domains of the Na+ chan- by taking up K+ from the extracellular fluids surrounding
nel cu protein. A K+channel is thought to be formed by as- axons. They then slowly release K+, allowing it to be re-
sociation of four of these smaller proteins in a manner very claimed by the neurons. Because the v,,, of all cells depends
similar to the way in which the four transmembrane do- heavily on the extracellular K+ concentration, recording the
mains of the cu protein associate to form a Na+ channel membrane potential of glial cells has provided a useful
(Figure 5-28C). Further molecular biological experiments measure of the changes in extracellular K+ concentration
employing site-directed mutagenesis are currently investi- that are produced when surrounding neurons conduct APs.
gating the details of ion selectivity, voltage gating, and other Changes in the Vnl of glial cells, which reflect the accumu-
properties of these voltage-gated channels involved in gen- lation of K-in extracellular spaces following neuronal ac-
erating action potentials. tivity, subside within several seconds, indicating that excess
K+ is quickly removed from the very restricted extracellu-
Changes in Ion Concentration during Excitation lar space. This removal is thought to depend, at least in
As mentioned earlier, only a few ions move across a cell part, upon uptake of K+across the surface membranes of
membrane to produce the voltage changes that occur dur- both neurons and glial cells. Thus glial cells probably help
ing a single AP, and they do not appreciably change the in- to prevent K t accumulation in the extracellular space,
tracellular ionic concentrations (see Spotlight 5-2). For ex- which would otherwise depolarize neurons.
ample, 10-12 mol of Na+ crossing 1 cm2 of membrane with
a capacitance of 1 p F cm2is sufficient to produce an AP of
100 mV amplitude. That is, only 160 Na+ ions per pm2 of
OTHER ELECTRICALLY EXCITED
membrane are needed. Since a single Na+ channel can pass
CHANNELS
l o 7 Na' ions per second, the lNarequired for an Transmembrane ion channels occur in virtually all cell
AP could be supplied by a remarkably small number of types. Although voltage-gated Na+ and K+ channels col-
channel openings. This calculated number of Na+ ions that laborate in the production of typical APs, Ca2+channels
cross the membrane during one AP is probably somewhat may be of more widespread importance in cell function (see
of an underestimate, because some K+ ions move out of Table 5-1).Ca2+-selectivechannels carry at least part of the
the cell, partially canceling the influx of Na+. The actual regenerative depolarizing current in crustacean muscle
number of ions is likely to be closer to 500 Na' ions/ fibers; in vertebrate smooth muscle cells and cardiac mus-
pm2/impulse. If this value is correct, during a single AP in a cle cells; in the cell bodies, the dendrites, and the terminals
squid giant axon with a diameter of 1mm, the intracellular of many nerve cells; in embryonic neurons; and in ciliates
Na+ concentration would change by only 1 part in more like Paramecium, to list just a few examples. In many of
than 100,000. For this reason, a squid axon can these membranes, Ca2+carries inward current along with
generate thousands of impulses after its Na+ pumps have Na+, but in a few cell types it carries all of the inward cur-
been incapacitated by a metabolic poison. Eventually, of rent. The ICatypically is not strong enough to produce an
course, in a poisoned axon the concentrations, and hence all-or-none AP without help from an INa,and in most mem-
the equilibrium potentials, of Na+ and K+ will change, and branes that contain voltage-gated Ca2+channels, the rising
the axon will no longer be able to sustain normal action phase of an all-or-none AP is generated largely by a strong
potentials. I,', that rapidly depolarizes the membrane. The Ca2+chan-
In smaller axons, which have a greater surface-to- nels, which open more slowly and conduct less current, are
volume ratio than the squid giant axon, a more significant activated by this depolarization. The Ca2+ions that enter
change in ionic concentrations occurs even following a sin- the cell through the Ca2+ channels often have two func-
gle AP. For example, mammalian C fibers have a diameter tions: propagating an electrical signal and acting as an in-
of only about 1pm; in these axons, ionic fluxes from a sin- tracellular messenger that triggers subsequent intracellular
gle impulse change the intracellular Na+ and K + concen- events. For example, Ca2+is responsible for the release of
trations by about 1percent. The result is a drop in the rest- neurotransmitter substances from the presynaptic terminals
and can also contribute to causing contraction of muscles
(see Chapters 6 and 10).
The molecular structure of voltage-gated Ca2+channels
is strikingly similar to voltage-gated N a t channels. Like
Na+ channels, Ca2+channels consist of a large protein ( a l )
that includes four homologous transmembrane domains,
which are thought to associate and form the ion-conduct-
ing pore similar to the Na- channel cu protein. The Ca2+
channel a , protein also typically associates with several
smaller proteins (Figure 5-28B). Embryonic neurons often
express both voltage-gated Na+ and voltage-gated Ca2+ SUMMARY
channels. Usually the Ca2+channels appear .
-
first, and the Specific electrical properties of the cell membrane affect the
Na+ channels become functional at later stages. The re- ability of neurons to carry information; these electrical
markably homologous molecular structure of Na+ and properties depend on the molecular makeup of the mem-
Ca2+channels, the frequency with which the two kinds of brane. The lipid bilayer membrane acts as an electrical ca-
channels appear in the same cells, and the greater preva- pacitance: although it does not readily allow charge carri-
lence of Ca2+ channels in simpler organisms such as the ers (i.e., ions) to pass, it is very thin (about 5 nm) and thus
protozoa suggest that Nat channels may be a more recent can accumulate and store charge by means of electrostatic
evolutionary specialization for impulse conduction. In ad- interaction between cations and anions on opposite sides of
dition, Ca2+acts as an intracellular messenger in most types the membrane. Channels composed of protein molecules
of cells, providing further evidence that Ca2' channels may embedded in the lipid bilayer provide selective electrical
have had a more primitive evolutionary origin than the conductances. These channels permit the physical passage
channels that pass monovalent cations. of certain inorganic ions across the membrane; the flow of
Unlike voltage-gated Na+ channels, many types of ions through such ion-selective channels constitutes an elec-
Ca2+channels fail to inactivate fully under maintained de- tric current. These two properties, capacitance and con-
polarization. Instead, in at least one type of Ca" channel, ductance, determine the time course of voltage changes that
the probability of inactivation increases as the intracellular are produced by the flow of current across electrically ac-
concentration of free Ca2+rises. These Ca2+channels be- tive cell membranes.
come inactivated during maintained depolarization because An asymmetrical distribution of ions in solution on the
ICathrough the channels increases the intracellular Ca2+ two sides of a membrane can produce an electrical poten-
concentration near the membrane. tial across the membrane, depending on how permeable the
The intracellular concentration of free Ca2+also regu- membrane is to the ions present. The size of the electrical
lates the function of a class of voltage-gated K + channels potential can be predicted from the Nernst equation or the
that differs from the delayed voltage-gated K+ channels Goldman equation. Because resting cell membranes are
that we discussed previously. These K+ channels, which are most permeable to K+and C1-, the resting potential usually
found in many different tissues, are activated by membrane lies close to the equilibrium potentials of these two ions,
depolarization, but only if the concentration of intracellu- typically between -40 and -100 mV (inside negative with
lar free Ca2+is higher than normal (see Table 5-1). In these respect to outside).
cells, Ca2+enters through Ca2+channels and accumulates Active transport of Na+ and Ca2+causes these ions to
near the inner surface of the membrane; it then causes these be less concentrated within the cytoplasm than outside the
Ca2+-dependentvoltage-gated K' channels to open if is cell. For each of these ions, there is a large driving force into
depolarized. Where these channels are present, the entry of the cell and a low permeability, so to maintain the low in-
Ca2+fosters repolarization as the result of the enhanced I,, tracellular concentration, they must be continually pumped
which carries positive charge (K+)out of the cell. This I, out. Stimuli that increase the normally low permeability to
also contributes to the production of an after-hyperpolar- either Na+ or Ca2+ lead to an influx of one or the other,
ization and to the refractoriness of neurons, thus setting which makes the interior of the cell less negative. For ex-
limits on the maximum frequency of APs that can be pro- ample, the transient opening of Na+ channels is responsible
duced by the neurons. for the rising phase (depolarization)of the action potential
Each of these four kinds of voltage-gated ion channels (nerve impulse). Since this increase in Na+ permeability is
is highly selective for a particular ion. The Na+ and Ca2+ evoked by membrane depolarization, the upstroke of the
channels normally carry current into the cell, because there nerve impulse is regenerative and causes the membrane po-
is a strong emf driving both ions into the cell. The delayed tential to approach briefly the sodium equilibrium potential
voltage-dependent K+ channel and Ca2+-dependent K+ of +50 to +60 mV at the peak of the nerve impulse. A de-
channel generally carry current out of the cell, because the layed rise in K+permeability is brought on by the voltage
emf on K+ drives it out of the cell. The distribution of these change across the membrane, and together with rapid in-
and other types of voltage-gated channels determine the activation of the Na+ channels, it brings the membrane
electrical behavior of excitable tissues. back to the resting potential, terminating the AP.
Thus the electrical behavior of excitable membranes de- What are two observations suggesting that Na+ car-
pends on the passive properties of membrane capacitance ries the inward current responsible for the upstroke
and resting conductance, on metabolically sustained ion of the action potential?
gradients across the membrane, and on the presence of ion- Does the sodium pump play a direct role in any part
selective membrane channel proteins, some of which are ac- of the AP? Explain. How is the sodium pump indi-
tivated by depolarization of the membrane. It is now pos- rectly important in the production of an AP?
sible to explain the details of changes in the membrane What limits the flux of NaT across the membrane
potential that occur during an AP in terms of the molecular during an AP? What limits the flux of K+ across the
properties of these ion channels. membrane during an AP?
Calculate the approximate number of sodium ions
entering through each square centimeter of axon sur-
REVIEW QUESTIONS face during an AP having an amplitude of 100 mV.
What are the major anatomic regions of a neuron, (Recall that 96,500 C is equivalent to 1mol-equiv of
and what is the principle physiological function of charge; that membranes have a typical capacitance
each? of F - c ~ - and
~ ; that Avogadro's number is
Does a signal that reaches the brain accurately en- 6.022 X atoms .molkl.)
code all features of the original stimulus? Why or The following properties of action potentials were
why not? discovered decades before physiologists knew about
The cell membrane separates electrical charge and ion channels and their role in action potentials.
therefore has a potential difference across it. Does Explain each of the properties in terms of the be-
this violate the general physical requirement for elec- havior of ion channels: (a) threshold potential;
troneutrality? Why or why not? (b) all-or-none overshoot; (c) refractoriness; and (d)
What is the structural basis for membrane capaci- accommodation.
tance? For membrane conductance? Why is it that an axon of large diameter undergoes
How is the time course of potential changes across essentially no change in ionic concentration during
the cell membrane related to the resistance and ca- several APs, whereas the very thinnest axons may
pacitance of the membrane? undergo significant changes in concentration dur-
Cells are typically inside-negative. Explain this ob- ing several impulses? How would these changes
servation in terms of diffusion potentials. affect the function of a small axon, and what mecha-
You have on your lab bench an artificial system of nisms might help to prevent the changes in ionic
aqueous solutions separated by a semipermeable concentration?
membrane. The solutions separated by the mem- The rising phase of an AP is an example of positive
brane contain the same ions, but in different concen- feedback in a biological system. How does positive
trations, and the membrane is permeable to only one feedback occur in this situation? Positive feedback is
of the ions in solution. Will there be a stable electri- inherently unstable, so how can you account for the
cal potential difference across the membrane? Why limited amplitude of the upstroke?
or why not? How do the properties of ionic currents through a
Living cells are typically quite permeable to K+ and single channel compare with the properties of macro-
at least slightly permeable to other cations as well. scopic ionic currents measured across the membranes
What maintains the high K+ concentration inside the of real cells?
cell by preventing the K+ inside from gradually being Name four kinds of voltage-gated ion channels that
displaced by other cations? contribute to neuronal function. How do channels
What are the equilibrium potentials for each of select for particular ionic species?
the following ions of the given concentrations?
(a) [K+], = 3 mm, [I<+],= 150 mm; (b) [Na+], =
100 mm, [Na+], = 10 mm; and (c) [Ca2+], =
10 mm, [Ca2+],= mm.
SUGGESTED READINGS
For a typical cell that is 100 times more permeable Aidley, D. J. 1989. The Physzology of Excztable Cells. 3d
to K+ than to any other ion, use the Goldman equa- ed. New York: Cambridge University Press. (A thor-
tion to determine the potential change that would ough examination of the physiology of nerves, muscles,
be produced by a doubling of the extracellular K+ and unusual features such as the electric organs of some
concentration. (Use concentrations from 9.) fishes.)
In 1939 Cole and Curtis reported that membrane Catterall, W. A. 1993 Structure and function of voltage-
conductance increases, but capacitance remains es- gated ion channels. Trends Neuroscz. 16500-506. (A
sentially unchanged, during an AP. Relate these find- readable review of recent work on the molecular fea-
ings to membrane structure and to changes in struc- tures of ion channels written by a major contributer to
ture thought to occur during excitation. the field.)
Hall, Z. 1992 An Introduction to Molecular Neurobiology. Hodgkin, A. L. 1976. Chance and design in electrophysi-
Sunderland, Mass.: Sinauer. (A description of neuro- ology: An informal account of certain experiments on
physiology that emphasizes the roles and properties of nerve carried out between 1934 and 1952. J. Physiol.
proteins and the regulation of their expression.) (London) 263:l-21. (An insider's view of biophysical
Hille, B. 1992. Ionic Channels of Excitable Membranes. 2d history.)
ed. Sunderland, Mass.: Sinauer. (An excellent com- Kandel, E. R., J. H. Schwartz, and T. M. Jessell. 1991. Prin-
pendium of information about the biophysics of ion ciples of Neural Science. 3d ed. New York: Elsevier. (An
channels.) enormous and authoritative compendium of informa-
Hodgkin, A. L. 1964. The Conduction of the Nervous Im- tion about the function of the nervous system, from the
pulse. Springfield, Ill.: Thomas. (A classic description of biophysics of membrane channels to the physiological
the electrophysiology of signal conduction in neurons.) basis of memory and learning.)
T he survival of all animals depends on their ability to re-
spond to challenges from other animals and from the
environment. Often, the response must be both rapid and
so, if the signal carried by any one neuron is to generate ef-
fective and adaptive behavior, it must be transmitted to
other neurons. Information is passed along to other neu-
well coordinated in order to be effective, and animals can rons at structures called synapses, and synaptic transmis-
produce such responses only when information is gathered, sion (i.e., the mechanisms that allow information to be
organized, and transmitted quickly throughout the body. passed from one neuron to the next at synapses) constitutes
Nervous systems evolved to allow such rapid and adaptive the second major topic of this chapter.
responses to occur. They are found in all animals, from the
simplest coelenterates to the most complex mammals.
TRANSMISSION OF SIGNALS IN THE
The complexity of nervous systems is amply illustrated
NERVOUS SYSTEM: AN OVERVIEW
by the human nervous system, which contains more than
1O1%eurons plus an even larger number of supportive cells Signals move from point to point along the plasma mem-
(glial cells, or neuroglia). The functional units that allow brane of a single neuron in either of two ways: graded, elec-
animals to respond effectively to their environments are sets trotonically conducted potentials and actiorz potentials
of neurons that are connected with one another in a way (all-or-none impulses). These two basic methods of trans-
that allows information to be passed among the cells. These mission alternate as information passes along one neuron
arrays are called neuronal circuits, their interconnections and is then transferred to another neuron, as illustrated
being in many ways analagous to electrical circuits. All the schematically in Figure 6-1, which shows the path traveled
complex capacities of the nervous system-movement, by information received at the interface between an animal
perception, learning, memory, and consciousness-arise and its surroundings (e.g., at the skin). Energy from a phys-
from the physical and chemical processes that were con- ical stimulus is received and changes the membrane poten-
sidered in Chapter 5 and are examined further in this chap- tial, V,, at a specialized type of membrane in a sensory
ter. Understanding how neuronal activity results in behav- neuron (see Chapter 7 for further treatment of this process),
ior is undoubtedly one of the greatest of all challenges to producing a receptor potential that is graded (i.e., varies in
biologists, and whether we will ever completely understand a continuous fashion) in proportion to the strength of the
the physical and chemical basis of consciousness and cre- stimulus. A low-intensity stimulus produces a small change
ative thought remains an open question. in V,,,; a more intense stimulus produces a larger change in
In spite of the enormous complexity of most nervous V,. This change in V, at the receptor membrane generally
systems, a great deal has already been learned about the continues, often with some attenuation, for the duration of
physiology and biophysics of single neurons, as described the stimulus. Because the time course and amplitude of a re-
in Chapter 5. We know that all neurons carry information ceptor potential are closely related to the time course and
by means of electrical signals that are based on the move- intensity of the stimulus, the receptor potential is an elec-
ment of particular ions across the cell membrane. Ionic cur- trical neuronal analog of the stimulus. For example, a pres-
rents (see Chapter 5) encode signals that travel along axons sure stimulus that persists for a long time typically produces
by mechanisms that constitute the first topic of this chap- a long, slightly attenuated depolarization in the receptor
ter. Although we consider events in single neurons when endings. This signal spreads along the cell membrane away
we examine the transmission of information by action po- from the location of the stimulus very much as an electrical
tentials (APs),behavior is never produced by the activity of signal spreads along a wire. The membrane that is special-
a single neuron. Even in very simple animals, behavior de- ized to receive sensory stimuli lacks the voltage-gated ion
pends on the activity of many neurons working together; chalinels that produce all-or-none APs, so signals cannot be
164 PHYSIOLOGICAL PROCESSES
U Stimulus
(mechanical,chemical,
from the regenerative transmission of APs. Passively trans-
mitted signals from receptor endings decay over relatively
short distances, so passive transmission is not effective for
light, etc.)
carrying signals to distant parts of the body. For long-
Receptor distance transmission, the sensory signals must be trans-
endings
formed into APs, which can conduct signals without
decrement for long distances. The membrane at the spike-
Graded receptor initiating zone of a sensory neuron contains the voltage-
Spike-initiating
potential
zone
(electrical)
gated ion channels that permit APs to start, so if the pas-
sively propagated depolarization of a receptor potential is
large enough at the spike-initiating zone, the signal will be
transformed from passive into active propagation and will
I I
All-or-none be carried without decrement along the sensory neuron's
impulses axon, which may span many meters.
(electrical)
The next transformation of the signal takes place at the
axon terminals of the sensory neuron, where it must be
passed across synapses to other neurons. This transfer of in-
formation between neurons is typically (but not always) ac-
complished through chemical signals carried by molecules
called neurotransmitters. If the transfer of information
across a synapse is to be effective, the neurotransmitter must
cause the membrane potential, V,, of the postsynaptic neu-
ron to change. The amount of transmitter released, and thus
the amplitude of the response in the next neuron along the
pathway, depends on the number and frequency of APs ar-
riving in the terminals of the sensory neuron (called the
presynaptic neuron). Within limits, higher frequencies and
more APs in the presynaptic neuron cause more neuro-
transmitter molecules to be released, producing a greater
change in V, of the receiver neuron (called the postsynaptic
neuron). The change in V, of the postsynaptic neuron,
called the postsynapticpotential (psp),is a graded signal, re-
flecting at least some properties of the original stimulus, al-
though it may be quite distorted. If it is sufficiently large, this
graded postsynaptic potential can bring the spike-initiating
Figure 6-1 Graded and all-or-none electrical signals alternate with one
zone of the postsynaptic neuron to threshold, triggering one
another as a signal is carried along a neuronal circuit. A stimulus applied
to the receptor endings of a sensory neuron produces a graded poten- or more all-or-none APs in the postsynaptic neuron.
tial that reflects the amplitude and duration of the stimulus, although it is Thus, as a signal is received from the environment and
neither l~nearlyproportional to the stimulus nor identical in form. This po- transmitted by neurons, it is coded alternately in graded po-
tential spreads passively through the first part of the sensory neuron and, tentials and in all-or-none APs. Graded potentials are pro-
if the change in V, is sufficiently large at the spike-initiating zone, it elic-
duced at sensory and postsynaptic membranes, and all-or-
its all-or-none propagated APs in the axon. When they arrive at the ter-
minals of the sensory neuron, the APs cause the release of a chemical none nerve impulses are largely confined to structures that
neurotransmitterthat induces a graded change in V, in the next neuron. are specialized for long-distance conduction, such as axons.
If the potential in the second neuron reaches threshold, an AP or a train With minor exceptions, all of the signals falling into these
of APs will be produced. Thus, graded and all-or-none potentials alter- two major categories of signal transmission-all-or-none
nate along the pathway, and the information is carried alternately by elec-
impulses and graded potential changes-are generated by
trical and chemical signals.Within each neuron the signal is carried elec-
trically, whereas between neurons the signal is carried by chemicals. the gating of specific types of membrane channels (see also
Arrows indicate the sites of chemical signal transmission. Chapters 5 and 7).
As it travels through the nervous system, information is
transformed over and over again. Sometimes it is carried by
graded changes in V,,,,which may be translated into all-or-
propagated regeneratively in this part of a sensory neuron. none, actively conducted APs. At synapses, the electrically
Just as electrical signals decay when they are carried encoded signals are changed into chemical signals in the
through a lifeless electrical conductor away from a source, form of neurotransmitter molecules, which carry the infor-
these receptor potentials progressively decay with distance mation from one cell to the pext. The chemical signal is
C O M M U N I C A T I O N A L O N G A N D BETWEEN N E U R O N S 165
...............................................................................
then reconverted into an electrical signal in the postsynap- a neuron's APs are shaped by a combination of its physical
tic neuron. The interconversions between electrical and properties and the nature of its voltage-gated ion channels.
chemical methods of conducting signals along a chain of
neurons change the character of a signal as it is transferred Passive Spread of Electrical Signals
through the nervous system. Modifications in synaptic con- The passive spread of changes in Vm occurs in all neurons.
nections are thought to underlie such complicated behav- The resistance and capacitance of the plasma membrane
ioral changes as learning. Our knowledge about synapses govern how potentials and currents spread within a cell. In
and how they can influence neuronal communication a hypothetical spherical cell, potentials would spread uni-
within the body is growing very rapidly as more neuro- formly with minimal decay, because the electrical resistance
transmitters are identified and the broad range of their ac- of the saline cytosol would be much lower than the electri-
tivities is better understood. cal resistance of the cell membrane (which depends on the
number of open ion channels-see Chapter 5). Conse-
quently, a current injected into a spherical cell would spread
TRANSMISSION OF INFORMATION out and pass through the membrane with relatively uni-
WITHIN A SINGLE NEURON form density over the entire cell surface. Neurons, however,
Information spreads through a neuron, away from its point have more complicated shapes, so the spread of a change in
of origin, through the interaction of two basic mechanisms: Vm is more complex. Many neurons have long processes
passive electrotonic conduction and active regenerativeAPs. that conduct signals over large distances. If current is in-
Electrotonic conduction takes place in all neurons, whereas jected at a point on the membrane of a long, thin, cylindri-
only those neurons with functional voltage-gated ion chan- cal region (e.g., in an axon, a dendrite, or a muscle fiber),
nels (see Chapter 5) can carry APs. Electrotonic conduction an electrical signal can spread away from that point be-
depends on the physical properties of the neuron. In contrast, cause cells possess cable properties (Figure 6-2). The cable
&--++--. P
nerve or muscle f ~ b ewr ~ t hIncreased dlstance along
t current lnject~on The
the f ~ b e rfrom the p o ~ n of
length constant, A, IS deflned as the dtstance over
w h ~ c hthe potent~alfalls by 1 - (lle), a reduct~onof
t Injedlon (V,)
63%from ~ t slnlt~alvalue at the p o ~ nof
Cf
I poten
I
Propagation of Action Potentials that are propagated along axons without any decrement. In
Chapter 5, we considered the events that produce an AP at
Nerve cells typically have long axons (see, e.g., Figure 6-11, a single location in an excitable cell. In order for APs to
so they can carry information acquired in one part of the carry information, these events must occur over and over
body to other, often distant, parts of the body by the prop- again along an axon. Basically, the signal is propagated
agation of APs. However, some neurons are sufficiently when each activated patch of membrane excites neighbor-
small that they accomplish many -even all-of their nor- ing patches. Typically, the change in Vm that occurs in an
mal electrical functions without the aid of APs. In fact, AP is about five times as great as the threshold depolariza-
many such cells are incapable of producing APs and are tion, and a patch of axonal membrane that is producing an
therefore referred to as nonspiking neurons. Their graded AP is able to excite the quiescent patch ahead of it, thereby
signals are conducted electrotonically to the axon terminals causing the AP to propagate along the axon. The difference
without the aid of all-or-none impulses. In these nonspik- between the size of the depolarization during an AP and
ing, local-circuit neurons, the amplitude of the signals is at- that of the threshold is known as the safety factor of the
tenuated as they spread through the cell, but the signals are neuron.
still large enough at the terminals to modulate the release The action potential is produced by two classes of
of a neurotransmitter. Local-circuit neurons are broadly ion channels, one selective for Na+ and the other selective
distributed in the animal kingdom. For example, they can for K+; these channels open and close with temporal pre-
be found in such widely different locations as the vertebrate cision to produce a transient reversal of the membrane
retina and other parts of the vertebrate central nervous potential (see Chapter 5). This reversal in the polarity of
system, in the barnacle eye, in the insect central nervous sys- Vmcan travel along the axon at high speed (e.g., as high as
tem, and in the crustacean stomatogastric ganglion. Local- 120 m . s-l in some large mammalian axons). At the initi-
circuit neurons are seldom more than a very few millime- ation of an AP, voltage-gated Na+-selective channels open,
ters in overall length, and they are generally characterized increasing the permeability of the plasma membrane to
by a high specific membrane resistance, which contributes Na+. When the Na+ channels are open, Na+ ions carry a
to a large length constant and, hence, to the efficient and large, but transient, current into the underlying region of
relatively undecremented electrotonic spread of signals. the axon. This inward current spreads longitudinally along
More typically, however, communication between dif- the axon and then leaks out across the membrane to com-
ferent parts of the nervous system depends critically on the plete the circuit of current flow. The longitudinal electro-
propagation of APs along the axons of neurons, because tonic spread of current depends on the cable properties of
the distances are too great for the electrotonic spread of sig- the axon described earlier. Thus, the influx of Na+ that pro-
nals to be effective. Similar propagation also takes place in duces the upstroke of the AP (Figure 6-4A) supplies a cur-
many muscle cells. As discussed in Chapter 5, APs are large rent that spreads longitudinally, both forward and poten-
voltage changes across the neuronal membrane that have tially backward as well, from the point of origin within the
an invariant shape when plotted as a function of time and axon (Figure 6-4B). The elecpotonic spread of current
168 P H Y S I O L O G I C A L PROCESSES
...............................................................................
Figure 6-4 Electrotonicspread of current carries an action potential from
one region of an axon t o the next. (A) As an AP travels along an axon,
electrodes placed along the axon record the same-sized change in V,,, at
successively later times. (B) An AP propagates along an axon as current
from (1) an active patch of membrane (dark shading) spreads electroton-
ically t o (2) a neighboring inactive patch o f membrane, depolar~zingit
(light shading) and bringing it t o threshold. Then current across (3) the
newly active patch of membrane spreads t o (4) a new inactive patch of
membrane just ahead, depolarizing it and bringing it t o threshold. This
process is repeated over and over again along the axon. In this figure, the
AP is traveling from left t o right. A patch o f membrane that has just re-
turned t o its polarized state is transiently incapable of becoming acti-
vated again, so the AP is propagated in only one direction. (See text for
further discussion.)
I
Axon
Time (ms)
away from active patches of membrane is responsible for of membrane partially depolarizes (compare Figures 5-16
and 6-4A). In fact, as an undergraduate in 1937, Alan
When positively charged ions enter the patch of axonal Hodgkin confirmed the hypothesis that the inactive mem-
membrane that is immediately ahead of the AP, that patch brane ahead of an AP becomes depolarized by electrotoni-
cally conducted current. His experiment is illustrated in primarily on open K+ channels, the outward current is car-
Figure 6-5. Electrical current can flow in a circuit only if ried primarily by K+.)
the circuit is complete, and this physical law must be As the membrane ahead of the impulse becomes depo-
obeyed in axons as well. The current that flows longitudi- larized by local circuit current, voltage-gated Na+ channels
nally within an axon-that is, in the direction of impulse in that membrane open (i.e., gNaincreases),initiating an AP
propagation (to the right in Figure 6-4B)-must flow out in this new patch of membrane. The newly excited region
of the axon-across unexcited parts of the membrane that then generates a local-circuit current that delplarizes, and
lie ahead of the region of Na+ influx-and then back into thereby excites, parts of the axon ahead of it. Thus, local-
the active region of the axon to complete the circuit. (Be- circuit current from each excited region depolarizes and ex-
cause the conductance of the resting membrane depends cites the region immediately ahead of it. In this manner, the
signal is continuously boosted and maintained at full
strength as it travels along the axon. Note how different
this method of signal transmission is from the decremented
electrotonic conduction in passive transmission. The
amount of depolarization required to bring an inactive
membrane to threshold is about 20 mV, whereas the total
depolarization during an AP is typically about 100 mV.
Thus, an AP produces an approximately fivefold "boost"
of the electrotonic signal.
Some of the current that enters an axon at the excited
region spreads backward within the axon-that is, in the
Cold
direction from which the impulse originated. However,
block under normal conditions, this backward-moving current
cannot excite the membrane and produce a backward-
traveling AP,because the membrane just behind a region of
advancing excitation is in a refractory state (see Chapter 5).
The Na+ channels in that region are inactivated, and the K+
channels are still open; so current is carried out of the cell
as an efflux of K+, preventing depolarization in that region.
The delayed activation of K+ channels is also important be-
Nerve cause it speeds up the repolarization of the membrane,
E priming it for future APs.
c---+
-'id-+
Sooty paper
moves
Figure 6-6 Hermann von Helmholtz measured the t ~ m efrom st~mulusto posltlon S,, the paper having been al~gnedso that the t ~ m eof the sec-
contract~onIn a frog nerve-muscle preparation to determ~neconduct~on ond st~mulusexactly co~nc~ded wth the t ~ m eofthef~rstst~mulusThe trac-
veloc~tyalong a nerve Electrodes to stimulate the nerve weref~rstplaced lngs made by the lever showed the change In latency, At, when the loca-
at posltlon S,, and contract~onof the muscle moved a lever that tlon of the electrodes was changed The conduct~onveloc~tycan be
scratched a record on a rap~dlymovng sheet of sootypaper (Not~ce that calculated from the d~fferenceIn latency of the muscle twitch evoked by
t ~ m eIS plotted as the paper moves ) Then the electrodes were moved to stimulat~ngthe nerve at the two d~fferentlocat~ons
TABLE 6-1 A
Nerve trunk Crushed
The diameter o f frog axons and the presence o r absence
o f myelination control the conduction velocity w
Axon diameter Conduction velocity kel
Fiber type (~m) (m . S-') Stimulus
Myelinated fibers
Aa 18 5 42
B
AP 14.0 25
AY 11 0 17
€3 Approx 3.0 4.2
Unmyelinsted fibers
C 25 0 4-0 5
by Erlanger and Gasser, 1937. Adapted from
Source Flber class~f~catlon
Davson, 1964
Oligodendrocyte
Node of
Ranvier
Axon
Myelin sheath
k m p o s e d of layered
glial cell membrane
Terminal
/ Plasma membrane of axon
loops of
Schwann ce
Myelin
Figure 6-8 Myellnated axons are wrapped by supporting cells that leave as 50 Internodes on several adjacent axons Thls schematic lllustratlon
short segments of axonal membrane exposed at the nodes of Ranvler does not show how tlghtly the layers of myelln are wrapped around the
(A) Ashort segment of axon, called a node of Ranvler and located be- axon (B)Electron micrograph of a node of Ranvler In a splnal root of a
tween two myel~n-wrappedInternodes, sexp posed to extracellularflu~ds young rat In these nerves, a segment of axonal membrane about 2 p m
Only the membrane at such nodes becomes exclted dur~ngsaltatory long IS exposed to extracellularflu~d[Part B courtesy of Mark Elllsman ]
conduction Aslngle ollgodendrocyte cell can provlde myelln for as many
172 PHYSIOLOGICAL PROCESSES
.........................................
SPOTLIGHT 6-1 FacingpageThe activity of axons in a nerve bundle can be recorded with
extracellular electrodes. Stippled areas in each diagram show the loca-
tion of the depolarized membrane at five different times, from t, to t,.
EXTRACELLULAR The records show what you would see-for example, on an oscillo-
scope-for each method of recording. Numbers on the records corre-
SIGNS OF IMPULSE spond to the five different times. In all cases, the oscilloscope signal is
B I I I Injured section
Time
myelin layer is very thick. This reduction in C, means that era1 nerves and ~ l i ~ o d e n d r o c y tin
e sthe central nervous sys-
less capacitative current is required to change Vm, so more tem. Between nodes of Ranvier, the sheath is so close to the
charge can flow down the axon to depolarize the next seg- axon membrane that it nearly eliminates the extracellular
ment. The changes in resistance and capacitance greatly in- space surrounding the axon membrane. Moreover, the in-
crease the length constant, A, of the axonal membrane that ternodal axon membrane has been found to lack voltage-
is covered by myelin, thus enhancing the efficiency with gated Na+ channels. Thus, when a local-circuit current
which longitudinal current spreads. However, this insula- flows in advance of the AP, it exits the axon almost exclu-
tion would not have this effect if it completely covered the sively through the nodes of Ranvier. As noted earlier, very
axon, because the electrot~nicall~ conducted current would little current is expended in discharging membrane capac-
eventually decrease to zero as a function of distance. In- itance along the internodes, because of the low capacitance
stead, the length of the myelinated segments are typically of the thick myelin sheath. An AP that is initiated at one
about 100 times the external diameter of the axon, ranging node electrotonicallydepolarizes the membrane at the next
from 200 pm to 2 mm long, and the segments are in- node; thus, in myelinated axons, APs do not propagate
terrupted by short, unmyelinated gaps called nodes of continuously along the axonal membrane, as they do in
Ranvier, at which about 10 pm of the excitable axon is nonmyelinated nerve fibers. Instead, APs are produced only
exposed to the extracellular fluid (Figure 6-8B). The seg- in the small areas of the membrane exposed at the nodes of
ments of axon that lie under the myelin wrapping are called Ranvier. The result is saltatory conduction, a series of dis-
internodes. continuous and regenerative depolarizations that take place
In the course of development, myelin is laid down only at the nodes of Ranvier, as illustrated in Figure 6-9.
around the axons of peripheral and central tracts in verte- The velocity of signal transmission is greatly enhanced be-
brates by two kinds of glial cells: Schwann cells in periph- cause the electrotonic spread of local circuit current occurs
Direction of propagation Figure 6-9 In saltatory conduction, the action
~ o t e n t i ailu m ~ from
s node to node. (A) Cur-
rent spreads longltudlnally between nodes
The large red arrows lndlcate Na+ Influx
through aalvated Na+ channels that are lo-
cated at the nodes The smaller red arrow rep-
resents the later outfux of Kt through act!-
vated K+ channels (B) Dots indlcate the value
of V , at a single instant at each node shown in
part^. ~ t s i t Ie, thernernbrane IS in the falling
-
phase of an AP; at site 2, the membrane is in
the rlslng phase At sltes 3,4, and 5, the mem-
1-1 1-1 I^\ 1-6
I
- ,' \\
*
I \
--d * - - 4
t - 4 '
+-I\
* 4
i \\
*-,
brane IS In success~vel~ earher phases of an AP
5 4 3 2 1
rapidly over internodal segments. The conduction velocity neurotransmitters have been identified in the wide range of
of myelinated fibers varies from a few millimeters per sec- animals studied, more are being discovered all the time,
ond to more than 100 m . s-l, in contrast with unmyeli- and we now know that their modes of action vary greatly.
nated fibers of similar diameter, which conduct at a fraction Initially, neurotransmitters were thought to act only by
of a meter per second (see Table 6-1). causing the voltage in a postsynaptic cell to change, either
The evolution of saltatory conduction and the result- by hyperpolarizing the cell or by depolarizing it. Howevel;
ing higher speed of AP propagation was probably crucial neurotransmitters can also increase or decrease the num-
for the successful coordination of activity in the large ber of ion channels inserted into the membrane of the post-
muscles of vertebrates. Myelination allows APs to travel synaptic cell, alter the excitability of the postsynaptic cell
rapidly in many axons within a compact nerve trunk. The by changing the rate at which ion channels open and close,
importance of the specialized insulation provided by or modify their sensitivity to activating signals. The dis-
myelin for coordinating neuronal information is particu- covery of these varied modes of action has dramatically
larly evident in such demyelinating diseases as multiple broadened our understanding of the role that synapses
sclerosis. In this disease, the myelin sheath is reduced or play in neuronal communication.
eliminated along some axons making the velocity of Synaptic transmission was a subject of controversy for
neuronal transmission highly variable among neurons, a very long time. Early in the twentieth century, the great
which severely compromises the control of coordinated histologist Santiago Ramon y Cajal used the light micro-
movement. scope and a silver-based staining technique developed by
neuroanatomist Camillo Golgi to show that neurons stain
as discrete units. In spite of this observation, many anato-
TRANSMISSION OF INFORMATION mists continued to believe that the nervous system was a
BETWEEN NEURONS: SYNAPSES continuous reticulum, rather than a set of morphologi-
All information processing done by neurons depends on cally separate nerve cells. It was not until electron mi-
the transmission of signals from one neuron to another, croscopy was developed in the 1940s that unequivocal ev-
which is accomplished at structures called synapses. At idence was obtained supporting the notions that neurons
electrical synapses, the presynaptic neuron is electrically are indeed separate from one another and that particular
coupled to the postsynaptic neuron by particular proteins regions of neurons are specialized for communication
within the membranes. Transmission across electrical between cells.
synapses proceeds very much like signal transmission However, in 1897, long before the ultrastructural basis
along a single axon. However, electrical synapses are rel- of neuron-neuron interactions was determined, the func-
atively rare. Most signaling between neurons takes place tional junction between two neurons was given the name
at chemical synapses. At a chemical synapse, APs in the synapse (from the Greek, meaning "to clasp") by Sir
presynaptic neuron cause the release of neurotransmitter Charles Sherrington, who is widely regarded as the founder
molecules that diffuse across a narrow space (about 20 nm of modern neurophysiology. It was his conclusion that
wide), called the synaptic cleft, that separates the mem- ". . . the neurone itself is visibly a continuum from end to
branes of pre- and postsynaptic neurons. As recently as the end, but continuity fails to be demonstrable where neurone
1970s, only a handful of chemicals were known to be meets neurone-at the synapse. There a different kind of
synaptic transmitters. All of them were thought to act in transmission may occur" (Sherrington, 1906). Although
a similar fashion, in accord with the results obtained in Sherrington had no direct information about the mi-
studies of transmission at the synapses, called neuromus- crostructure or microphysiology of these specialized re-
cular junctions (NMJs), between motor neurons and the gions of interaction between excitable cells, he had extra-
skeletal muscles that they control. Today, more than 50 ordinary insight, the sources of which were his cleverly
designed experiments on the spinal reflexes of animals, of an AP (the ratio of the change in V,,, during an AP to
most of them mammals. Among other things, he deduced the change in Vm required to bring a cell to threshold)
that some synapses are excitatory, increasing the probabil- is typically about 5, the attenuation of the change in
ity that APs will arise in the postsynaptic cell, and that oth- V,,, from one cell to the next must be no greater than the
ers are inhibitory, reducing the probability of APs in the safety factor if the depolarization of the postsynaptic cell
postsynaptic cell. is to reach threshold and initiate an impulse. Therefore, a
In this section, we start by considering synaptic trans- single presynaptic action potential might be unable to
mission across electrical synapses, which is similar to signal provide enough local circuit current across an electrical
conduction along axons. We then turn to the topic of chem- synapse to elicit an action potential in the postsynaptic
ical synapses, dealing first with transmission at the neuro- cell, which may be one evolutionary reason why electrical
muscular junction and then with other, more recently dis- synapses are less common than chemical synapses. How-
covered types of chemical synapses. ever, the fact that electrical synapses conduct signals
much more rapidly than do chemical synapses gives them
Synaptic Structure and Function: definite advantages where rapid signal transmission is
Electrical Synapses important.
Electrical synapses transfer information between cells by di- Electrical transmission between excitable cells was first
rect ionic coupling. At an electrical synapse, the plasma discovered in 1959 by Edwin J. Furshpan and David D.
membranes of the pre- and postsynaptic cells are in close Potter, who were studying the nervous system of crayfish at
apposition and are coupled by protein structures called gap the time. They found that an electrical synapse between the
junctions (Figure 6-10A) through which electrical current crayfish lateral giant nerve fiber and a large motor axon has
can flow directly from one cell into the other (Figure 6-10B; the unusual property of passing current preferentially in
see also Chapter 4).Because current travels across gap junc- one direction (see Figure 6-10D). Since their early work,
tions, an electrical signal in the presynaptic cell produces a electrical transmission has been discovered between cells in
similar, although somewhat attenuated, signal in the post- the vertebrate central nervous system and in the vertebrate
synaptic cell by simple electrical conduction through the retina, between smooth muscle fibers, between cardiac
junction (Figure 6-10C).Thus, at an electrical synapse, the muscle fibers, between receptor cells, and between axons.
transfer of information occurs by purely electrical means, The rapidity
- .
with which current crosses electrical synapses
without the intervention of a chemical transmitter, and a makes this means of information transfer particularly ef-
key feature of electrical synaptic transmission is its rapidity. fective in the synchronization of electrical activity within a
As we will soon see, signal transmission across chemical group of cells. It is also effective for rapidly transmitting in-
synapses is always slower than purely electrical signal formation across a series of cell-cell junctions-for ex-
transmission. ample, in the giant nerve fibers of the earthworm, which are
Electrical transmission can be illustrated experimen- composed of many segmental axons connected in series
tally by injecting current into one cell and measuring along the worm's body, and in the myocardium of the ver-
the effect in a connected cell (see Figure 6-10C). A sub- tebrate heart, in which signals are passed between muscle
threshold current pulse injected into cell A elicits a tran- cells.
sient change in the membrane potential of that cell. If a At some synapses, transmission is both electrical and
significant fraction of the current injected into cell A chemical. Such combined synapses were first identified in
spreads through gap junctions into cell B, it will cause a cells of the avian ciliary ganglion, and they have also been
detectable change in the V,,, of cell B as well. Because found in a circuit controlling the fish escape response, in
there is a potential drop as the current crosses the gap synapses made by some neurons onto spinal interneurons
junctions, the potential change recorded across the mem- of the lamprey, and in synapses onto frog spinal motor
brane of cell B will always be less than that recorded in neurons. However, as interesting as they are, combined
cell A. The gap junctions through which current flows
- -
synapses are unusual phenomena.
from one cell to another are generally, but not always,
symmetrical in resistance to the passage of current-that Synaptic Structure and Function:
is, current generally meets the same resistance as it passes Chemical Synapses
in either direction between the two cells. However, at A common mode of synaptic transmission is known as fast
some specialized synapses, the transfer of current between chemical synaptic transmission, which is found at many
the two coupled cells occurs readily in one direction, but synapses in the central nervous system and at the neuro-
not in the other (Figure 6-10D). Such junctions are said to muscular junction. (Although this transmission is called
be rectifying. "fast," it is in fact considerably slower than transmission
The transmission of an AP through an electrical across electrical synapses.) At neuromuscular junctions,
synapse is basically no different from propagation within the neurotransmitter acetylcholine (ACh) is stored in
one cell, because both phenomena depend on the passive membrane-enclosed vesicles and is secreted by exocytosis
spread of local circuit current beyond the AP to depolar- into the extracellular fluid separating the neuron and the
ize and excite the region ahead. Because the safety factor muscle. The sequence of events at these nerve terminals is
176 P H Y S I O L O G I C A L PROCESSES
......................................................................
A I /
Current
passes between cells
through gap junctions
D
Ant~dromic
Current source
Pre.
50 mV 25 mV
\
/
Receptor ~ostsina~tic
protein membrane
Figure 6-11 In fast chemical synaptic transmission, signals in the pre- synaptic cleft by exocytosis. (C) Neurotransmitter molecules diffuse
and postsynapticcellsare linked by chemical neurotransmitters. The pre- across the synaptic cleft, driven by their concentration gradient, and bind
and postsynaptic cells are not electrically coupled, and there is no direct to receptor proteins in the postsynaptic membrane, opening ligand-
flow of current between them. Ionic current flows across the postsynap- gated ion channels. In this case, Na' flows through the open channels
tic membrane only when ligand-gated ion channels are open in the post- into the presynaptic cell. The vesicle membrane remains fused with the
synaptic membrane. (A) At rest, transmitter molecules are packaged into membrane of the terminal, but it moves to the sides of the terminal.
membrane-bounded vesicles contained In the axon terminals. (B) When (D) Transmitter molecules are removed from the cleft, the postsynaptic
an AP enters the presynaptic terminal, it causes voltage-gated Ca2+chan- ion channels close, and the membrane that was added to the presynap-
nels in the membrane to open, allowing Ca2+ions to flow into the termi- tic terminal when the synaptic vesicles fused is eventually recycled into
nal. The increase in intracellular free Ca2+causes synapticvesicles to fuse the terminal (small arrows) and may be reused for more vesicles.
with the presynaptic membrane, releasing neurotransmitter into the
A B
Fast chemical Slow chemical
transmission transmission
Large
vesicle
Receptor
Figure 6-12 Fast chem~calsynaptic transmlsslon and slow chem~cal These transmitters are stored in large, distinctive vesicles and are re-
synapt~ctransm~sslonact through different postsynaptlcmechan~sms(A) leased from sites that lack morphological specialization and that are lo-
In fast chem~caltransm~ss~on, neurotransm~tters are synthes~zed In the ter- cated away from the sites from which the fast neurotransmitters are re-
m~nalsand stored In small, clearves~clesThese transmltters are typically leased. In the postsynaptic cell, these neurotransmitters typically act
small molecules The ves~clesare located near the plasma membrane, indirectly through G protein-linked receptors to modify channels and
and transmltters are released by exocytos~sInto the synapt~ccleft through other intracellular processes. Single neurons may produce both kinds
spec~al~zed sltes on the membrane After they are released, these neu- of transmission, and a single neurotransmitter may affect postsyn-
rotransmltters act on I~gand-gatedchannels In the postsynapt~cmem- aptic neurons both by means of ligand-gated channels and by means of
brane (B) In slow synaptlc transm~ss~on, the transmltters are typ~cally G-protein-coupled receptors.
larger molecules-for example, pept~desconta~nlngmany ammo ac~ds
which they are transported to the nerve terminal. Vesicles postsynaptic current produced by the neurotransmitter
that mediate slow synaptic transmission may release their causes a change in the membrane potential of the post-
transmitter molecules at many sites in the presynaptic ter- synaptic cell. If the sum of the potential changes caused by
minal and usually affect the postsynaptic cell, not through many such synaptic events is sufficient to exceed the thresh-
ligand-gated channels but by altering the levels of intracel- old potential in the postsynaptic cell, an AP will be initiated
lular second messengers through intermediate molecules in the postsynaptic cell.
called G proteins. Physiological and anatomical evidence In fact, the currents that are generated in the post-
indicates that single presynaptic neurons may participate in synaptic cell may either increase or decrease the probabil-
both kinds of neurotransmission. ity that APs will occur in that cell; that is, synaptic effects
The release of neurotransmitter into the synaptic cleft can be either excitatory or inhibitory. What makes a synap-
is controlled by mechanisms that are common to both fast tic signal one or the other is examined later in this chapter.
and slow synaptic transmission. When an AP arrives at the
axon terminals, it activates voltage-gated Ca2+channels in
the membrane of the terminals, allowing Ca2+to enter the
terminal (seeFigure 6-11B). The increased concentration of
Ca2+inside the terminal initiates the exocytosis of vesicles
containing the transmitter substance, dumping neuro-
transmitter molecules into the synaptic cleft where they dif-
fuse away from the presynaptic terminal. In fast synaptic
transmission, neurotransmitter-containing synaptic vesicles
fuse with the plasma membrane at specialized sites called
active zones. After crossing the synaptic cleft, some neuro-
transmitter molecules bind to receptor molecules in the Fast Chemical Synapses
postsynaptic membrane. When transmitter molecules bind The most extensive studies of synaptic transmission have
to these receptor molecules, they modify ionic current been done on fast chemical transmission at the neurbmus-
through channels that are associated with the receptor mol- cular junctions (also called motor terminals or motor end-
ecules, allowing permeant ions to carry a postsynaptic cur- plates) of vertebrate skeletal muscle, where acetylcholine
rent driven by electrochemical gradients. In slow transmis- has been shown to be the neurotransmitter. We will use the
sion, the neurotransmitter affects the postsynaptic cell neuromuscular junction as our primary example, because
through G-protein intermediates to modify activities of in- it is so well studied. It is a good example because fast chem-
tracellular second messengers that then influence ion chan- ical synaptic transmission between neurons within the cen-
nels or other intracellular processes (see Figure 6-12).The tral nervous system closely resembles transmission at the
C O M M U N I C A T I O N ALONG A N D BETWEEN NEURONS 179
..............................................................................
neuromuscular junction, although in many cases the trans- are thousands of vesicles, each about 50 nm in diameter, in
mitters are different. one presynaptic terminal. For example, the branches of the
nerve terminal innervating a single frog muscle fiber typically
Structural features contain a total of about 10"synaptic vesicles. When the vesi-
The frog motor endplate (Figure 6-13) includes structural cles fuse with the plasma membrane and release transmitter
specializations of the presynaptic terminal, of the postsy- molecules into the synaptic cleft, the transmitter molecules
naptic membrane, and of associated Schwann cells. The reach the postsynaptic membrane by diffusing down their
axon of the presynaptic motor neuron terminal bifurcates, concentration gradient. The cleft itself is filled with a mu-
and the branches, each of which is approximately 2 p m in copolysaccharide that "glues" together the pre- and postsy-
diameter, lie in a longitudinal depression along the surface of naptic membranes, both of which usually show some degree
the muscle fiber. The muscle membrane lining the depression of thickening at the synapse. The vesicular membrane that
is thrown into transverse junctional folds at intervals of 1to fused with the plasma membrane of the terminal is taken up
2 pm. Directly above these folds within the nerve terminal into the terminal and may be recycled (see Figure 6-1lD).
are the active zones-transverse regons of slight thickening When acetylcholine (ACh)is released into the synaptic
in the presynaptic membrane above which are clustered cleft, it can bind to ACh-specific receptor molecules in
many synaptic vesicles. The vesicles are released along the ac- the postsynaptic membrane of the endplate, causing ion
tive zones by the process of exocytosis (Figure 6-14).There channels that are selective for Naf and K+ to open briefly.
Mvelinated axons
Nerve terminals
,Synaptic
vesicles
,Synaptic
cleft
Myelin
B
Schwann cell
'
\ / Junctional
fold
Muscle rnrn
fiber
0 5 10
T ~ m e(ms)
These results suggest that curare interferes with synap- an unattenuated manner because it is regenerative, the
tic transmission by blocking endplate potentials in propor- synaptic potential spreads passively and thus decays with
tion to its concentration. If the concentration of curare is distance. In experiments like this one, curare enabled phys-
sufficient to reduce the size of the synaptic potential in the iologists to distinguish among the elements of the synaptic
muscle to just below threshold, the AP is eliminated, and response in vertebrate muscle fibers.
the synaptic potential is revealed without a superimposed
AP (see Figure 6-15C). If the recording electrode is now Synaptic currents
reinserted into the muscle fiber a number of times at pro- As described in Chapter 5 , a change in membrane perme-
gressively greater distances from the motor endplate, the ability for one or more species of ion (i.e., opening or clos-
amplitude of the measured postsynaptic potential drops ap- ing a population of membrane channels that selectively
proximately exponentially with distance from the endplate passes those ions) typically shifts the membrane potential
(Figure 6-16). In contrast with the AP, which propagates in toward a new level. This change in V, occurs because when
182 P H Y S I O L O G I C A L PROCESSES
...............................................................................
SPOTLIGHT 6-3 texln from the tlger snake also lnhlblts transmitter release, caus-
Ing lethal paralysrs.The evolut~onof these toxlns has made them
PHARMACOLOGICAL hlgh~yeffectwe for lncapacltatlng a vlctlm (I e , only very small
amounts are needed), and they must be handled wlth great cau-
AGENTS USEFUL IN tion In the laboratory
SYNAPTIC STUDIES
Postsynaptic Receptor Toxins
Studies of axonal and synaptic transmission have been greatly Agonists and antagonists for receptor subtypes have con-
aided by the discovery and application of natural toxins-from tributed importantly to defining the role of these receptors in
animals, plants, or fungi-that selectively interfere with, or par- neuronal processing. y-Aminobutyric acid (GABA), a largely
tially mimic, certain steps in the process of transmission. Toxins inhibitory neurotransmitter, has been studied extensively
have been found that interact with ion channels, with receptors, with the use of a pair of chemicals, one an agonist of GABA and
and with enzymes important for nervous system function. Some the other an antagonist. The agonist, muscimol, is derived from
of the commonly used agents that have been useful in studies of the mushroom Amanita muscaria. It specifically acivates
synaptic transmission are described here. GABA, type CI- channels. Bicuculline, produced from the plant
Dicentra cucullaria, is a competitive antagonist of the same
Channel Toxins channel.
Several toxins are specific for particular types of ion channels. A huge collection of reagents exists for ACh receptors. Mus-
Tetrodotoxin (TTX) from the puffer fish (Sphoeroidessp.) binds carine and other agents, including pilocarpine, activate mus-
to a site on voltage-gated Na+ channels and blocks Na+ current carinic ACh receptors. In vertebrates, muscarinic ACh receptors
across the membrane. Similarly, saxitoxin (STX), derived from di- are most prevalent in the visceral tissues that are innervated by
noflagellates, blocks voltage-gated Na+ channels, although by the cholinergic axons of the parasympathetic system. Atropine
a slightly different mechanism. Potassium ion channels can be (belladonna) is a plant-derived alkaloid that blocks muscarinic
blocked by several agents. For example, tetraethylammonium synaptic transmission.
(TEA), a synthetic organic compound, blocks most types of K+ Nicotine, another plant alkaloid, and certain other agents,
channels from either inside or outside the membrane, and such as carbachol, act as agonists of nicotinic ACh receptors.
4-amino pyridine blocks several types. Calcium ion channels can D-Tubocurarineis the active principle of curare, the South Amer-
be blocked by any of several w-conotoxins derived from the ican blow-dart poison, made from the plant Chondodendron to-
piscivorous (fish-eating) cone snail (Conus geographus). The mentosum. This molecule blocks transmission postsynaptically
various subtypes of this toxin block different classes of Ca2+ by competing with ACh for the ACh-binding site of the nicotinic
channels. receptors. It binds competitively to these sites without opening
Glutamate-gated channel toxins have proved invaluable in the channels, and it thereby interferes with the generation of a
distinguishing among the variety of channel types. Kainic acid, postsynaptic current. Similarly, a-Bungarotoxin (a-BuTX) is iso-
from a red alga (Digeneasimplex), is an effective agonist for one lated from the venom of the krait, a member of the cobra fam-
subtype of glutamate receptors. Quisqualic acid, derived from ily. This protein molecule binds irreversibly and with very high
seeds of the plant Quisqualis indica, is a second potent agonist specificity to nicotinic ACh receptors. With the use of radioac-
that is selective for another subtype. One important antagonist tively labeled a-BuTX, it has been possible to determine the
is conatokins, from cone snails, which is a noncompet~tivean- number of ACh receptors present in a membrane as well as to
tagonist of a third class of glutamate receptors, called NMDAre- isolate and purify the receptor protein.
ceptors for N-methyl-D-aspartate,which activates them. Eserine (physostigmine) is an anticholinesterase; that is, it
blocks the action of acetylcholinesterase.Use of this alkaloid has
Presynaptic Toxins enabled physiologists to measure the amount of ACh released
Several toxins act on presynapticterminals to inhibit transmitter at a synapse, by preventing the rapid enzymatic destruction of
release. /?-Bungarotoxin, derived from cobra venom, inhibits the transmitter molecules. Partial doses accentuate the post-
transmitter release by permeabilizing the nerve terminal. No- synaptic potential at cholinergic synapses.
channels open they allow a flow of ions that transfers of ions through the membrane.) The direction and intensity
charge from one side of the membrane to the other, which of the synaptic current, which are controlled by the size of
in turn causes the measured transmembrane voltage to the conductance through the open channels and by the elec-
change. In chemical synaptic transmission, postsynaptic trochemical driving force and charge on the permeant ions,
channels in the membrane open when neurotransmitter determine the polarity and the amplitude of the postsynap-
binds to receptor proteins, and a synaptic current can then tic potential. Because neurotransmitters activate channels
flow through these newly opened postsynaptic channels. (In with selective ionic permeabilities, they confer specificity on
some cases, postsynaptic channels close, reducing the flow synaptic signal transmission by allowing only certain ionic
C O M M U N I C A T I O N ALONG A N D BETWEEN NEURONS 183
...............................................................................
species to cross the postsynaptic membrane in response to romuscular junction consists of an influx of Na+ that is
particular neurotransmitters. partly canceled by a simultaneous, and somewhat smaller;
The ionic currents that produce postsynaptic potentials efflux of Kf. At this synapse, both Naf and K+ ions pass
can be recorded by voltage-clamping the postsynaptic through the very same postsynaptic ACh-activated chan-
membrane, thus holding the postsynaptic potential con- nels, indicating that these channels have a broader ion se-
stant (see Spotlight 5-3). In a nerve-muscle preparation, this lectivity than do the highly selective, voltage-gated Na+ and
procedure must be carried out close to the motor endplate K+ channels that underlie APs (see Figure 5-26).
(Figure 6-17A). The motor nerve (the presynaptic element) Synaptic currents last a considerably shorter time than
is stimulated while V, of the postsynaptic membrane is d o synaptic potentials (see Figure 6-17B). Acetylcholine-
voltage-clamped at some predetermined value. The release activated channels open only briefly, because the transmit-
of transmitter by the presynaptic nerve ending is quickly ter at the neuromuscular junction is rapidly removed from
followed by a synaptic current (Figure 6-17B) that is pro- the cleft by enzymatic destruction, after which the channels
duced when ions flow down their electrochemical gradients close and the synaptic current ceases to flow. A postsynap-
through open channels in the postsynaptic membrane. tic potential lasts longer than the synaptic current because
The ions responsible for carrying the synaptic current its time course depends on the time constant of the mem-
at particular synapses have been identified through exper- brane, as well as on the duration of the synaptic current.
iments in which the extracellular concentrations of specific
ions were changed and the resulting effect on the synaptic Reversal potential At every fast chemical synapse; one (or
current was measured. Such measurements demonstrated more) species of ions carries current across the postsynap-
that the depolarizing synaptic current at the vertebrate neu- tic membrane, and the change in V, caused by this current
determines whether the synapse is excitatory or inhibitory.
Measuring the properties of the synaptic current provides
A an experimenter with clues to the identity of the ions that
Stimulating wire
carry the synaptic current. These measurements are made
7 Recordlng
electrode - V,,
by injecting current into the postsynaptic cell to set the
membrane potential at different values and then observing
the sign and amplitude of the postsynaptic potential pro-
duced by synaptic inputs (Figure 6-18A and B). The ampli-
tude and sign of the postsynaptic potential depend on the
transmembrane voltage and on the species of ion or ions
carrying the current. Remember that activation of mem-
brane channels that select for a given ionic species, X,
causes V,,, to move closer to the equilibrium potential, Ex,
-
for that ion (see Chapter 5).
electrode Consider the experiment that is illustrated in
fiber Figure 6-18 for a synapse at which only one ionic species,
X, carries the synaptic current. As the membrane potential,
V,, is shifted toward the equilibrium potential, Ex, the
B Undamped synaptic membrane driving force on X (V, - Ex), will decrease. When
V,,, = Ex, no current will flow across the membrane, even
though the channels are open, because there is no driving
force on the ions. If in the experiment V,,, is set on the other
side of Ex,current will once again flow, because V,,, - Ex
1, 1 k ...,.I-
y n a p t i c current
.:=:i
will again be nonzero, but the sign will have changed, indi-
cating that the driving force is in the opposite direction. As
a result, X will flow through the open channels in the di-
I 50 m s rection opposite to that of its previous flow, and the sign of
Motor axon stimulated the postsynaptic potential will be opposite to that of its pre-
vious value (Figure 6-18B and C). Because the direction of
Figure 6-17 Voltage-clamping the postsynaptic membrane allows the
synaptic current to be measured. (A) Setup for voltage-clamping the the ionic current and the sign of the postsynaptic potential
muscle membrane, which holds the postsynaptic potential constant while reverse as V,,, passes through Ex, Ex is called the reversal
ionic current flowing across the postsynaptic membrane through chan- potential, Ere".When synaptic channels open, the synaptic
nels opened by a neurotransmitteris recorded (see Spotlight 5-3).(6) The current causes V,,,to shift toward the Ere"of the current, no
upper record shows an endplate potential when the neuron is stimulated
matter where V,,,was set experimentally before the synapse
and the muscle is not voltage-clamped. The lower record shows a synap-
tic current when the muscle fiber is voltage-clamped under the same con- was activated. The reversal potential has proved to be a
ditions. The synaptic current decays much faster than does the endplate useful property of synaptic currents because it provides a
potential. hint about which ions carry the current. In fact, before
184 P H Y S l O L O G l C A L PROCESSES
.........................................
Figure 6-18 The synaptic reversal potentlal is measured by changing the
membrane potential and recording the postsynaptic potential. (A)
Method for determining the reversal potential (E,,) at a synapse. Steady
current is injected into the postsynaptic cell with an electrode to set V , at
different values. At each value of V ,, an endplate potential is produced
by stimulating the presynaptic nerve. The endplate potentials are
recorded with a second electrode in the postsynaptic muscle fiber. (B) At
first (bottom), V
, IS set at values that are more negative than the equilib-
rium potential, Ex, for the ions carrying the synaptic current. When V, is
set equal to Ex, no synaptic current flows, and the amplitude of the post-
synaptic potential is zero, even though the postsynaptic ion channels are
open. When V , is set at values more positive than Ex,the dr~vingforce on
the ions carrying the synaptic current is opposite to the direction of the
driving force when V , is more negative than Ex.As a result, when V, is
more positive than Ex, ionsflowthrough the synaptic channels in the op-
posite direction from thelr direction when V , is more negative than Ex,
Reversal potential and the sign of the endplate potential reverses. (C) The results of this type
of experiment can be plotted to show the amplitude of the endplate po-
- O tentials as a function of the values of Vm The line fitting the experimen-
tal points crosses the abscissa at E,". In thls case, E,ev= 0 mV.
-
T~me
potasslum currents through activated acetyl-
chollne (ACh) channels at different membrane
potentials, beglnnlng at EN, The ACh-act~vated
channels are approximately equally permeable
to Na' and K+, so the rnagnltudes of ,I and I,
depend on the driv~ngforce on each ion The rel-
atlve magn~tudesof the Na' and K+ currents are
representedby the lengths of the arrows (B) The
arnpl~tudeand tlme course of the net current
through the ACh-aa~vatedchannels are shown
-t
against tlme At Ere"for the comb~nedcurrents,
the net current through the channels IS zero
c v m = Ere"
AC h
release
in Vmthat can be produced by the activation of synaptic old. If E,,, for inhibitory channels is more negative than
channels (or indeed by the activation of any ion channels). Vre, in the postsynaptic cell, the synaptic current will cause
The reversal potential also has a special functional signifi- Vmto become more negative than V, ,hyperpolarizing the
cance at synapses, because the relation between EreVand the cell toward Ere, (see Figure 6-20A). Hyperpolarizing synap-
threshold for excitation in the postsynaptic cell determines tic currents can add to depolarizing synaptic currents, re-
how synaptic events affect the postsynaptic cell. ducing the net amount of depolarization in the postsynap-
tic cell.
Postsynaptic excitation and inhibition Any synaptic event Although all excitatory synapses generate depolarizing
that increases the probability that an AP will be initiated in postsynaptic currents, there are special cases among in-
the postsynaptic cell is called an excitatory postsynaptic hibitory synapses. For example, if Ere"for a synaptic cur-
potential (epsp);conversely, any synaptic event that reduces rent happens to be identical with Vrest(Vm- Ere" = 0), no
the probability of an AP in the postsynaptic cell is a n net synaptic current will flow even if postsynaptic channels
inhibitory postsynaptic potential (ipsp). If the reversal po- open. The net current will be zero because the driving force
' tential (Ere,)of a synaptic current is more positive than the on the ion, or ions, that can pass through the channels will
threshold of the postsynaptic cell, that synapse is excitatory be zero. In this case, when the synaptic channels open, Vm
(Figures 6-20A and 6-21A). If Ere"is more negative than will not change. In some cases, Erevis more positive than
threshold, the synapse is inhibitory. At fast chemical V,,, but more negative than threshold (Figure 6-21B). In
synapses, excitatory currents are typically carried through this situation, the postsynaptic potential is depolarizing, but
channels that conduct Na+ or Ca2+.These channels may be it is, nonetheless, inhibitory because it increases the diffi-
permeable to K+ as well, as is the ACh channel of the ver- culty of bringing Vmup to threshold. In each of these two
tebrate neuromuscular junction, but the K+ current special cases, the synapses have an inhibitory action, be-
itself does not contribute to the excitatory nature of the cause activation of these channels can counteract a simul-
synapse (see Figure 6-19). Inhibitory synaptic currents are taneous activation of excitatory channels (Figure 6-21C).
typically carried by channels that are permeable either to In effect, the opening of inhibitory postsynaptic channels
K+ or to C1-. The reversal potential, E,, ,for K+ or C 1 typ- "short-circuits" excitatory currents, because the positive
ically lies near V,, so it is more negative than the thresh- charge carried into the cell by excitatory currents can leave
186 P H Y S I O L O G I C A L PROCESSES
...............................................................................
' '1 Figure 6-20 Synaptic currents can be excitatory or in-
hibitory (A) Transmltter D evokes an excitatory depolarir-
---------- - - ---- - -- - -- - - - ing postsynaptic potential, because it increases an ionic
conductance generating a net inward current that adds
positive charge t o the cell's interior For example, trans-
mitter D might increase the permeability t o Na+. Trans-
C
a, mitter H produces an inhibitory hyperpolarizing synaptic
0 potential, because ~tIncreases the conductance t o ions
a
that cause a net loss of positive charge from the cell. For
example, transmitter H might increase the permeability to
- ----- ------ - - - - - --- ---- K+ or t o CI-. (B) The direction in which positive current
flows through channels opened by transmitter D is oppo-
site t o the direction in which ~ o s i t i v ecurrent flows
through channels opened by transm~tterH
0
In
Transmitter D Transm~tterH
the cell through the inhibitory channels, preventing the pos- more negative than V,, . When ACh is applied to H cells,
itive charges from bringing Vm up to threshold. it opens C1- channels, allowing C 1 to flow into the cell
Note that there is nothing inherently excitatory or in- down its electrochemical gradient. The result is to shift Vm
hibitory about any particular transmitter substance. Rathel; toward Ec,, hyperpolarizing the cell (see Figure 6-22A).
the properties of the channels that are opened by the trans- If all extracellular C 1 is replaced by SO:-, which cannot
mitter and the identities of the ions that flow through those pass through the chloride channels, application of ACh
channels, determine how a transmitter affects the postsyn- leads to an efflux of C1-, because it now has an outwardly
aptic cell. For example, ACh is an excitatory transmitter at directed electrochemical gradient. This efflux of negative
the vertebrate neuromuscular junction, where it opens charge produces both a depolarization and an increase in
channels that allow Na+ and K+ to cross the postsynaptic the frequency of action potentials (see Figure 6-22B). Thus,
membrane. In contrast, ACh is inhibitory at the terminals ACh is normally inhibitory for these cells, but it can pro-
of parasympathetic neurons innervating the vertebrate duce excitation if the electrochemical gradient for C1- is re-
heart, where it affects Kc-selective channels. versed. In fact, in this species of snail, there are other brain
From this description, it follows that an inhibitory cells (called D cells, or depolarizing cells) that naturally
transmitter could be made excitatory if the ionic gradients maintain a high intracellular C1- concentration by actively
across the postsynaptic membrane were changed. This ex- accumulating Clk. Acetylcholine causes an increase in gc,
perimental manipulation has been accomplished for neu- for these cells, as it does in the H cells. However, in the D
rons of the mammalian spinal cord and for neurons of a cells, the net effect is a depolarization, because the electro-
snail (Figure 6-22). In certain snail neurons, ACh increases chemical gradient for C 1 is normally outward. Hence, in
g,, of the postsynaptic membrane. In one group of these this example, excitation and inhibition depend critically on
cells (called H cells, or hyperpolarizing cells), the intra- the nature of ionic gradients, and not on the identity of the
cellular C 1 concentration is relatively low, making E,, signaling molecule.
n
COMMUNICATION ALONG A N D BETWEEN NEURONS 187
..............................................
A B &
+
C--
Action potential
Threshold
hE
- - - - - - E r etI,,,, ------- ~nhib
bE Threshold
Inhibitory
Rest~ngpotential
-
Excitatory
- - - - - - - - - - - Ewv, excit
Threshold Figure 6-21 Excitatory and inhibitory synaptic signals interact in the
postsynapticcell. (A) An action potent~alarises out of an excitatory post-
synaptic potential if that postsynaptic potential bringsthe membrane po-
tential V,, above threshold. (B) A postsynaptic potential is inhibitory,
even if it depolarizes V,, if its Ere,is more negative than the threshold for
Excitatory impulse generation.(C) An inhibitory transmitter (as in part B) may reduce
and the depolarizationproduced by an excitatory transmitter (as in part A) suf-
inhibitory ficiently to keep the postsynaptic potential from reaching threshold.
Presynaptic inhibition Experiments performed in the transmitter released from the terminal. In other examples of
1960s on neurons of the mammalian spinal cord and on the presynaptic inhibition, the inhibitory transmitter modifies
crustacean neuromuscular junction revealed an additional some property of Ca2+channels in the presynaptic mem-
inhibitory mechanism at some synapses. In this mechanism, brane, rendering them less responsive to de~olarization.Be-
called presynaptic inhibition, an inhibitory transmitter is re- cause the release of transmitter molecules depends on Ca2+
leased from a terminal that ends o n the presynaptic termi- entry into the terminal (seethe next section of this chapter),
nal of an excitatory axon (Figure 6-23). In this case, the reducing Ca2+entry reduces transmitter release. Regardless
presynaptic terminal of the excitatory axon is itself a post- of the mechanism, the net effect of presynaptic inhibition is
synaptic element. During presynaptic inhibition, the that the postsynaptic cell receives less transmitter and thus
amount of transmitter released from the excitatory terminal a smaller postsynaptic potential is generated.
is reduced, which reduces synaptic excitation in the cell that Postsynaptic and presynaptic inhibition produce quite
is postsynaptic to the excitatory neuron (see Figure 6-23B). different consequences for the postsynaptic cell. Postsyn-
In some cases, the presynaptic inhibitory transmitter in- aptic inhibition globally reduces the excitability of the post-
creases g, or g,, in the presynaptic terminals of the excita- synaptic cell, making it is less able to respond to all excita-
tory axon, which reduces the amplitude of any AP invading tory inputs. In contrast, presynaptic inhibition acts only on
the excitatory terminal and hence diminishes the amount of specific inputs to the cell, allowing the cell to remain
f ACh ( l o k 5 M)
SPOTLIGHT 6-4 then V , must b e
From this equation, if g, is greater than gNa,
, than t o EN,, and vice versa. Solving equation 4 for
closer t o E
CALCULATION OF V,,, = Ere"gives
REVERSAL POTENTIAL
The value of the reversal potential of an ionic current elicited by
From equation 5, it is apparent that Ere,will not b e simply the al-
a stimulus or a neurotransmitter depends on the relative con-
gebraic sum of EN, and E, but will lie somewhere between the
ductances of the ions carrying the current as well as on their
two, depending on the ratio gN,lgK.Thus, if g
,, and g, become
equilibrium potentials. If we assume that only Na+ and K+ carry
equal t o each other (e.g., as they may when endplate channels
current in response t o the stimulus, the reversal potential can
are activated by ACh in frog muscle), the membrane potential
b e related t o the conductances of these ions by using equation
will shift toward a reversal potential that lies exactly halfway be-
5-10, with the values g, and gNa
representing the respective tran-
tween g,=, and g,:
sient changes in the two conductances.
INa = 9
a, X (Vm- ENa) (2) For frog muscle, E, is about -100 mV, and EN,is about +60 mV.
Hence, we would predict that during synaptic activation of a frog
At the reversal potentlal, I, and INa
must b e equal and oppos~te
muscle, Ere, = (-100 + 60) = -20 mV The measured rever-
regardless of the relat~veconductances, because the net current
sal potential of the current at the frog neuromuscular synapse,
must b e zero. Thus, when V , IS at the reversal potent~al,Ere,,
- 10 mV, is somewhat more positive than this value, possibly be-
cause g
, is actually somewhat greater than g,.
-IK= lNa (3)
To summarize, the reversal potentials of membrane currents
Substltut~ngfrom equat~ons1 and 2, atthe reversal potentlal we differ according t o the species of ions that participate, the equi-
have librium potentials of those ions, and the relative conductances
t o each of the ionic species that participates in the current.
-gKWm - EK) = - ENa)
gNa(Vm (4)
normally responsive to other inputs. Thus presynaptic inhi- presynaptic impulse in producing a postsynaptic potential
bition provides a mechanism for narrowly targeted and change) among the many synaptic connections onto a par-
subtle control of synaptic efficacy (the effectiveness of a ticular neuron.
Inhibitory
terminal
Muscle
-t20 rns
Figure 6-23 Neurons that produce inhibition at the crustacean neuro- tential (epsp).(2) Stimulation of the inhibitoryaxon(labeled I on record)
muscularjunction also inhibit excitatory motor neurons presynaptically. produced a depolarizing inhibitory postsynapticpotential (ipsp)of about
(A) The morphological arrangement of excitatory and inhibitorytermi- 0.2 mL! (3) If the inhibitory neuron was stimulated afew millisecondsafter
nals, showing the location of an inhib~torysynapse that produces presyn- the excitatory neuron, the excitatory postsynaptic potential was unaf-
aptic inhibition and the arrangement for the experiment illustrated in fected. (4) However, if the ~nhibitoryneuron was stimulated a few mil-
part B. (B) lntracellular recordingfrom the muscle fiber innervated by ex- liseconds before the excitatory neuron, the excitatory postsynaptic po-
citatory and inhibitory motor neurons. (1) Stimulation of the excitatory tential was almost abolished. [Adapted from Dudel and Kuffler, 1961.1
axon (labeled E on record) produced a 2 mV excitatory postsynaptic po-
PRESYNAPTIC RELEASE OF tracellular Ca2+,or both, caused the evoked endplate po-
NEUROTRANSMllTERS tential to become smaller in amplitude. Katz and his co-
workers used this observation to find concentrations of
The properties presynap- these at which the evoked endplate potential be-
tic terminals determine the effectiveness of synaptic trans-
came as small as, or a small and simple multiple of, a single
mission, because the number of transmitter molecules spontaneously occurring miniature endplate potential.
affects the size the postsynaptic They then recorded postsynaptic responses to presynaptic
Understanding transmitter release is thus of central impor- motor nerve impulses in this high M $ + - ~ Cai+~ ~solution
tance for understanding synaptic transmission and its nor- and obtained the following statistical evidence:
ma1 role in neuronal communication. Besides its impor-
tance for physiology, the history of experimentation on Some motor nerve impulses produced n o response at
transmitter release provides classic examples of the scien- all. They called these trials "failures."
tific method and experimental strategies. A particularly
striking example is the demonstration, by Sir Bernard Katz Some impulses produced endplate potentials that had
and his co-workers, that neurotransmitters are generally re- approximately the same amplitude as those of single
leased in tiny packets called quanta. More recent experi- spontaneous miniature endplate potentials.
ments have shown that synaptic release is closely related to Some impulses produced endplate potentials with am-
other forms of exocytosis used by cells, such as glandular plitudes that were integral multiples (e.g., two, three,
cells, to release chemicals (see Chapter 9). The conservation four, etc.) of the mean amplitude of single spontaneous
of this mechanism has facilitated experiments designed to miniature endplate potentials (Figure 6-25).
understand the details of exocytosis in all cells.
These findings lent further support to the hypothesis that
Quantal Release of Neurotransmitters a normal endplate potential is produced by a large number
In their investigation of neuromuscular transmission, Paul of transmitter units (equivalent to the units producing the
Fatt and Bernard Katz (1952)discovered that spontaneous spontaneous miniature endplate potentials) released in uni-
"miniature" depolarizations (<I mV in amplitude) could son during a presynaptic action potential. Calculations
be recorded from the vicinity of the postsynaptic membrane showed that in frog muscle approximately 100-300 such
of the motor endplate in the frog muscle (Figure 6-24). units could account for the amplitude of normal evoked ex-
These spontaneous signals became progressively smaller citatory postsynaptic potentials.
when the intracellular recording electrode was inserted at Because synaptic release appeared to occur in the form
greater distances from the endplate, and they eventually of discrete units-packets, or quanta, of transmitter mole-
disappeared if the electrode was moved far enough away cules-Katz and his associates called the process quanta1
from the endplate. Because these potentials have a shape, release. They then asked another simple, direct, and im-
time course, and drug sensitivity similar to those of end- portant question: What was the makeup of the unit, or
plate potentials, they were called miniature endplate po- quantum, of transmitter release? Was it a single molecule of
tentials (mepps).The measurement of miniature endplate ACh? If not, how many molecules were in a quantum? If
potentials has played a key role in analyzing and under- the cause of a spontaneous miniature endplate potential
standing the mechanisms of neurotransmitter release. was a single ACh molecule that leaked out of the pre-
A key insight of Katz and his collaborators was to ask synaptic terminal, then adding a very low concentration of
how these spontaneous miniature potentials might be re- ACh to the saline bathing the muscle should greatly in-
lated to the endplate potentials elicited by motor nerve crease the number of miniature endplate potentials. Begin-
stimulation. They asked whether miniature endplate po- ning- with very low concentrations of ACh and working
tentials might represent a "unit" of transmitter release. If their way up to higher concentrations, they never saw an
so, endplate potentials evoked by stimulating the motor increase in miniature endplate potentials, but they did ob-
nerve could simply be the result of many such units being serve a depolarization in the postsynaptic muscle fiber that
released in unison subsequent to presynaptic APs. It had smoothly increased in size with increased ACh concentra-
been found earlier that progressively increasing the con- tion. They concluded that the miniature endplate potentials
centration of extracellular Mg2+ or decreasing that of ex- were not produced in response to single ACh molecules.
- 1 s
1 1 mv
Figure 6-24 Spontaneous miniature endplate potentials can be recorded from the motor
endplate region of a skeletal muscle fiber. Note that the amplitudes of the miniature endplate
potentials are small and variable.
190 PHYSIOLOGICAL PROCESSES
........................................
naptic vesicles (see Figure 6-14) could be the basis both for
the miniature endplate potentials (each caused by the re-
\\
lease of a single vesicle) and for evoked endplate potentials
LOW-ca2+
solution (each caused when many vesicles are released simultane-
ously). Various lines of evidence support this view, includ-
ing electrical measurements of membrane capacitance at
the presynaptic terminal, which increases during exocyto-
sis because vesicles fuse with the plasma membrane and in-
-
crease its surface area.
It is impossible to study individual quanta, so the quan-
t
Stimulus
50 rns
tal release of transmitter at the frog motor endplate has
been studied extensively by statistical analysis, and we now
have a coherent description of events that take place during
transmitter release. At any given time, only a fraction of the
population of vesicles inside a nerve terminal are available
for immediate release. For any particular set of physiolog-
ical conditions (e.g., Ca2+and Mg2+concentrations, tem-
perature), there is some particular probability that any one
of the available vesicles will be released. (In fast transmis-
sion, the available vesicles appear to be those located in the
active zones.) If the concentration of Ca2+ in the extra-
cellular fluid is reduced, entry of Ca2+into the presynaptic
terminal in response to an AP is reduced. An influx of Ca2+
into the terminal is essential for transmitter release; there-
fore, when the influx of Ca2+drops, so does the probabil-
ity that any presynaptic vesicle will be released. If the prob-
ability is sufficiently low, it produces the condition
illustrated in Figure 6-25B, in which presynaptic stimula-
tion leads to many failures (i.e., no vesicles are released) or
0 1 2 3 to the release of only one, two, or a few vesicles to produce
Units of amplitude endplate potentials with amplitudes that correspond to
Figure 6-25 Under appropriate conditions, action potentials in a motor multiples of one, two, three, and so forth, miniature end-
nerve produce small endplate potentials, similar to miniature endplate plate potentials. When the normal number of quanta in an
potentials, in the postsynaptic muscle fiber. (A) A nerve-muscle prepa- endplate potential, called its quantal content, is reduced by
ration is placed in a low-Ca2+solution, which reduces the amount of
a low concentration of extracellular Ca2+,it is possible to
transmitterthat is released at the neuromuscularjunctionwhen the nerve
is stimulated. The evoked endplate potentials then have small and vari- determine the number of vesicles that were released in re-
able amplitudes. (B) Amplitude distribution of spontaneous miniature sponse to each stimulus in a large series. Statistical analysis
endplate potentials (top) and of endplate potentials (bottom) that were of those numbers has shown that the probability of vesicle
evoked by motor newe stimulation in low-Ca2+saline. Note the many fail- release follows a Poisson distribution (see Figure 6-25B),
ures in the lower distribution. Most of the evoked endplate potentials had
which implies that release is a random process.
an amplitude distribution similar to that ofthe single spontaneous minia-
ture endplate potentials. The continuous curves in the upper and lower
histogramswere calculated from the theoretical Po~ssondistribution, as-
Depolarization-Release Coupling
suming that the evoked endplate potentials are made up of units corre- According to the quantal theory of transmitter release, the
sponding to the spontaneous miniature endplate potentials. [Part B probability that a given vesicle will undergo exocytosis and
adapted from Del Castillo and Katz, 1954.1
release its contents at any given instant is quite low when
the presynaptic membrane potential, Vm, is at the resting
level. When the presynaptic neuron is at rest, miniature
Indeed, they calculated that each miniature endplate po- endplate potentials are relatively rare and occur randomly;
tential was produced by the release of a packet of trans- however, when the presynaptic membrane is depolarized,
mitter molecules consisting of about 10,000 molecules of the probability of quantal release dramatically increases.
ACh, and that these molecules activated about 2000 post- The increased probability of release can be seen in the in-
synaptic channels. At about this time, electron-microscopic creased frequency of miniature endplate potentials that ac-
studies revealed the presence of membrane-bounded pack- companies a steady depolarization in the presynaptic neu-
ets, or vesicles, in presynaptic endings (see Figure 6-13C). ron (Figure 6-26).
The vesicles constituted the anatomical basis for the pack- The relation between presynaptic membrane potential
ets of transmitter that had been inferred physiologically by and transmitter release was examined by Bernard Katz and
Katz and his group. The release of transmitter from presy- Ricardo Miledi in an unusually large synapse onto the
C O M M U N I C A T I O N A L O N G A N D BETWEEN N E U R O N S 191
...............................................................................
When the presynaptic membrane was depolarized,
transmitter was released (detected as depolarization in
the postsynaptic cell) even though the normal mecha-
nism of the AP had been eliminated.
As the presynaptic membrane was made increasingly
depolarized, the amplitude of the postsynaptic poten-
tial increased, implying that the amount of transmitter
Figure 6-26 When the presynaptic terminal is depolarized, miniature released varied directly with depolarization of the
endplate potentials in the postsynaptic muscle fiber become more fre- presynaptic terminal: more depolarization caused more
quent. Depolarization of the presynaptic terminal by application of cur- transmitter to be released.
rent (lower trace) Increasesthe probability of transmitter release, shown
by an increase in the frequency of miniature endplate potentials recorded A given amount of presynaptic depolarization pro-
in the muscle fiber (upper trace). [Adapted from Katz and Miledi, 1967.1 duced a smaller postsynaptic response when the con-
centration of Ca2+ was reduced in the extracellular
fluid.
squid giant axon (Figure 6-27A). Because of the presynap-
tic terminal's large size, microelectrodes for passing current These three results supported the hypotheses that
and for recording membrane potential can be inserted into events in the presynaptic terminals depend on a depolar-
the terminal close to the synaptic region. This arrangement ization of the membrane, but not on the specific ions that
would be technically impossible in most other synapses, be- cause the depolarization, and that Ca2+ions play a role in
cause most presynaptic terminals are tiny. In this experi- neurotransmitter release. Further support for the hypoth-
ment, Na+ channels were blocked by tetrodotoxin (TTX), esis that Ca2+ions were responsible for transmitter release
and K + channels were blocked by tetraethylammonium came from an experiment in which the presynaptic termi-
(TEA), so V, of the presynaptic terminal could be con- nal was depolarized all the way to the Ca2+equilibrium po-
trolled without interference from all-or-none APs. The tential, ECa(seeNernst Equation in Chapter 5).In this con-
postsynaptic potential, recorded with a third microelec- dition, there could be no net flux through open Ca2+
trode inserted near the synapse, provided a highly sensitive channels in the presynaptic terminal, because there was no
bioassay of how much transmitter was released from the net driving force on Ca2+ions. Correspondingly, no trans-
presynaptic cell. The following results are shown in Figure mitter release was detected until the membrane potential,
6-27B and C: V,, was allowed to return to its resting level after the
Postsynaptic
neuron
Presynaptic
terminal
-I
I
I
I
I
I
Low current
I
I
I
I
n
High current
lrijected current
I
I
I
I
I
/ ~ a - a e q u o r i nluminescence
I I I I
Giant
axon
Postsynaptic voltage
* p -
Figure 6-28 The light-emitting protein aequorin showed that transmitter is released only when calcium ions
can enter the presynaptic terminal. In this experiment, Na+ and K+ currents at the squid giant synapse were
blocked with TTX and TEA. The Ca2+-indicatoraequorln was injected into the presynaptic terminal (light red),
and the terminal was stimulated with injected depolarizing current. Pre- and postsynaptic potentials were
recorded with intracellular microelectrodes.The responses to weak and strong presynapticstimulating currents
are shown at the right. A postsynaptic potential was recorded only when aequorin produced light, indicating
that CaZ+had entered the presynapt~cterm~nalSynapt~cves~cles ~nthe term~nalare shown as small red c~rcles
[Adapted from Ll~nasand N~cholson,1975 ]
Figure 6-29 Exocytos:~at the synaptlc rermi-
na can b e vls,a .zed w ~ t han elecrron m cro-
scope Exocytos s at a n e n e rermina of a frog
n e ~ r o m ~ s c ~jLnct
l a r on The arrows p o ~ ntto
veslc es emptying ,nto rne synapt c cleft.
[Courtesy of He-ser.1
through identifiable steps. Filled synaptic vesicles dock pref- 1. If the candidate substance is applied to the membrane
erentially at active zones after which a maturation process of a postsynaptic cell, it must elicit precisely the same
makes synaptic vesicles competent to undergo fast Ca2+-de- physiological effects in the postsynaptic cell as does
pendent membrane fusion. Exocytosis takes less than presynaptic stimulation.
0.3 ms, but only one of many docked vesicles at an active 2. The substance must be released during activity of the
zone fuses and not all APs lead to exocytosis from an active presynaptic neuron.
zone. Recently, Dieter Bruns and Reinhard Jahn (1995) 3. The action of the substance must be blocked by the
reported the first real-time measurement of transmitter same agents that block natural transmission at that
release from single synaptic vesicles in cultured leech neu- synapse.
rons that contain two kinds of vesicles. Small, clear vesicles
discharge about 4700 transmitter molecules with a time Using these criteria, physiologists have identified many
constant of about 260 ps, and large, dense-core vesicles other neurotransmitters, but enormous effort has been re-
release about 80 x lo3 molecules with a time constant quired. The identification of the transmitters at most
of about 1.3 ms. This technical advance suggests that synapses in the vertebrate central nervous system has been
molecular details of release may soon be discovered for very difficult, because very little transmitter is released at
many kinds of synapses, which will expand our under- most synapses (only about l o 4 molecules per synapse per
standing of how synaptic transmission regulates neuronal AP-compare this with Avogadro's number). Moreover,
communication. neuronal tissue is a nonhomogeneous collection of tightly
packed and diverse cell types, which complicates collecting
Nonspiking Release these molecules. However, a sufficient number of transmit-
Some neurons, in both vertebrates and invertebrates, have ter substances have been identhed that patterns have begun
been shown to release neurotransmitter from their termi- to emerge.
nals even in the absence of APs, which is called nonspiking The effect that a neurotransmitter exerts on a post-
release. At least some of these neurons are unusual in that synaptic cell can depend on either of two very different
they may never carry APs; all information transfer is ac- mechanisms, and this difference forms the basis of one clas-
complished by electrotonically conducted voltage signals. sification scheme for neurotransmitters. All transmitters ul-
The amount of transmitter that is released into the synap- timately modify the conductance of ion channels, but the
tic cleft by these cells depends on the membrane potential, change in conductance is produced in a variety of ways.
V,. When the cells are more strongly depolarized, they re- Some transmitters act directly on ion channel proteins to
lease more transmitter; when they are less strongly depo- change conductances through the postsynaptic membrane
larized, less transmitter is released. As a result, the amount and thereby to change V,; this type of transmission is fast,
of transmitter released is a direct function of the membrane or direct, synaptic transmission. Other transmitters work
potential, V,, of the neurons, which can indicate how through a biochemical pathway within the postsynaptic
strongly the neurons themselves were stimulated. cell, changing the state of membrane-associatedor cytoso-
lic second messengers and thus producing changes in ion
channel proteins. The shifts in V, generated by this second
THE CHEMICAL NATURE OF type of transmitter occur more slowly, so this type of trans-
NEUROTRANSMllTERS mission is slow, or indirect, synaptic transmtsston. We have
Once it became clear that most synaptic transmission is car- now identified more slow transmitters than fast ones. Neu-
ried by chemical signals, it became important to identify the rotransmitters that work indirectly may also act as neuro-
molecular identities of these transmitter substances. By the modulators, affecting many neighboring neurons, if they
mid-1960s, only three compounds had been unequivocally are released more generally into extracellular fluids and are
identified as neurotransmitters: acetylcholine, norepineph- thus able to modify the behavior of many postsynaptic neu-
rine, and y-aminobutyric acid (GABA).In the process of rons at once.
identifying and characterizingthese compounds, three cri- Alternatively, neurotransmitters can be sorted into
teria were established that had to be met by any candidate two groups based on their chemical structure. One group
molecule to determine that it was a neurotransmitter: consists of small molecules (Table 6-2). The other group,
194 PHYSIOLOGICAL PROCESSES
.........................................
TABLE 6-2 choline, glutamate, aspartate, and adenosine triphosphate
Small neurotransmitter and neuromodulatory molecules are (ATP)usually, but not always, participate in fast excitatory
distributed broadly across many phyla
synaptic transmission. y-Aminobutyric acid and glycine
Transmitter Site of action Action mediate fast inhibition. All of these transmitters, except as-
Acetylchol~ne Vertebrate NMJ E partate and ATP, have been shown to open ion channels in
(ACh) Vertebrate autonomic nervous the membrane of the postsynaptic cell, and it is expected
system pre- t o postgangl~onlc that aspartate and ATP will be found to do so as well.
neurons E Acetylcholine (Figure 6-30) is the most familiar of the
Parasympathet~cneurons Eorl established transmitter substances. Neurons that release
Vertebrate CNS E ACh, which are said to be cholinergic, are widely distrib-
Many ~nvertebrates E or l uted throughout the animal kingdom. To list just a few of
Norepinephrine Vertebrate postgangllonlc the known examples, ACh is the neurotransmitter used by
sympathet~cneurons E or l vertebrate motor neurons, the preganglionic neurons of the
Vertebrate CNS Eorl vertebrate autonomic nervous system, the postganglionic
Glutam~ca c ~ d Vertebrate CNS E neurons of the parasympathetic division of the autonomic
Crustacean CNS and PNS E nervous system, and many neurons of the vertebrate cen-
yAminobutyric Vertebrate CNS I tral nervous system. Acetylcholine is also the transmitter in
acid (GABA) a number of invertebrate neurons, including cells of the
Crustacean CNS and PNS I
Annelid CNS and PNS I
molluscan central nervous system, motor neurons of an-
Serotonin Vertebrate and invertebrate
nelid worms, and sensory neurons of arthropods. Mole-
(5-hydroxy- CNS cules that have crucial structural features in common with
tryptarnine) 1, M ACh can also act at cholinergic synapses (see Figure 6-30).
Dopamine Vertebrate, annelid, arthropod For example, many molecules, such as carbachol, can ac-
CNS, PNS, or both E or l (?) tivate cholinergic synapses; molecules that mimic the action
Glycine Vertebrate spinal cord I of a neurotransmitter are said to be agonists at the synapse.
Abbreviations: NMJ, neuromuscular junction; CNS, central nervous Alternatively, molecules that have structural features in
system; PNS, peripheral nervous system; E, excitatory; I, inhibitory; common with a transmitter can block transmission. For ex-
M, modulatory.
ample, D-tubocurarine, the active agent in the South Amer-
Phenylalanine
OH
neuron
Tyrosine
Circulation
NH, Figure 6-34 Epinephrine is synthesized from phenylalanine, with dopamine and norepi-
nephrine as intermediates,and is inactivatedby reuptake or by methylation. (A) Biosynthetic
pathway leading to epinephrine. Each of the lowerthree molecules is used as a neurotrans-
mitter by some neurons. (B) Norepinephrine is synthesized from the amino acid phenylala-
nine, through conversion into tyrosine, and stored in synapticvesicles. After it is released into
the synaptic cleft, some norepinephrine is taken back up Into the presynapticterminal, and
some is deact~vatedby methylat~onand carried away In the blood Cytoplasmic norepl-
OH nephrine is either repackaged Into synaptlc ves~clesor degraded by monoamine oxidase
3,4-Dihydroxyphenylalanine(dopa) (MAO) [Part A adapted from E~duson,1974, part B adapted from Mountcastle and
Baldessar~n~, 1968 ]
-
view showing the five subunits associated t o
2 nrn form the channel. These structural features
have been inferred on the basis of electron
microscopy and X-ray diffraction analysis.
[Adapted from Unwin, 1993.1
2 nrn
generated this sparse distribution by taking advantage of micropipette that had a smoothly polished tip with a tip di-
the changes that occur in frog skeletal muscle after the mo- ameter of 10 pm, which they filled with Ringer solution
tor nerve controlling the muscle has been cut. containing a low concentration of ACh or one of its ago-
When a muscle is denervated (i.e., it loses its neuronal nists. They then brought the pipette up to the surface of the
input-experimentally the axons are crushed), the region muscle fiber, exposing any AChRs under the pipette tip
of the membrane that responds to ACh gradually spreads to the ACh. The pipette was connected to a highly sensi-
across the surface. Initially, only the membrane at the end- tive, low-noise amplifier (Figure 6-36A) with which they
plate region can respond, but eventually most or all of the could record currents flowing in the extracellular pipette.
membrane contains AChRs and can respond to ACh. (The When it was applied snugly to the surface of the denervated
normal suppression of these extrajunctional AChRs is muscle fiber, the pipette detected minute (less than
thought to be dependent on two factors: first, trophic ac- 5 x 10-l2 A) and transient inward currents (Figure 6-36B)
tion from the motor neuron that innervates each muscle produced by the transient opening of the ACh-activated
fiber and, second, electrical and contractile activity that channels. With this experiment, Neher and Sakmann pro-
takes place in an innervated muscle fiber. If the motor axon duced the first recordings ever made of currents through
is allowed to reinnervate the muscle, the extrajunctional re- single ion channels in a biological membrane. Indeed, this
ceptors disappear, and sensitivity to ACh is again confined effort produced the first direct evidence that ionic currents
to the endplate.) cross the membrane through discrete, gated channels rather
The broad, but sparse, distribution of extrajunctional than by some other means, such as carrier molecules.
ACh-activated channels that develops in denervated frog Single-channelcurrents such as those first recorded by
muscle was exploited by Neher and Sakmann to explore Neher and Sakmann in 1976 are more or less rectangular
the gating of the channel, using their newly developed in shape; they turn on abruptly and then turn off abruptly,
patch-clamp method. The muscle membrane was voltage- and they are all-or-none.This observation strongly suggests
clamped (see Spotlight 5-3) at a hyperpolarized potential to that the channels can exist only in one of two states, com-
increase the driving force for inward current. They used a pletely shut or completely open. Moreover, the unitary cur-
C O M MU N l C A T l O N A L O N G AND BETWEEN NEURONS 201
. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
r-c-
I
Pipette current
Voltage c l a m p
these unitary currents indicate that the channels can fluctu-
ate between several closed states and at least one open state.
Binding of an agonist molecule to the receptor sites of the
closed channel greatly increases the probability that the
channel will change to an open state and briefly allow ions
to flow through the channel. The channel remains open for
only about 1 ms and then closes, even though ACh is still
bound to the receptor sites. After a short time, the agonist
molecules leave the binding sites, and the channel remains
closed until more molecules of ACh bind (Figure 6-37).The
macroscopic currents and postsynaptic potentials recorded
at a synapse represent the sum of many such single-channel
events in the postsynaptic membrane.
Denervated muscle cell
Other ligand-gated channels
Since the ACh channel proteins were purified from electro-
plax organs, several types of ligand-gated channels have
been isolated from neurons and characterized, including
the glycine, GABA,, and neuronal ACh receptors, all of
which mediate rapid postsynaptic responses. These recep-
tors have a common pentameric protein structure, and each
is composed of two to four different kinds of subunits. As
in the muscle ACh channel, only one type of subunit binds
Figure 6-36 The patch recording technique reveals ionic currents the ligand. The remarkable homologies among these dif-
through single acetylcholine receptor channels. (A) The muscle mern-
ferent channel proteins have allowed the diversity of sub-
brane is held at a hyperpolarized potential (-120mV) by a voltage-clamp
circuit, greatly increasing the driving force on ions through acetylcholine
unit types and their distribution in nervous tissue to be
receptor (AChR) channels, while the surface ofthe muscle is explored with characterized at the molecular level. Somewhat surpris-
a patch pipette filled with Ringer solution containing 2 X lo-' M suberyl- ingly, for each type of receptor-ACh, glycine, and
choline (an ACh agonist). (B) When the pipette tip is sealed tightly against GABA,-a number of different subunits are found to be
the membrane, brief, minute, inward currents are recorded. In this exper-
assembled in different combinations to produce receptors
iment, the pipette records current flow through the ion channel of a sin-
gle AChR protein complex that opens transiently when agonist molecules
with slightly different properties. Moreovel; each type of re-
are bound t o receptor sites. [Adapted from Neher and Sakmann, 1976.1 ceptor is expressed in a unique and characteristic pattern
within the mammalian brain, indicating that expression of
receptor subtypes is regulated differently in different re-
gions of the nervous system. Recognizing the large number
rents recorded from each nicotinic ACh-activated channel of permutations that are possible, even within receptors
are about the same size as the currents recorded from all that respond to a single neurotransmitter, has helped us to
other nicotinic ACh-activated channels, provided that the understand how subtle the mechanisms that allow the
electrochemical driving force is kept constant. Ohm's law brain to achieve its highly differentiated functional states
indicates that this result must mean that all individual nico- can be. Furthermore, a comparison of the DNA sequences
tinic ACh channels have similar conductances. When two of the receptors for ACh, GABA, and glycine reveals them
or more channels in the patch being recorded open with to be closely related, suggesting that all ligand-gated ion
overlapping times, the individual, unitary single-channel channels may have a common ancestral origin.
currents sum linearly, producing a current two (or three, DNA sequence analysis has revealed that glutamate
etc.) times as large as the individual unitary current. These receptors belong to a separate family having only a slight
currents are not present unless the pipette contains ACh or resemblance to the nicotinic receptors. Currently, there is
an agonist, and the frequency of their occurrence depends intense interest in this receptor family, both because gluta-
on the concentration of the transmitter or agonist in mate is the most common excitatory neurotransmitter in
the pipette. From Ohm's law, the conductance of a sin- the mammalian central nervous system and because gluta-
gle open nicotinic AChR channel was calculated to be mate receptors participate in modifications of synaptic
about 2 x 10-l1 S, which is usually expressed as 20 pi- strength, which may underlie learning and memory. At pre-
cosiemens (20 X 10-l2 S; i.e., the channels have a resistance sent, three types of fast-acting glutamate receptors have
of 5 x lolo a). been identhed and are named for their sensitivity to specific
Since the pioneering patch-clamp experiments of Neher agonists. The agonists that typify the three receptor classes
and Sakmann, many ligand-gated postsynaptic ion chan- are kainate, quisqualate (a-amino-3-hydroxy-5-methyl-
nels have been studied intensively with this method of isoxazole-4-propionic acid), and NMDA (N-methyl-D-
recording single-channel currents. Statistical analyses of aspartate). These receptor types are considered further later
202 PHYSIOLOGICAL PROCESSES
...........................................................................
4---- +
Open Closed
in this chapter in the section concerning mechanisms of this system provides yet another mechanism for producing
synaptic modification (see Long-term potentiation). subtle control within the nervous system.
A well studied example of indirect neurotransmission
Receptors in Slow, Indirect Neurotransmission that regulates ion channels is found in heart atrial cells, the
A large family of receptors responds to the family of slow system Otto Loewi used more than 75 years ago in the first
neurotransmitters. Interestingly, these receptors have many demonstration that neurons can transfer information by
features in common with receptors that respond to light, to way of chemical signals. Acetylcholine acts on muscarinic
odor, to hormones, and to other extracellular messengers. receptors in the heart to hold K+-selectivechannels open,
Most such receptors act by activating members of a group prolonging a hyperpolarization. Several different kinds of
of proteins, known as G proteins, that are associated with experiments were needed to establish that this action of
the cell membrane and that bind guanosine triphosphate ACh depends on a G protein.
(GTP). A G protein consists of three subunits, called a, P, Some of the experimental results are described here.
and y. The G protein-transmembrane signaling pathway Acetylcholine has been found to act on heart atrial cells
was discovered and described by Alfred Gilman and Mar- only if GTP is inside the cells, and the muscarinic activation
....-
tin Rodbell, who studied its role in transduction of non- of Kt channels is known to be blocked by pertussis toxin,
steroid hormone signaling (for a more complete treatment
of G proteins, see Chapter 9); for this work, they received Neurotransmitter
the Nobel Prize in 1994. When GTP binds to a G-protein Extracellular / Ion channel
molecule, the protein is activated, and it catalyzes the hy-
drolysis of the bound GTP to GDP, which terminates its ac-
tivation (Figure 6-38). When a membrane receptor mole-
cule binds to its ligand, this cycle of binding and
hydrolyzing GTP is facilitated, because the receptor-ligand
'
complex catalyzes the release of GDP from the G protein,
making the binding site more rapidly available to a new \
GTP molecule. Three separate proteins contribute to G G-protein complex \Second messengers,
other proteins
protein-mediated synaptic transmission. The neurotrans- \
mitter receptor molecules span the membrane, binding the t
neurotransmitter on the extracellular face and catalyzing Other cell functions
G-protein activation on the cytoplasmic face. The activated
Figure 6-38 lntracellular second messengers modify channel conduc-
G protein can regulate the activity of effector proteins, tances at slow chemical synapses. G proteins participate in signal trans-
which can be ion channels or enzymes that control the con- mission at many slow chemical synapses. At this kind of synapse, the re-
centrations of intracellular second messengers or both. We ceptor protein spans the plasma membrane. Neurotransmittermolecules
now know of more than 100 receptors that act through G bind to the extracellulardomain of the receptor, which activates a G pro-
tein that resides on the cytoplasmic s~deof the membrane. The act~vated
proteins, and these signaling molecules respond to a wide
G protein regulates the activity of other intracellular proteins, which di-
variety of external stimuli ranging from peptides to light rectly or indirectly changes the conductance through ion channels in the
and odors. The G proteins themselves constitute a family of membrane.Activated G proteins can also modify other cellular functions,
at least 20 different proteins. The combinatorial richness of changing metabolic pathways or the structure of the cytoskeleton.
B
5 pM G,*
Control
r 5 0 pMGa*
,A C h
/
Patch of Extracellular .Muscarinic r e c e ~ t o r
atrial cell
membrane
GnRH
C cell B cell v ACh
50 mV
-
9th and 10th
Current chain gangha
Figure 6-41 Both muscarinic acetylcholine receptors and GnRH recep- t o the fast epsp. (C)Cholinergic neurons from the seventh and eighth
tors depolarize a postsynaptic cell by closing M-type potassium ion chan- spinal nerves Innervate C cells of the ninth and tenth sympathetic gan-
nels. (A)When acetylcholine (ACh) binds to muscarinic receptors or when glia, whereas neurons from the third, fourth, and fifth nerves innervate
the GnRH-like neuropeptide binds to its receptor, M-type channels close, only B cells in those ganglia. Only C cells receive terminals that are im-
reducing the K+ current across the membrane and depolarizing the neu- munoreactive for GnRH, but stimulating the seventh and eighth spinal
ron. (B) The effect of a fast excitatory postsynaptic potential (epsp) in a nerves produces a late, slow epsp in both B and C cells, suggesting that
postsynapticB cell before, during, and after a slow epsp. During the slow GnRH diffuses from its site of release at the surface of C cells and acti-
epsp, the decrease in the Kt current through M channels increased the vates receptorson B cells. [Part B adapted from James and Adams, 1987;
excitability ofthe B cell, producing a train of action potentials in response part C adapted from Jan and Jan, 1982.1
muscarinic receptors, repolarization of the membrane Within the past few years, studies in the crustacean
by the K+ current is prevented and further excitation is stomatogastric ganglion have demonstrated the extreme
potentiated. The late, slow excitatory postsynaptic poten- power of neuromodulatory mechanisms. This ganglion
tial acts similarly, but with a longer latency and for a longer contains only 30-40 identified neurons, whose intercon-
time, and it shares the M channel as a final common path- nections have been characterized in detail and whose out-
way. However, there is an additional twist because the put patterns are well known. When certain neuromodula-
peptide neurotransmitter diffuses to nearby neurons, which tory substances, such as proctolin or cholesytokinin, are
it can influence identically if the appropriate receptors are added to the saline bathing the stomatogastric ganglion, the
present (Figure 6-41C). Only some of the presynaptic neu- properties of at least some of the membrane channels
rons can release GnRH, but most postsynaptic cells seem change dramatically, effectively rewiring the entire ganglion
to have GnRH receptors, which strongly suggests that and generating circuits and outputs that are never seen in
neuromodulation is a normal part of this neuronal cir- the absence of the modulator. Thus, neuromodulators af-
cuitry. Taken together, these mechanisms can produce a va- ford a means of remodeling neuronal circuitry, allowing a
riety of postsynaptic effects following presynaptic trans- set of neurons to interact in several distinctly different
mitter release. A brief burst of activity in the presynaptic ways, even though their physical synaptic relations remain
cells would typically generate only the fast excitatory post- unchanged.
synaptic response. More prolonged stimulation might acti-
vate the slow pathway in addition, which would effectively
INTEGRATION AT SYNAPSES
amplify the response of the postsynaptic cell to its fast ex-
citatory postsynaptic potentials. With still greater stimula- Only rarely are single neurons responsible for producing
tion, the late, slow pathway would additionally increase behavior. Even the simplest behavior requires that several
the effectiveness of fast excitatory postsynaptic potentials hundred to many thousands of neurons act in a coordi-
and could also potentiate responses in neighboring neurons nated fashion. This coordination among neurons is called
(see Figure 6-41C), increasing the efficacy of neurotrans- neuronal integration. Used in this sense, "to integrate"
mission in cells that are not directly postsynaptic to the means "to combine into a whole." At the level of a single
GnRH-releasing neurons. Moreover, this modulation could neuron, integration consists of responding to incom-
be relatively long lived, given the long time constant of the ing synaptic inputs either by producing an AP or by not
late slow, response. producing one, and every neuron integrates the various
excitatory and inhibitory synaptic signals that impinge on How do the many thousands of individual synaptic in-
it. The process of integration depends heavily on the pas- puts onto a motor neuron influence its activity? Synaptic
sive electrical properties of the membrane that lies between currents spread electrotonically from synapses on dendrites
the synapses and the spike-initiating zone. In addition, the and the soma. How much the current decays over dis-
density and voltage sensitivity of the Na+ and K+ channels tance is determined by the cable properties of the neuron
determine the threshold and the rate of firing that is pro- (Figure 6-43), but in all cases, synaptic potentials become
duced in response to a given synaptic depolarization. smaller as they spread away from their sites of origin and
Much of what we know about neuronal integration has toward the spike-initiatingzone (see Passive Spread of Elec-
been obtained from studies of the large a-motor neurons trical Signals earlier in this chapter and Figure 6-16). Be-
(Figure 6-42) in the vertebrate spinal cord. These neurons cause the decrement depends on distance, a synaptic current
innervate groups of skeletal muscle fibers at neuromuscu- set up at the end of a long, slender dendrite will decay more
lar junctions. In vertebrates, these are the only neurons that than will currents closer to the spike-initiating zone, so dis-
synapse directly onto skeletal muscle fibers, so they play an tant synapses exert a relatively smaller influence on the ac-
exceedingly important role in generating overt behavior tivity of the postsynaptic neuron. As a result, the location of
(see Chapter 10).Thousands of inhibitory and excitatory synapses, as well as the initial size of synaptic currents, can
synaptic terminals contact the dendrites and cell body of influence how much control particular synapses have. (In-
each a-motor neuron. The net effect of all synaptic activity terestingly, recent evidence suggests that in at least some
is to control the frequency with which APs are generated in neurons of the mammalian brain, there may be some Na+
the cell. This frequency of firing (typically measured in im- channels in dendritic membranes, and these channels can
pulses per second) determines the strength of contraction in boost synaptic currents, preventing them from decaying as
the set of muscle fibers innervated by the motor neuron. rapidly as they would if they were conducted only electro-
All of the integrative activity in a neuron is centered on tonically.) In many cases, the density of inhibitory synapses
producing APs (i.e., excitation) or suppressing them (i.e., is highest near the axon hillock, where these synapses can
inhibition). Because APs are the only events that can carry be most effective in preventing excitatory synaptic current
information over distances greater than a few millimeters, from depolarizing the spike-initiating zone to threshold.
only synaptic inputs that cause APs in a-motor neurons can We have learned many of these concepts from experi-
generate behavior. Any excitatory input that fails to bring ments in frogs of the genus Rana. For example, in such an
a motor neuron to threshold, either by itself or by summa- experiment, several segments of the spinal cord of an anes-
tion with other inputs, is lost because no AP is produced in thetized frog are exposed by opening up the vertebral col-
the postsynaptic cell and the signal dies out. umn. Then a microelectrode is lowered into the ventral
In an a-motor neuron, APs are generated in the initial horn of the gray matter and inserted into the soma of a sin-
segment of the axon just beyond the axon hillock (see Fig- gle a-motor neuron. Small bundles of afferent axons dis-
ure 5-2). This region is more sensitive to depolarization sected from the dorsal root are placed on silver-wire, stim-
than are the soma and dendrites (perhaps the membrane ulating electrodes, providing stimulation to some axons
has a higher density of Na+ channels in this location), so it that cause the a-motor neurons to be excited and to oth-
has a lower threshold for producing APs. If it is to generate ers that cause the motor neurons to be inhibited.
APs in the cell, a synaptic current must be able to bring the Initially, the intracellular recording electrode will pick
membrane of the spike-initiating zone up to threshold. up randomly occurring postsynaptic potentials. These sig-
d
\I
Myelin
sheath
I Threshold
Action potential
--
-
.-m
C
(I) Synaptic potential
0
a
Distance
Figure 6-43 Each synaptic input decays with distance as it travels to the initiating zone of the axon hillock (or at the first node of Ranvier), where
spike-initiating zone. An excitatory postsynaptic potential originating in the firing threshold is lowest. The graph shows the relative values of the
a dendrite spreads electrotonically and gets smaller with distance (top). threshold potential and the synaptic potential along the membrane be-
The density of Na' channels (red dots) in the membrane determines the tween the synapse and the spike-initiating zone. The dashed line shows
threshold (blacktrace at bottom) for generating an AP The synaptic po- what the amplitude of the excitatory postsynaptic potential would be if
tential gets smaller as it spreads toward the axon, and no AP is generated the AP were blocked.
until the current reaches the dense Na+-channeldistribution in the spike-
nals are caused by synaptic input onto the motor neuron junction acts as a single relay synapse, transmitting in a one-
that is not under experimental control. Typically, the activ- to-one manner (i.e., one postsynaptic impulse for each
ity consists of synaptic potentials with amplitudes of about presynaptic impulse), a motor neuron requires the more-or-
1mV, which are similar to those of miniature endplate po- less concurrent activation of numerous excitatory synaptic
tentials recorded at the muscle endplate (see Figure 6-24).It inputs impinging on it in order for the synaptic potential to
has been shown that stimulation of a single neuron that is reach the firing threshold for the initiation of a postsynap-
presynaptic to these motor neurons releases only from one tic AP. Thus, the decision to fire is a response to a collection
to several quanta of transmitter in response to a presynaptic of presynaptic inputs, and although each small synaptic cur-
AP. In this respect, excitatory synapses ending on a motor rent is ineffective by itself, the activity at a single ending can
neuron are quantitatively different from those on a neuro- contribute significantly to the integrative behavior of the
muscular junction, where a single motorneuron terminal re- neuron. This rather democratic behavior prevents activa-
leases approximately 100 to 300 quanta in response to a tion of motor neurons by trivial input or spontaneous ac-
single presynaptic impulse and produces an excitatory post- tivity in their input neurons. More importantly, it provides
synaptic potential of 60 mV or more. The transmitter re- a means of integrating inputs from various sources, both ex-
leased from a single synaptic ending onto an a-motor neu- citatory and inhibitory, to determine when the neuron will
ron depolarizes the neuron by only about 1 mV, far less produce APs and how many there will be.
than the amount required to shift the membrane potential As the strength of the stimulus current applied to the
to the firing level. Whereas the vertebrate neuromuscular presynaptic axons in the dorsal root is increased, more and
208 P H Y S I O L O G I C A L PROCESSES
........................................
more excitatory axons become active; that is, they are ye-
cruited by the increased stimulus. When these neurons fire in
unison, the total amount of transmitter released onto the
motor neurons rises, producing more individual synaptic
currents that add together to cause a larger excitatory post-
synaptic potential. When inputs from several individual
synapses add together simultaneously to change Vm in the synaptic Inhibitory
current -\-/I, synaptic
postsynaptic neuron, the process is called spatial summation. current
The summed synaptic inputs can lead to greater depolariza-
tion if they are all excitatory (Figure 6-44). If inhibitory
transmitter is released simultaneously with excitatory trans-
mitter, it also produces synaptic currents that sum with the
excitatory currents (Figure 6-45). Open inhibitory synaptic
channels can short-circuit the depolarizing current carried by
Na+ ions moving through excitatory channels; that is, as de-
polarizing positive charge is carried into the cell by Na+ ions,
some of that charge is promptly removed from the cell when
K+ ions move out or CI- ions move in through inhibitory
synaptic channels. The activation of inhibitory .
synapses
. - re-
duces the depolarization at the spike-initiating zone and de-
creases the probability that an AP will be produced. '1 [ - - - - - - - a and b
When a second postsynaptic potential is elicited within
a very short time after the first, it can add to, or "ride pig- Figure 6-45 Excitatory and inhibitory synapticcurrents sum. Stimulation
of separate presynaptic pathways gives rise to excitatory (a) and inhibitory
gyback" on the first, even if the two synaptic events are
(b) synaptic currents. Traces at lower right show synaptic potentials
caused by the same presynaptic neuron. This effect is called recorded from the spike-initiating zone when either a o r b was stimulated
individually and then when they were stimulated together, illustratingthe
effects of summation. The dashed arrows indicate that some of the exci-
tatory synaptic current is diminished by the open inhibitory channels.
role are (1) changes in the presynaptic terminals and Figure 6-48 Synaptic facilitation occurs at the frog neuromuscularjunc-
(2) changes in the postsynaptic neuron. One example of a tion. In this experiment, curare in the bathing saline blocked some ACh
presynaptic mechanism would be a change in the amount receptors, reducing the amplitude of exc~tatorypostsynaptic potenti&
to below the firing threshold. Two st~muliwere delivered to the nerve in
of transmitter released from presynaptic terminals in re-
rapid succession. The second synaptic potential summed with the falling
sponse to a presynaptic AP. An example of a postsynaptic phase of the first, producing a larger postsynaptic potential; but, in ad-
mechanism would be a change in the postsynaptic appara- dition, the amplitude of the second response (indicated by the line la-
tus that altered the amplitude of depolarization produced beled 2) was greater than could be accounted for by summation alone.
COMMUNICATION ALONG A N D BETWEEN NEURONS 211
...............................................................................
A
tained by Katz and Miledi (1968).They used a carefully
positioned micropipette to supply pulses of Ca2+ ions
to the external solution near a motor endplate of a frog
muscle that was immersed in Ca2+-free Ringer solution
(Figure 6-49A). They found that facilitation of the post-
synaptic potential evoked by the second stimulus was great-
est when a pulse of extracellular Ca2+ions was supplied to
coincide with the arrival of the first AP (Figure 6-49B). The
first pulse of Ca2+did not significantly enhance facilitation
if it was given after the first AP arrived at the terminals
B S t ~ m u l u st o Postsynaptic (see Figure 6-49B).Thus, if synaptic facilitation is to occur,
I ?
-
Ca2+
Ca2+must be available to enter the presynaptic terminal
when an AP invades the terminal. If Ca2+ions can enter the
terminal from the external fluid, Ca2+ions from the second
AP add to any Ca2+ions remaining from the first, leading
to the release of more transmitter.
Homosynaptic Modulation:
Posttetanic Potentiation
When a frog motor axon is stimulated tetanically (i.e., at a
high frequency for a relatively long time), synaptic trans-
mission at the neuromuscular junction is initially depressed
after the stimulation. However, responses to test pulses ap-
plied at later times after the stimulation are found to be
larger than normal. This increase in the amplitude of the re-
sponse lasts for as long as several minutes and, during this
Figure 6-49 Synaptic facilitation depends on the presence of calcium
time, the responses are said to be potentiated. This post-
ions in the extracellular fluid. (A)The motor neuron innervating the mus-
cle fiber was stimulated, and the resulting postsynaptic potential was
tetanic potentiation is another example of a use-dependent
recorded. The bathing solution was calcium free, but small pulses of change in presynaptic efficacy, and it is found in one
CaCI, were provided by a CaC12-containingpipette positioned just at the form or another at many types of synapses. Figure 6-50 il-
endplate region. In this experiment, the relative timing between stimuli lustrates the results of one such experiment. Initially,
to the motor neuron and the delivery of CaCI, was vaned. ( 0 )Recordings
of postsynaptic potentials in the muscle fiber. Horizontal black bars show
the timing of Ca2+pulses. Thin vertical lines indicate stimuli to the pre-
synaptic neuron.Trace 1 shows the amplitude of a postsynapticpotential
in response to a single AP in the motor neuron. In the three other traces,
-
the temporal relation between the first AP and the pulse of CaCI, was var-
ied. In all cases, Ca2+ions were available at the time of the second AP. Fa-
cilitation occurred only when Ca2+ions were present at the endplate
when both of the APs reached the endplate. [Adapted from Katz and
H
Miledi, 1968.1
50.s-' 5 min
$: 200
The best evidence indicates that synaptic facilitation de- +-
5
.-
pends on the amount of free Ca2+within the presynaptic .I
terminal. The concentration of intracellular free Ca2+ions r +g 100
0
n
rises in the terminal when the first AP opens voltage-
dependent Ca2+channels, and this increase in Ca2+ion 0 H
concentration persists for a short time. When the second 50 s-'
impulse arrives at the te5mina1, the Ca2+ concentration
Figure 6-50 Tetanic stimulation of a frog motor nerve elicits depression
is still somewhat elevated,and the ca2+ ions that enter as a
and potentiation of excitatory postsynaptic potentials in muscle fibers.
of the second add the remaining Ca2+ions, Curare was used to reduce the amplitude of synaptic potentials, block-
generating an even higher Ca2+ concentration in the ing the produaion of APSand revealing the amplitude of the synaptic PO-
terminal. Because the release of transmitter is a Dower func- tentials. When the nerve and muscle were bathed in normal frog Ringer's
tion of the intracellular Ca2+ concentration near the pre- solution, which has a ca2+concentration of about 2 rnM (top), stirnulat-
ing the motor nerve at 50stimuli per second for about a minute produced
synaptic release sites, this small increase in Ca2+concen-
first a depression in subsequent excitatory postsynaptic potentials and
tration inside the terminal produces a large increase in the then When the concentration of extracellularCa2+was re-
amount of transmitter released subsequent to the second duced to 0 225 mM, only potent~at~on was seen after h~gh-frequency
impulse. Experimental evidence for this hypothesis was ob- st~mulatlon.[Adapted from Rosenthal. 1969 I
212 P H Y S I O L O G I C A L PROCESSES
......................................
excitatory postsynaptic potentials (epsps)were evoked at a but not at, a presynaptic ending, they are said to act het-
frog neuromuscular junction by stimulating the motor nerve erosynaptically, because transmission through the synapse
at a low control rate (one stimulus every 30 seconds). The is altered by an additional, third neuron, which released
rate of stimulation was then increased to 50 per second for the modulator. One class of heterosynaptic action that
a period of 20 seconds, after which a series of test stimuli has already been discussed in this chapter is presynaptic in-
were administered at the initial rate of one every 30 seconds. hibition; another, in which the amount of transmitter re-
In Ringer solution that contained a normal concentration of leased is increased by the presence of the modulator, is
Ca2+ (see Figure 6-50, top), posttetanic depression of the called heterosynaptic facilitation.
evoked epsps occurred immediately after the tetanic stimu- In heterosynaptic modulation, the modulator is
lation. However, within 1minute, the amplitude of the ep- thought to alter the number of Ca2+ions that enter the ter-
sps increased; in other words, posttetanic potentiation had minals subsequent to a presynaptic AP. Synaptic modula-
occurred. The amplitude of the epsps returned to the con- tors usually do not directly open (or close) ion channels. In-
trol level after about 10 minutes. In Ringer solution that stead, they change how ion channels respond to another
contained a lower-than-normal concentration of Ca2+(see stimulus; by doing so, they increase or decrease ionic cur-
Figure 6-50, bottom), there was no depression, and the rents carried through channels that are activated by a pre-
posttetanic potentiation subsided more rapidly. synaptic AP. This action by modulators is typically medi-
These results are thought to depend on events within ated by one or more intracellular messengers that act on the
the terminals. During high-frequency stimulation in a nor- ion channels. In contrast, fast neurotransmitters bind to
mal concentration of extracellular Ca2+ (1.8 mM), the membrane receptors and open (or close) channels.
available synaptic vesicles are released faster than they can The most extensively studied example of heterosynap-
be replaced, so the amount of transmitter available for re- tic modulation at a synapse is found in the sea hareAplysia
lease is depleted and remains low for a time immediately af- californica, a sluglike gastropod mollusk that has been
ter the high-frequency stimulation. Later in the posttetanic widely used in studies of neuronal plasticity. Eric Kandel
period, quanta of transmitter available for release are re- and his associates have found that excitatory transmission
stored, and the depression subsides. During tetanic stim- between specific identified neurons in the central nervous
ulation, Ca2+ ions that have entered the terminals ac- system of Aplysia is enhanced during behavioral sensitiza-
cumulate, load up the available Ca2+-binding sites that tion. They have found that this enhancement occurs
ordinarily buffer the intracellular concentration of Ca2+, through heterosynaptic facilitation of transmitter release
and linger within the terminals until they are gradually triggered by the release of serotonin near the synapse
pumped out by active transport across the cell membrane. (Figure 6-51). In this case, serotonin is thought to elevate
It is believed that posttetanic potentiation and its slow de- the concentration of the intracellular messenger 3',5'-cyclic
cay reflect this increase and subsequent decrease in the con- adenosine monophosphate (CAMP),which has been found
centration of Ca2+inside the terminals. In low-Ca2+Ringer to influence the opening of a specific type of K+ channel,
solution, fewer Ca2+ions are available to enter the termi- known as the S channel. Specifically, when CAMP isele-
nals, so fewer synaptic vesicles can bind to the membrane vated in the presynaptic neuron, S channels are more likely
and release transmitter. As a result, there is less depletion of to be shut at any given Vm. The efflux of K + through
available synaptic vesicles, and there is no posttetanic de- S channels contributes to repolarization after an AP, so
pression. Posttetanic potentiation is just as pronounced, be- the closing of S channels will prolong the presynaptic AP
cause repeated stimulation does bring Ca2+ions into the and allow more Ca2+ions to enter the terminal through
terminals, but the potentiation decays more rapidly, per- voltage-gated Ca2+channels. An increase in the influx of
haps because the concentration of Ca2+inside the terminals Ca2+ ions allows more transmitter to be released and
is less elevated or because the presynaptic terminal is able increases the amplitude and duration of the postsynaptic
to pump the extra Ca2+out more rapidly because less has potential.
accumulated.
Long-Term Potentiation
Heterosynaptic Modulation In the past few years, intense interest has been focused on
The release of transmitter from nerve terminals can be in- long-term changes in synaptic efficacy that have been iden-
fluenced at some synapses by the presence of certain neu- tified in the mammalian hippocampus, the site of certain
romodulators. These modulatory agents include serotonin memories. High-frequency stimulation of inputs to the hip-
in mollusks and in vertebrates, octopamine in insects, and pocampus produces an increase in the amplitude of exci-
norepznephrine and GABA in vertebrates. All of these tatory postsynaptic potentials recorded in the postsynaptic
agents are also neurotransmitters (see Table 6-2). In addi- hippocampal neurons. In an intact animal, the increased
tion, endogenous opioids have been shown to act as mod- amplitude can last for hours-even days or weeks-after
ulatory agents in vertebrate neurons. Such agents, released the potentiating stimulation. This prolonged facilitation of
into the circulation or liberated by nerve endings near a synaptic transmission, called long-term potentiation, has
synapse, are believed to modify the release of transmitter been shown to occur in many synaptic pathways. At dif-
from presynaptic terminals. When they are liberated near, ferent sites, long-term potentiation may require different
C O M M U N I C A T I O N A L O N G A N D BETWEEN NEURONS 213
....................................................
Facilatatory
neuron
B
Modulatory-
transmitter
[CAMP]
increases
- S-type
K+ channels
- AP is
prolonged
- Ica
Increases
+ More
transmitter
blnds to close in synapse at terminal is released
receptor I I
In sensory neuron
After
stimulation of
facilatatory
Control After c A M P
injection
3 8
Number of
- '-
open
channels O - ~ ~ l ~ i I~ l I\
'' '-
j''- 3%
al< mu?%+-----d PA
2
Figure 6-51 Heterosynaptic facilitation at a synapse in Aplysia occurs events at the sensory neuron terminals. (C) An AP produced in the sen-
when potassium ion channels are closed in the synaptic terminals, allow- sory neuron (upper records) produces an excitatory postsynaptic poten-
ing more calcium ions to enter and causing more transmitter to be re- tial in the motor neuron (lower records). Stimulating the facilitatory neu-
leased. (A) If the facilitatory neuron is active at the same time that APs ar- ron prolongs the AP in the sensory neuron and produces a concomitant
. rive at the terminals of the sensory neuron, the net result is an increase facilitation of the synaptic response in the motor neuron. (D) Currents
in the amount of transmitter released by the sensory neuron. The trans- through single channels in the presynaptic membrane of sensory neu-
mitter of the facil~tatoryneuron binds to a receptor that causes an in- rons, recorded by the patch-clamp technique. Records were made be-
k crease in the level of CAMP in the terminal. Elevated CAMP causes S-type fore and after cAMP was injected into the neurons. The activity of S-type
K+ channels in the terminal to close, prolonging the depolarization of the K+ channels was reduced after cAMP was injected. [Part C adapted from
AP and holding voltage-gated Ca2+ channels open. (B) Summary of Kandel et al., 1983; part D adapted from Siegelbaum et al., 1982.1
patterns of stimulation, may decay at different rates, and tors is required for the induction of long-term potentiation,
may depend on different underlying mechanisms. In the although it is not required for normal neurotransmission.
cases studied, glutamate or a similar substance, has been To date, only a handful of experiments have been per-
found to be the principal excitatory transmitter. Of the formed in intact animals, but the results have been consis-
three pharmacologically distinct glutamate receptors (see tent with the hypothesis that changes in the properties of
Other ligand-gated channels earlier in this chapter), only NMDA receptors must occur if the strength of the synaptic
the type that responds to N-methyl-D-aspartate (NMDA) connection is to be modified. Furthermore, the results of
is thought to take part in long-term potentiation. At many several experiments suggest that, in long-term potentiation,
synapses in the hippocampus, activation of NMDA recep- a retrograde signal from the postsynaptic cell travels back
214 PHYSIOLOGICAL PROCESSES
...............................................................................
Glutamatergic Glutamatergic
neuron neuron
Figure 6-52 Long-term potentiation in the hippocampus depends on ulated tetanically by another source, the Mg2+blockof the NMDA chan-
NMDA receptors, a class of glutamate receptors, in the postsynaptic nels is removed, allowing Nat, Kt, and Ca2+ions to flow through the
membrane. The proposed role of NMDA receptors in long-term poten- NMDA channel. (C)The resulting increase in cytoplasmic Ca2+concen-
tiation at a hippocampal dendritic spine. Glutamate is released from the tration within the dendr~t~c spine activates intracellular second messen-
presynapticterminal and binds to both NMDA and quisqualate/kainate gers, including kinases, which can produce long-term modification of the
(QIK) type glutamate receptors in the postsynaptic membrane. (A) Ifthe quisqualate/kainate channels, and nitric oxide synthetase, which cat-
postsynaptic dendrite is not depolarized, Na+ and K+ flow through the alyzes the increased production of nitric oxide (NO). Nitric oxide is .
Q/K receptor channel, but not through the NMDA receptor channel, be- thought to diffuse back to the presynaptic terminal and potentiate the re-
cause Mg2+ions block NMDA channels when V, is near Vre, (B) When lease of transmitter in response to subsequent APs.
the dendrite is depolarized, as it is when the postsynaptic neuron is stim-
to the presynaptic terminal, producing a change in the Propagation along an axon depends on two phenom-
behavior of the presynaptic cell. Recent experiments sug- ena: (1)the longitudinal spread of current, which depends
gest that one such retrograde messenger could be the gas ni- on the cable properties of the axon; and (2) the continual
tric oxide (NO, Figure 6-52).Nitric oxide may act in the regeneration of the signal by excitation of new Na+ chan-
presynaptic terminal by modifying the activation of en- nels when resting membrane is depolarized by local-circuit
zymes, such as guanylyl cyclase or ADP ribosyltransferase. current as it flows along the axon ahead of the impulse. Be-
Whether this mechanism can account for the modifications cause APs are propagated without decrement along axons,
in neuronal function that underlie learning remains to be they can carry information between even widely separated
seen, but recent progress in identifying the molecular sub- parts of the nervous system. The velocity at which APs are
strates of memory has been encouraging, and the topic con- propagated depends on axon diameter and on the presence
tinues to be the target of an enormous research effort. (in some vertebrate axons) of insulating segments of
myelin sheath that are separated by nodes of Ranvier-
short, bare sections of axonal membrane. In myelinated
axons, saltatory conduction passes from node to node,
Within a single neuron, information can be carried in two skipping the intimately sheathed parts of the axon that lie
ways: (1)by graded, passively conducted potential changes between the nodes and thereby increasing the velocity of
and (2)by all-or-none, regenerative action potentials (APs). conduction.
Between neurons, information is typically carried by chem- There are two major types of synapses: electrical and
ical messenger molecules, although there are some electri- chemical. The principle of electrical synaptic transmission
cal synapses as well. Within a neuron, graded potentials oc- is essentially identical with that of impulse propagation; the
cur at specialized regions of the cell-for example, sensory current flows from one cell into another through low-resis-
receptor and postsynaptic membranes. Action potentials tance channels in specialized regions called gap junctions,
are confined largely to axons and axon terminals. Infor- depolarizing the second cell. There are two kinds of chem-
mation about signal intensity is coded by the amplitude of ical transmission: fast and slow. In fast synaptic transmis-
graded potentials and by the frequency of APs. sion, presynaptic endings liberate a neurotransmitter that
interacts with ligand-gated ion channels on the postsynap- Some changes in synaptic efficacy are due to prior activ-
tic membrane, causing the channels to open and allowing ity in the same synapse, which is called homosynaptic mod-
ionic current to flow, which produces a synaptic potential ulation. In some cases, it has been shown that a change in
across the postsynaptic membrane. In slow synaptic trans- synaptic efficacy occurs when the amount of transmitter that
mission and in a variant called neuromodulation, neuro- is released in response to a presynaptic AP changes. Modu-
transmitters bind to receptors, changing the state of G latory agents released from a third neuron or from endocrine
proteins associated with the membrane and indirectly mod- glands also can alter the effectiveness of synaptic transmis-
ulating the function of ion channels. Ion channels can be di- sion by influencing the amount of Ca2+entering the termi-
rectly regulated by G-protein subunits. Alternatively, nal during the presynaptic impulse and thereby increasing
neurotransmitters can modify the function of ion channels the amount of transmitter that is released from the terminal.
indirectly through
- cyclic nucleotides or through - cyclic
. nu-
cleotide-activated protein kinases that phosphorylate in-
REVIEW QUESTIONS
tracellular domains of the channels.
Chemical synapses have three advantages over electri- Compare and contrast the two basic kinds of signal
cal transmission: (1)the postsynaptic current can produce transmission found in the nervous system.
either an excitatory or an inhibitory action; (2)the post- Action potentials are carried along neurons by elec-
synaptic membrane is the source of the synaptic current, trical currents. Why are they so much slower than
and hence a tiny presynaptic axon can produce large post- electricity traveling along a wire?
synaptic currents through the influence of its transmitter on H o w can an AP travel over long distances without
postsynaptic channels; and (3)there is a greater possibility decrement, whereas synaptic potentials cannot?
of synaptic integration. Explain why, if all else is equal, an axon of large di-
In excitatory synapses, the transmitter changes the ameter will conduct impulses at higher velocity than
membrane's ion permeability, tending to make V, more will an axon of small diameter.
positive than the threshold for the initiation of an AP. In- Calculate the relative conduction velocities for un-
hibitory neurotransmitters change the conductance so as to myelinated axons that are 10 p m and 25 p m in di-
prevent V, from reaching threshold. The properties of ex- ameter, with all other parameters being equal in the
citation and inhibition are not inherent in the transmitter two kinds of axons.
substances; rather, excitation and inhibition depend on the Explain why, if all else is equal, a myelinated axon
ion selectivity of postsynaptic channels activated by the will conduct impulses at a higher velocity than will an
transmitters and on the reversal potentials of the currents unmyelinated axon.
carried through those channels. Fast neurotransmitters can Explain how loss of myelination, which happens in
act within milliseconds, and their effect is transient. Slow the demyelinating disease multiple sclerosis, disrupts
neurotransmission or neuromodulation changes V,, of the signal transmission in the nervous system.
postsynaptic cell for seconds or minutes, rather than for Design an experiment to test whether a synapse be-
milliseconds. tween two neurons is electrical or chemical.
Both excitatory and inhibitory transmitters are stored What determines if a neurotransmitter is excitatory
in, and released from, vesicles in the nerve terminal. The or inhibitory?
arrival of an AP depolarizes the presynaptic membrane, al- What factors determine whether a transmitter
lowing Ca2+ to enter the terminal. The Ca2+ ions, in a depolarizes or hyperpolarizes the postsynaptic
manner that is not yet completely understood, increase the membrane?
probability that synaptic vesicles will fuse with the termi- Marine invertebrates typically have much higher con-
nal membrane and release their contents into the synaptic centrations of inorganic ions in their body fluids than
cleft. Vesicular membranes subsequently undergo endocy- do freshwater invertebrates. For example, the table
a tosis and are recycled as new vesicles. Vesicles containing below gives the intracellular and extracellular K+
neurotransmitters that are responsible for fast neurotrans- concentrations for two molluscs: Limnaea, a fresh-
mission release their contents into the narrow synaptic water snail, and Sepia, the marine cuttlefish.
cleft between pre- and postsynaptic neurons; neuromod- Limnaea Sepia
ulatory transmitters are typically released from the side of lntracellular 14.8 mM 188.3 mM
the synaptic terminals.
Extracellular 1.8 m M 21.9 m M
Temporal and spatial summation of synaptic potentials
depends on the passive electrical properties of the post- If these two species had in common a neurotrans-
synaptic cell, and the net effect of all synaptic currents is to mitter that opened K+ channels in the postsynaptic
determine whether the membrane at the spike-initiating membrane, how would the values for Ere"at those
zone will be sufficiently depolarized to reach threshold. The synapses compare in the two species? If the post-
time constant of the postsynaptic cell allows temporal sum- synaptic neurons rested at - 70 mV and had a thresh-
mation to occur, even when the synaptic currents d o not old voltage of -55 mV, would the transmitter be ex-
overlap in time. citatory or inhibitory in each of the two species?
216 P H Y S I O L O G I C A L PROCESSES
.........................................
How can a synapse produce a depolarizing post- Hille, B. 1992. Ionic Channels in Excitable Membranes. 2d
synaptic potential and still be inhibitory? ed. Sunderland, Mass.: Sinauer. (An authoritative sur-
What is the evidence that an endplate potential is vey of the original work that revealed the mechanisms
composed of smaller units called miniature endplate of axonal conduction.)
potentials? Hille, B. 1994. Modulation of ion-channel function by
What places an absolute limit on the amplitude of a G-protein-coupled receptors. Trends Neurosci.
postsynaptic potential? 17531-536.
What prevents ACh released from the presynaptic Hodgkin, A. L. 1964. The Conduction of the Nervous Zm-
terminal from persisting and interfering with subse- pulse. Springfield, Ill.: Thomas. (A summary of the orig-
quent synaptic transmission? What happens if ACh inal work on how APs are initiated and conducted,
remains in the synaptic cleft? written by one of the primary contributors to the field.)
The amplitude of postsynaptic potentials decays with Kaczmarek, L. D., and I. B. Levitan. 1987. Neuromodula-
distance, so where on a neuron is the most effective tion: The Biochemical Control of Neuronal Excitabil-
site for a synapse to be located? ity. New York: Oxford University Press. (A summary of
Compare and contrast fast and slow chemical this field written by two experts.)
neurotransmission. Levitan, I. B., and L. K. Kaczmarek. 1997. The Neuron: Cell
What is meant by neuromodulation? and Molecular Biology. 2d ed. New York: Oxford Uni-
Discuss the role of Ca2+ in each of the follow- versity Press. (This has particularly good chapters on
ing events: depolarization-release coupling, facilita- synaptic transmission and neuromodulation,written by
tion, post-tetanic potentiation, heterosynaptic researchers who have contributed greatly to this field.)
modulation of transmitter release, long-term Nicholls, J. A., R. Martin, and B. G. Wallace. 1992. From
potentiation. Neuron to Brain. 3d ed. Sunderland, Mass.: Sinauer.
How is signal intensity encoded in graded signals, (A comprehensive treatment of function in the nervous
such as receptor potentials and synaptic potentials? system.)
How is intensity encoded in APs? Snyder, S. H. 1985. The molecular basis of communication
Compare the synaptic effects of ACh at the neuro- between cells. Scientific American 253:114-123.
muscular junction and on heart atrial cells. (An introduction to the process of chemical synaptic
transmission.)
Sudhof, T. C. 1995. The synaptic vesicle cycle: A cascade
SUGGESTED READINGS
of protein-protein interactions. Nature 375:645-653.
Cooper J. R., F. E. Bloom, and R. H. Roth. 1996. The Bio- (A recent review of vesicular neurotransmitter release.)
chemical Basis of Neuropharmacology. 7th ed. New Unwin, N. 1989. The structure of ion channels in mem-
York: Oxford University Press. (A classic book branes of excitable cells. Neuron 3:665-676. (An
describing the chemistry of neurotransmission and overview of the structure of ion channels in neurons
neuromodulation.) and muscles.)
.
Hall, Z. H. 1992. Molecular Neurobiology. Sunderland, Unwin, N. 1995. Acetylcholine receptor channel imaged in
Mass.: Sinauer. (A thorough description of the molecu- the open state. Nature 373:37-43. (High resolution
lar basis of signal transmission along axons and across images of the nicotinic acetylcholine receptor channel
synapses.) as it opens.)
L
!
L
!
s SENSING T H E E N V I R O N M E N T
Detection Mechanismsthat select stimulus modality: filters, Mechanismsthat select stimulus modality: filters,
1 carriers, tuning, inactivation carriers, tuning, inactivation
~rn~ilflcat~on Positlve feedback among chemlcal reactions or Posltlvefeedback among cells
I
membrane channels
Slgnal/noise enhancement
Slgnal/noise enhancement
1
Encoding and discrimination
Active processes in membranes
Intensity coding Different dynamic ranges among cells
Temporal differentiation Independent coding of quality and intensity
Quality coding Center-surroundantagonisms
1
Adaptat~onand termlnatlon Desenslt~zatlon
Opponent mechanisms
Temporal d~scrim~nat~on
I
Gating of ion channels
J.
Electrical response of membrane
Negative feedback
Temporal dlscrlmlnatlon
Repetitive responses
Channels open or close
Depolarization or hyperpolarization
J.
Transmission to brain .Electrotonic spread Spatial patterns: maps and image formation
Action potentials; number and frequency Temporal patterns; directional selectivity, etc.
Synaptic transmlsslon
- -- " - "- - -" -.-- -
*Arrows lndlcate that these operations are a serles of steps
fundamental differences in the roles played in the life of an By far, the most detailed knowledge available is about
animal by different sensory modalities. the molecules responsible for detecting photons: the protein
Recent evidence indicates that in vertebrates the recep- opsin and its associated molecules (Figure 7-3), which will
tors for three of the senses-vision, olfaction, and, proba- be discussed in more detail later in this chapter. However,
bly, sweet and bitter tastes-have in their cell membranes the close relation among sensory receptors was recently un-
receptor proteins with a common structural motif. The sec- derscored when the DNA sequence that codes for opsin
ondary structure of these membrane proteins includes was used to identify putative olfactory receptor molecules
seven transmembrane a helix domains, and transduction (Chess et al., 1992).The sequences in this new family of ol-
in all three senses requires G proteins as ~ntermediaries factory receptor molecules are expressed only in the cells of
(Figure 7-2). This pattern is also found in several neuro- the olfactory epithelium, and the family appears to be very
transmitter receptor molecules, including the muscarinic large (containing several hundred different gene prod-
acetylcholine receptor (see Chapter 6). ucts-perhaps even as many as a thousand). The sequences
Light
Neurotransmitter
/ \
G protein Effector Second-messenger protein Effector Second-
enzyme molecule enzyme messenger
Figure 7-2 The molecular mechanism of sensory reception in visual re- interacts with a GTP-binding protein (G protein)to alter intracellular sec-
ceptors resembles the molecular mechanism of transmission at many ond-messenger pathways. The second-messengersmodify conductance
synapses. For the examples shown, both processes begin with a struc- through ion channels, eitherdiredly or indirectly, and can thus modify the
tural change in a transmembrane protein (the receptor molecule),which pattern of APs in afferent neurons. [Adaptedfrom Bear et a1.,1996.]
SENSING THE ENVIRONMENT 221
...............................................................................
i
Figure 7-3 The visual pigment rhodopsin consists of a protein, opsin, cated inside the transmembrane domain of the opsin protein, near the
and an associated light-capturing molecule, 1I-cis-retinal. This d~agram center of the bilayer. Light is captured by 1I-cis-retinal, which causes the
[ shows rhodopsin as it would appear from outside the cell. The seven-
helix motif of this transmembrane receptor is found in other sensory re-
molecule to change into the all-transconfiguration, initiating a cascade
of intracellular events that finally produce a change in conductance
I
ceptor proteins and in receptor molecules that respond to hormones or through an ion channel (see Figure 7-53A).
6 to neurotransmitters (see Figure 7-2). The II-cis-retinal molecule is lo-
of these molecules differ significantly from one another ductance through membrane ion channels. When channel
only in a particular region that is thought to be the site at conductance changes, it can shift the probability that the
which stimulus molecules bind. neuron will produce an AP, although it must be remem-
Detection of salt and sour tastes occurs by way of much bered that not all receptors transmit information through
simpler mechanisms than those underlying vision and ol- the use of APs. In photoreceptors, cGMP may act directly
faction. Detection of these tastes depends on ion channels on a class of membrane channels to increase their conduc-
that are found on cells throughout the body. Sour sensation tance. The corresponding mechanisms for olfaction and
is mediated by a common pH-sensitive K+ channel, and the taste (also called gustation) are still unknown, although
salt response is caused by passive movement of Na+ across channels that respond to cyclic nucleotides have been re-
the cell membrane, depolarizing the taste cell directly. In cently found in the olfactory system.
both of these cases, because the stimuli are themselves ions Responses within a single receptor neuron encode in-
and are in great abundance, no intermediate amplification formation about the intensity of a stimulus, but they can-
step is needed. not directly report the quality of the stimulus. For example,
In some sensory systems, amplification of sensory sig- a single photoreceptor cannot report whether a stimulating
nals occurs within the receptor cells, mediated by a number light is red or blue. Additional information, such as the
of different intracellular mechanisms that will be discussed wavelength of light or the frequency of a sound, is con-
in more detail later in this chapter. Interestingly, this ampli- veyed by activity patterns within combinations of receptor
fication often occurs at the same time that noise is sup- cells that are activated by the stimulus. Typically, sensory
pressed, so the signal-to-noise ratio improves in the process. organs contain a variety of receptor cells that respond dif-
Amplification is best understood for vertebrate photore- ferentially to stimuli with different qualities. For example,
ceptors (largely from the eyes of cattle). When a photon is certain photoreceptors respond maximally to red light,
captured by a visual pigment molecule (discussed later in whereas others respond maximally to blue light. Thus,
this chapter), the net effect is to activate transducin, which when receptor cells are grouped into organs, significantly
is a GTP-binding protein, or G protein (see Figure 7-2 and more information about the stimulus can be conveyed, in-
Chapter 9). Transducin, in turn, activates a phosphodi- cluding its absolute intensity, its spatial distribution, and
esterase to hydrolyze cyclic guanosine monophosphate other characteristics such as quality.
a
(cGMP), which in turn leads to modification of conduc- Each sensory system must be able to detect stimuli that
tance through ion channels. Each photon captured leads to persist in time, while at the same time retaining the ability
the hydrolysis of many cGMP molecules, producing a huge to respond to further changes. The process of adaptation,
amplification in the signal. Although these steps have not described in Chapter 5 for single neurons, also occurs in the
yet been positively identified in the detection of odor and response of many receptor cells. From the perspective of an
taste, several facets of the transduction cascade are proba- organism, adaptation allows detection of new sensory stim-
bly similar. In each case, the energy available from a unitary uli in the presence of ongoing stimulation, and it thus
stimulus at the receptor site is so low that amplification makes the sensory system much more useful. For example,
within the receptor cell is required to generate neuronal wearing clothing stimulates touch receptors at all points
impulses that can carry the signal into the central nervous where our garments touch the skin, and we typically adapt
system. to the touch input from our garments. Yet, we can easily de-
Encoding sensory information into a neuronal signal to tect any new touch stimuli that impinge on our skin, even
be transmitted to the brain depends on changes in the con- at locations covered by our clothing. Many mechanisms
222 PHYSIOLOGICAL PROCESSES
...................................................
Figure 7-4 Stretch receptors in the tail of a
crayfish transmit information about the
Stimuli amount that the tail muscles are stretched.
The receptor consists of a sensory neuron
%1
Muscle
q ) v ~ e n d r i t e s
that has its stretch-sensitive dendrites em-
bedded in a special muscle bundle located
on the dorsal surface of the tail muscles.
When the tail of the crayfish bends, the mus-
Integration cle is stretched, and the receptor is activated.
Receptor potentials On the left side of the diagram, intracellular
records from the soma and extracellular
recordsfrom the axon show how the receptor
Weak Strong
Spikeinitating, 11 Conversion from
graded signal
into APs
responds to small and large stretches of the
muscle. The parts of the neuron are function-
ally d~fferentiated,as labeled at the right.
Graded receptor potentials from the stretch-
sensitive membrane of the dendrites are con-
Impulse
conduction verted into all-or-none APs at the spike-initi-
ating zone. The arrow indicates the direction
Impulse frequency
of AP propagation.
underlying adaptation take place within individual recep- Moreover, the size of the receptor potential varies with the
tor cells, and several of them appear to depend on Ca2+ strength of the stimulus, in contrast with APs that are all-
(e.g., in vision, olfaction, and mechanoreception). In addi- or-none. In these respects, receptor potentials resemble
tion, some adaptation depends on negative feedback from excitatory postsynaptic potentials at the postsynaptic
higher brain centers. , membrane of muscle and nerve cells and are quite distinct
from APs.
From Transduction t o Neuronal Output
Electrical measurements are important sources of insight
into the steps that lie between sensory transduction and the
generation of neuronal responses. One of the first such ex-
periments was done on receptor cells, called stretch recep-
tors, that sense muscle length in the abdomen of crayfish
and lobsters (Figure 7-4). Because each stretch receptor is
a relatively large cell, its soma can be impaled with micro-
electrodes. It is also possible to record extracellularly from
the axon of the cell. The dendrites of each stretch receptor
are attached across the surface of muscle fibers, and if the
muscle is stretched, a steady train of impulses can be
recorded from the axon. The frequency of the APs varies di- Stimulating
rectly with the amount of stretch applied. To understand current
the source of the APs, the intracellular potential can be
recorded by inserting a microelectrode into the cell body. B
A small stretch applied to the relaxed muscle leads to In normal saline In l T X
a small depolarization called the receptor potential (see
Figure 7-4). A stronger stretch produces a larger depolar-
Stretch stimulus 7 1
izing receptor potential. This change in Vmindicates that an lntracellular
underlying receptor current must flow across the mem- record
brane and that this receptor current must carry positive Extracellular T
record
charge into the cell to produce the depolarization. If recep- Jlo mv
tor potentials are sufficientlylarge, they trigger one or more 2s
APs in the cell (Figure 7-5).
Figure 7-5 The response of some crayfish stretch receptors IS phaslc, the
What is the relation between stimulus, receptor cur- response of others IS tonlc (A) Responses of a phasrcstretch receptor to
rent, receptor potential, and APs? Action potentials can be a weak (left) and to a strong (rrght) st~mulusA stronger stlmulus gener-
eliminated (seeFigure 7-5B) by blocking the electrically ex- ates more APs than a weak stlmulus Even when the stlmulus IS maln-
cited sodium channels with tetrodotoxin. When APs are tamed, the cell produces only one or a few APs (6) Responses of a tonlc
stretch receptor In normal sallne (left) and after the addltlon of tetro-
blocked, the receptor potential remains, indicating that
dotoxln (TTX, rrght) Tetrodotoxln blocks APs, revealing the underlying
it must be produced by a different mechanism from the receptor potent~al[Part A adaptedfrom Eyzagu~rreand Kuffler, 1955, part
one that generates the all-or-none upstroke of the AP. B adapted from Loewensteln, 1971 ]
Sensory receptors differ in how faithfully they repro-
duce the timing of a stimulus. A phasic receptor produces
APs during only part of the stimulation-often only at the
onset or at the offset of the stimulus-and thus cannot by
itself convey information about the duration of the stimu-
lus. In contrast, tonic receptors continue to fire APs Sp~ke-~n~t~ating
zone
throughout the stimulation and can thus directly convey in-
APs recorded
formation about the duration of the stimulus. (See Recep- by extracellular
-
tor Ada~tationlater in this chapter for a more complete electrode
discussion.) Local stimulation of the stretch receptor cell
was used to test the ability of various parts of a single cell
stimulus c;iY
to produce sustained trains of APs. In these experiments Stimulus current L
(Figure 7-6), a steady stimulus current produced a sus-
tained discharge only when the current depolarized the
low-threshold, spike-initiating zone of the receptor. When
other regions of the cell were stimulated, APs were pro-
duced, but there was no sustained train of impulses.
This difference implies that the ion channels at the spike-
timus 1 1
initiating zone have different properties from ion channels
kF
along the rest of the axon.
In summary, a general sequence of steps leading from
a stimulus to a train of impulses in a sensory neuron can Recording
be formulated from the results obtained with the crayfish electrode
stretch receptor (Figure 7-7).Stimulus energy produces an
alteration in a receptor protein, generally located in a mem- Figure 7-6 Sustained stimulation of a crayfish stretch receptor produces
brane. The receptor protein may be part of an ion channel, a long train of APs only when the stimulus depolarizes the spike-initiating
or it may modulate the activity of membrane channels in- zone. Other areas of the cell adapt rapidly to steady stimulation.
[Adapted from Nakajima and Onodera, 1969.1
directly through an enzyme cascade, which amplifies the
signal. In either case, the absorption of stimulus energy by
a receptor molecule eventually causes a population of ion creases, more channels respond, producing an increased (or
channels to open or to close. This change in membrane per- decreased) receptor current, and hence a larger receptor
meability produces a shift in V,,, in accord with principles potential. Thus, all the steps leading to, and including, the
presented in Chapter 5. As the intensity of the stimulus in- receptor potential are graded in amplitude. Unlike the
Stimulus reaches receptor cells Figure 7-7 Several steps l~nkthe onset of a
Transduction
1
Receptor protein is activated
st~mulusto the production of APs In a sensory
pathway In some sensory systems, the receptor
cells themselves generate and carry APs Into the
1
- Cascade of protein interactions
central nervous system (lower left) In other sys-
tems, receptor cells synaptically modulate APs
In other neurons that carry the s~gnalInto the
modifies intracellular second- central nervous system (lower rrght)
messengers
Amplification
1
- Ion channels open (or close)
I Graded
J. events
Change in conductance produces
a receptor current
1
Receptor current changes V,,
n
Change in V, spreads electrotonically Change in amount of transmitter
to spike-initiating zone released from receptor cell
Action Figure 7-8 Sensory signals are carried into the central nervous sys-
potent~al tem by action potentials. In two types of primary sensory neurons (A
and B), the receptor current arises in the receptor membrane and
Receptor spreads electrotonically to depolarize the spike-initiating zone. In
potentlal both cell types, the axon of the receptor cell extends into the central
nervous system. The difference between cells A and B is that the cell
body of cell A is located relatively far away from the central nervous
system, whereas that of cell B is located near the central nervous sys-
Stimulus tem. Many invertebrate sensory neurons are like cell A. Vertebrate
touch receptors are like cell B. (C) In this arrangement, the receptor
cell does not produce impulses but instead releases transmitter at a
synapse that modulates AP produalon by an afferent neuron. This
arrangement is found in the mammalian auditory and visyal systems.
Red arrows show the pattern of current flow, and the records at the
upper right were made by an intracellular electrode in the soma of
\
b
Spike-initiating zone neuron A.
A Ampl~tudelimited
Frequency limited
I by refractqry period
U 0
_= L
E la
a X
?! L -
Log of s t ~ m u l u s~ n t e n s ~ t y Amplitude of receptor potential Log of stimulus intensity
Figure 7-9 The response in most sensory receptors is proportional to the ceptor neuron depends linearly on the amplitude of the receptor poten-
logarithm of stimulus intensity (A) In many receptors, the amplitude of tial. The refractory period of the neuron sets an upper limit on the fre-
the receptor potential is linearly related to the logarithm of stimulus in- quency. (C) As a result of the relationsillustrated in parts A and B, the fre-
tensity over a large-but finite-range. The amplitude of the receptor quency of APs in many sensory axons varies linearly with the log of the
potential cannot increase infinitely, because it is limited by the reversal stimulus intensity The broken parts of the curves in parts B and C indicate
potential of the receptor current and by other biophysical properties of that refractoriness limits the maximum frequency of APs.
the receptor cell. (B) Within a certain range, the frequency of APs in a re-
226 P H Y S I O L O G I C A L PROCESSES
.........................................
and the amplitude of the receptor potential is explained, at scene are far more informative to a viewer than is the ab-
least in part, by the Goldman equation (see Chapter S), solute energy content of each stimulus. Thus, a deer in a
which predicts that Vm should vary with the log of the field or in a forest is acutely attuned to movements (changes
membrane's permeability to the ion (or ions), Pi,, involved in the distribution of visual stimuli) independent of changes
in the receptor potential. After stimulation, the change in in the lighting conditions.
Vm should be proportional to the log of the change in the
permeability to sodium, (P,,), that was produced by the Range Fractionation
stimulus. Normal stimulus intensities lie within the loga- The dynamic range of a multineuronal sensory system is
rithmic part of the input-output curve (see Figure 7-9A). typically much broader than is the range of any single re-
Not all receptors follow this general rule. In some recep- ceptor or afferent sensory fiber. The extended dynamic
tors, there is, instead, a power-function relation: the log range of the entire system is possible because individual af-
of the response amplitude is proportional to the log of ferent fibers of a sensory system cover different parts of the
the stimulus intensity. For practical purposes, either a log- full spectrum of sensitivity. The most sensitive receptors
arithmic function or a power function describes the rela- produce a maximal response at stimulus intensities that are
tion between stimulus intensity and receptor response very below threshold or only slightly above threshold for other,
well in the range of stimulus intensities commonly encoun- less sensitive receptors in the population. Above that inten-
tered by animals. The differences between the two func- sity, the most sensitive receptors become saturated, but the
tions become apparent only at extreme values of stimulus less sensitive receptors can take over at the higher intensi-
intensity. ties. Thus, at the lowest stimulus energies, a few especially
As a consequence of the logarithmic relation between sensitive sensory fibers will respond weakly. If the stimu-
the intensity of a stimulus and the amplitude of the recep- lus energy is increased a little, their discharge frequencies
tor potential, any given percentage of change in stimulus in- will increase, whereas new, less sensitive fibers will join in
tensity evokes the same increment of change (i.e., same weakly. With still greater stimulus intensities, another, for-
number of millivolts) in the receptor potential over a large merly quiescent lower-sensitivity population of afferents
range of intensities. In other words, a doubling of the stim- will join in. As the stimulus intensity is increased, receptors
ulus intensity at the low end of the intensity range will that are less and less sensitive will become active, a phe-
evoke the same increment in the amplitude of the receptor nomenon called recruitment, until the least sensitive sen-
potential as will a doubling of the intensity of the stimulus sory fibers will finally be recruited, and all receptors will re-
near the high end of the range up to the limit where the re- spond maximally. At that point, the system will be
ceptor potential cannot increase. So we have saturated and therefore unable to detect further increases
in intensity. This range fractionation, in which individual
receptors or sensory afferents cover only a fraction of the
total dynamic range of the sensory system (Figure 7-10),
enables the sensory processing centers of the central ner-
where I is the stimulus intensity and K is a constant. The vous system to discriminate stimulus intensities over a
logarithmic relation between the intensity of the stimulus range much greater than that of any single sensory recep-
and the intensity of the response (Figure 7-9C) thus "com- tor. One example of range fractionation are the photo-
presses" the high-intensity end of the scale, which greatly receptors of the vertebrate eye. Rod photoreceptors are
extends the range of discrimination. This relation is similar more sensitive to light and respond to dimmer stimuli;
to that which governs subjectively perceived changes in cones respond to bright light that saturates the rods (see Vi-
stimulus intensity, a relation known in psychology as the sual receptor cells of vertebrates later in this chapter).
Weber-Fechner law.
This feature of sensory systems confers great advan- Control of Sensory Sensitivity
tage. For example, this property allows us to recognize the How accurate are our sensations? That is, how do sense
objects in a particular scene, even when we see the scene organs compare with physical transducers such as ther-
under very different lighting conditions. If we observe the mometers, light meters, and strain gauges? From our own
scene in very bright sunlight, each object is distinguished by experience, we know that biological sensory systems may
its relative brightness. If we observe the same scene by not be very trustworthy indicators of absolute energy
moonlight, the absolute brightness of each object is very levels. Moreover, many sensations change over time. For
different from its brightness in sunlight-in fact, the dif- example, when a person dives into an unheated pool for a
ference in an object's brightness under the two conditions swim, the water initially feels colder than it does a minute
of lighting may be far greater than the difference in bright- or two later. A pleasantly sunny day may seem painfully
ness of various objects in the scene when illuminated by bright for a few minutes after a person emerges from a
bright sunlight. However, we are able to recognize the ob- dimly lit house; for this reason, even an experienced pho-
jects in the scene on the basis of their relative intensities, in- tographer requires a light meter to make accurate judg-
dependent of the absolute level of illumination. Detecting ments of camera exposure settings. These changes of
relative intensities and changes in intensity within a given perceived intensity, when the intensity of the stimulus
Figure 7-10 Range fractionation extends the dynamic
range of sets of sensory receptors Each curve In thls
a b c graph represents the dlschargefrequency of an indlv~d-
ual sensory afferent, plotted as a funct~onof stlmulus In-
tensity In thls hypothetical example, each of the four
sensory fibers, labeled a through d, has a dynamlc range
of about three to four log unlts of stimulus ~ntenslty,
whereas the overall dynamlc range of the four neurons
taken together covers seven log unlts of ~ntenslty
has not itself changed, are lumped under the general term held in the new position. In contrast, phasic receptors adapt
sensory adaptation. Where does the adaptation take place? quickly. In one class of phasic receptors, for example, APs
There is no simple answer. Some adaptation takes place occur only during changes in the strength of the stimulus,
in the receptor cells, some as a result of time-dependent as illustrated in Figure 7-1 1B by a mechanoreceptor that
changes in accessory tissues, and some in the central fires only when the displacement of the hair changes, and
nervous system. the frequency of APs depends on the rate of change.
Adaptation can take place at any of a number of dif-
Mechanisms of adaptation ferent stages in the processes that link a stimulus to the out-
Different classes of receptors exhibit different degrees put of a sensory neuron (Figure 7-12):
of adaptation. Tonic receptors continue to fire steadily
in response to a constant stimulus, as illustrated in 1. The mechanical properties of the receptor cell may act
Figure 7-11A, which shows a receptor that responds to the as a filter that preferentially passes transient, rather
displacement of a hair; this receptor produces APs at an al- than sustained, stimuli. This mechanism is common
most constant frequency when the hair is displaced and among mechanoreceptors.
A Displacement detector 6 Velocity detector Figure 7-11 A displacement detector fires toni-
cally, whereas a velocity detector fires phasically.
(A) Behavior of a tonic displacement detector.
This mechanoreceptorresponded to a steady dis-
A x h a n i c a ~ placement (shown by red trace) with APs at a rela-
displacement tively constant frequency throughout the stimula-
tion. (Top) Extracellularly recorded APs elicited by
three different amounts of disolacement, which in-
creased from bottom to top. The amount of dis-
placement is indicated below each record. (Bot-
tom) Steady-state frequency of APs plotted
against the number of grams of tension applied.
(B) Behavior of a phasic velocity detector. This
rapidly adapting mechanoreceptorresponded to
the rate at which the posltion changed. (Top)Ac-
tion potentials elicited by three different rates of
change (red traces) Higher velocities produced
more APs. (Bottom) The number of impulses pro-
duced during a 0.5 second stimulus was pro-
portional to the log of the veloclty of displace-
ment. [Adapted from Schmidt, 1971.1
I
1 0
I l l I L
0.2 0.5 1 2 5 10
Displacement velocity
Stimulus intensity (g) ( p m . ms-')
Stimulus Ca2+ can also activate Ca-dependent K+ channels,
I producing a shift in membrane potential, Vm, back
toward the resting potential.
5. The membrane of the spike-initiating zone (see
Figure 7-4) may become less excitable during sus-
tained stimuli.
6. Sensory adaptation can also take place in higher-order
cells in the central nervous system (which includes the
vertebrate retina).
Modulation of
3
of the CNS
Figure 7-12 Sensory adaptation can take place at any of several stages
in information processing. The dashed lines indicate that, in some sys-
tems, filtering or the modulation of AP frequency, or both, take place in
the receptor cell itself; whereas, in other systems, these functions take
place outside the receptor cell.
Crystal
\
'Ore
\
Myelin
&=-
A Stimulus
I
Figure 7-14 Adaptation in the Pacinian corpuscle depends on the me- terthe lamellae were removed, the neuron remained depolarized during
chanical properties of accessory structures. (A) Experimental arrange- most of the stimulus. (D) Receptor current flow in response to deforma-
ment for tapping a Pacinian corpuscle receptor with a piezoelectric tion at the sensory zone of the axon. The generator potential is con-
crystal-driven stylus. The electrical recording was made between the ducted electrotonically to the spike-initiating zone at the first node of
hook electrode on the axon and the oil-water interface. (B) Electrical re- Ranvier. If the generator potential is sufficiently large, it will bring the
sponse of the intact corpuscle. The neuron depolarized transiently at the sp~ke-initiatingzone to threshold, producing APs in the axon. [Adapted
onset and the offset of the stimulus (dashed red lines). (C) In contrast, af- from Loewenstein, 1960.1
A change in the filtering that is produced by accessory Mechanisms that enhance sensitivity
structures (mechanism 1)contributes importantly to the Many receptor cells produce APs-or release neurotrans-
rapid adaptation of the Pacinian corpuscle, a pressure and mitter independent of APs-spontaneously in the absence
vibration receptor found in the skin, muscles, mesentery, of stimuli. (The amount of transmitter released from non-
tendons, and joints of mammals (Figure7-14A).Each Pacin- spiking receptors varies with the membrane potential,
"ian corpuscle contains a region of receptor membrane that V,,, .)When these spontaneously active receptors are stim-
is sensitive to mechanical stimuli and that is surrounded by ulated, the frequency of their APs-or their non-spiking
concentric lamellae of connective tissue resembling the lay- release of transmitter-is increased or decreased above its
ers of an onion. When something presses on the corpuscle, baseline level. Several mechanisms enhance the sensitivity
deforming it, the disturbance is transmitted mechanically of receptors to sustained stimuli, and one important mech-
through the layers to the sensitive membrane of the receptor anism modifies the properties of the receptor's on-going
neuron. The receptor membrane normally responds with a spontaneous activity. The spontaneous release of trans-
brief, transient depolarization at both the onset and the off- mitter from receptor cells-whether it is mediated by APs
set of the deformation (Figure 7-14B). However, when the or by graded changes in Vm-has two important conse-
layers of the corpuscle are peeled away, permitting a me- quences. First, any small increase in stimulus energy will
chanical stimulus to be applied directly to the naked axon, produce an increase in the rate of firing above the spon-
the receptor potential obtained is sustained much longer, taneous level. Small receptor currents in response to weak
producing a more accurate representation of the stimulus stimuli modulate the impulse frequency by shortening the
(Figure 7-14C). Although the receptor potential still shows intervals between impulses (Figure 7-15). This modulation
some degree of adaptation (there is sag in the record shown of impulse frequency allows the receptors to be much
in Figure 7-14C), there is no distinctive response at the off- more sensitive to changes in stimuli than would be feasi-
set of the stimulus. The mechanical properties of the intact ble if the receptor current had to bring a completely qui-
corpuscle, which preferentially pass rapid changes in pres- escent spike-initiating zone to threshold. The input-output
sure, confer on the receptor neuron its normally phasic re- relations of such a sensory fiber are described by the sig-
sponse. This behavior explains, in part, why we quickly lose moid curve in Figure 7-16. In the unstimulated condition,
awareness of moderate, sustained pressures, such as the the firing frequency is already on the steep part of the
stimuli that wearing clothing produces on our skin. curve, so even a small input will produce a significant in-
Regardless of its site or mechanism of origin, adapta- crease in firing frequency.
tion plays a major role in extending the dynamic range of Second, in some spontaneously active sensory neurons,
sensory reception. Together with the logarithmic nature of stimuli can either increase or decrease impulse frequency,
the primary transduction process, sensory adaptation al- permitting the receptor to convey information about the
lows an animal to detect changes in stimulus energy against polarity or direction of a stimulus. For example, in some
background intensities that range over many orders of mechanoreceptors, such as hair cells, movement of the hair
magnitude. in one direction increases the rate of firing in the sensory
Stimulus Efferent control of receptor sensitivity
I decreases intervals The responsiveness of some sense organs is influenced by
the central nervous system through efferent axons that in-
nervate the sense organ itself. For example, the muscles to
which muscle stretch receptors in skeletal muscles of verte-
brates and crustaceans are attached are innervated by ef-
ferent fibers. By controlling the length of the receptor mus-
cles, this efferent innervation sets the sensitivity of the
stretch receptor to changes in overall muscle length.
I I I In crayfish and lobsters, when the tail extensor muscle
Time shortens, the receptor muscles (which run parallel to the
Figure 7-15 In a spontaneously firing receptor cell, the Interval between extensor muscle) shorten too, driven by efferent neurons.
APs depends on stimulus conditions. The interval can be decreased by If there were n o such mechanism, when the tail extensor
extremely small stimuli, because a stimulus increases the slope of the in- muscle shortened, the stretch receptors would go slack and
ternally generated depolarization. (In cases where APs are generated in would be unable to detect any further change in the length
second-order sensory fibers, synaptic potentlais, rather than internally of the extensor muscle. Instead, contraction of the recep-
generated depolarizations, are enhanced.)
tor muscles, in response to the efferent input, maintains a
fairly constant tension on the sensory parts of the receptor,
and the receptor retains its sensitivity to flexion of the tail,
regardless of the tail's position in space. In addition to this
mechanism by which the receptors maintain high sensitiv-
ity, the abdominal stretch receptors are innervated by ef-
Stin~ulusenergy
Figure 7-16 The input-output relation of a spontaneously active recep-
tor cell or second-order sensory fiber) is sigmoidal. In the absence of any
input (0 on the abscissa), the receptor cell, or second-order sensory fiber,
fires spontaneously This spontaneous output lies on the steep part of the
curve relating the stimulus intensity to the frequency of APs, so even a
very small stlmulus will increase the rate of flring. In some receptors, stim-
uli can also decrease the rate of firlng, indicated by the black dashed part
of the curve.
Activity
Activity
232 P H Y S I O L O G I C A L PROCESSES
...............................................................................
information about the chemical environment and trans- night from several miles downwind, an ability that confers
mitting it to other neurons. According to one widely ac- obvious reproductive advantage in a widely dispersed
cepted classification scheme, chemoreceptors can be species.
divided into two categories: gustatory (taste) receptors, To investigate the sensitivity of the receptors to bom-
which respond to dissolved molecules, and olfactory (smell) bykol, the electrical responses of antennal olfactory recep-
receptors, which respond to airborne molecules. This di- tors of Bombyx have been recorded. When only about 90
chotomy, however, rapidly breaks down. By these defini- bombykol molecules per second impinge on a single recep-
tions, aquatic organisms, such as fishes, could have no tor cell, the rate at which the cell fires APs increases signif-
smell receptors; all of their chemical sensation would be icantly. However, a male moth reacts behaviorally (e.g.,
taste. Furthermore, even in terrestrial organisms, airborne flaps his wings excitedly) when only about 40 receptor cells
molecules must pass through a layer of aqueous solution (out of a total of 20,000 per antenna) each intercept one
before they reach the olfactory receptors. molecule per sqcond. No change can be detected in the fre-
If taste and smell are legitimately different senses, there quency of APs fired by a single receptor cell in response to
must be a more useful distinction between them. Indeed, as a single odorant molecule. As a result, it is inferred that the
we will see, the receptors of taste and the receptors of smell moth's central nervous system is capable of sensing very
operate quite differently from one another, making it pos- slight average increases in impulse frequency arriving along
sible to distinguish between the two senses at the cellular numerous chemosensory channels, as described earlier in
level. In addition, there are alternative schemes to distin- this chapter (see Mechanisms that enhance sensitivity).
guish between taste and smell at a more global level. For
example, smell can be considered as the chemoreception of Mechanisms of Taste Reception
signals from distant sources and "taste" as the chemore- Electrophysiological studies of the contact chemoreceptors
ception of signals from material that is in direct contact (taste hairs) of insects have revealed generally useful infor-
with the receptive structure (e.g., masticated food in the mation about how chemoreception works. These receptor
mouth or material at the bottom of a pond in which a cat- cells send fine dendrites to the tips of hollow hairlike pro-
fish is living). jections of the cuticle, called sensilla (plural; singular, sen-
Chemosensory systems can be extraordinarily sensitive. sillum). Each sensillum has a minute pore that allows stim-
The antennal chemoreceptors of the male silkworm moth ulant molecules to reach the sensory cells (Figure 7-19). In
(Bombyx mori) for bombykol, the female's sex-attractant the proboscis or on the feet of an ordinary housefly, every
pheromone, provide a spectacular example. In the labora- sensillum contains several cells, each of which is sensitive to
tory, a male moth responds behaviorally to concentrations a different chemical stimulus (e.g., water, cations, anions,
of the pheromone that are as low as one molecule per loi7 or carbohydrates).
molecules of air. These receptors are highly specific, re- The electrical activity of these chemoreceptors can be
sponding only to bombykol and a few of its chemical recorded through a crack made in the wall of the sensillum,
analogs. This highly evolved stimulant-receptor system al- and such records have revealed both a receptor potential
lows the male Bombyx moth to locate a single female at and APs. The receptor potential is produced at the ends of
Insect olfactory
tor cells. All of these cells, or cell parts, that are regularly re-
receptor receptor
newed directly interact with physical stimuli from outside
the organism: taste and smell molecules in gustatory and ol- Figure 7-20 Chemosensoryorgans typ~callyconsist of receptor cells sur-
rounded by supporting structures. (A) In vertebrate taste buds, the re-
factory cells, respectively, and photons in photoreceptor
ceptor cells are surrounded by basal cells, which generate new receptor
outer segments. The turnover of all sensory cells poses a cells, and by supporting cells. Transduction takes place across the apical
problem for the maintenance of sensory specificity in an or- membrane. The receptor cells do not themselves send axons to the cen-
ganism because, unless the new cells were precisely inte- tral nervous system, although they can produce APs Instead, they synap-
grated into the existing network, specificity would be lost. tlcally exc~teafferent neurons that carry the ~nforrnat~on to the CNS In
contrast, vertebrate (B) and Insect (C) olfactory receptors themselves
Just how the integrity of taste and smell sensations is main-
send primary afferent axons to the CNS. Structures that are analagous be-
tained remains an unsolved, but actively studied, mystery. tween vertebrates and insects are drawn similarly in parts B and C. All
Although our subjective experience would suggest that three types of receptors extend fine processes into a mucous layer that
there is a wonderfully large spectrum of possible tastes, these covers the epithelium. In insects, these fine processes are true dendrites.
sensations can be grouped into four distinct qualities: sweet, [Part A adapted from Murray and Murray, 1970; part C adapted from
Steinbrecht, 1969.1
salt, sour, and bitter. In evolutionary terms, these categories
may be related to some basic properties of food. Sweet foods
are likely to be rich in calories and thus useful; salt is essen-
tial for maintaining water balance (see Chapter 14);a sour
taste can signal danger if it is in excess; and many bitter sub- How do molecules interact with membranes to pro-
stances are toxic. The discovery that vertebrates respond to duce distinct tastes? In the past few years, the use of patch-
only four fundamental categories of tastes suggests that all clamp recording has allowed the mechanisms responsible
perceived tastes must depend on various combinations of for each taste modality to be identified (Figure 7-21). Each
these fundamental characteristics. In addition, it generated individual taste receptor cell reacts to particular stimuli,
-the hypothesis that there is a separate, identifiable sensory and each class of taste stimuli activates a distinctive
pathway associated with each of the four tastes. cellular pathway in the receptors that respond to it. Salty
Figure 7-21 Each kind oftaste is transduced by a dist~nct~ve mechanism.
(A) In the transduction of salty and some sour tastes, Na+ (or H-) ions
pass through ion channels in the apical membrane, directly depolar~z~ng
the receptor cell (B) In the transduction of other sour tastes and some
b~ttertastes, protons (sour) or certain bitter compounds block Ki chan-
nels, and residual leakage of cations into the cell depolarizes the recep-
tor. (C) L-Alan~ne (Ala)and some other sweet compounds bind to recep-
tors (R) and actlvate a G protein (G).The activated G protein activates
adenylate cyclase (AC), and the resulting increase in cAMP closes K t
channels in the basolateral membrane, depolarizing the cell. (D) L-Argi-
nine (Arg)binds to, and opens, a ligand-gated, nonselective cation chan-
nel. (E) Some bitter compounds bind to a receptor and activate a G
protein that IS thought to be coupled to phospholipase C (PLC),produc-
ing an Increase In intracellular ~nositoltriphosphate (InsP,), which could
then release CaZ' from ~ntracellularstores. The net result 1s an increase in
transmitter release from the receptor cell, but the mechanism is not
yet completely understood. PIP,, phosphoinositol 4.5-biphosphate.
K+ [Adapted from Avenet et al., 1993.1
c / Ala and
4'sweet compounds
-n
i
Close K+
monosodiuin glutamate, activate nonspecific cation-selec-
tive channels in taste cells. Some bitter substances, such as
channels Ca2+ and quinine, close K + channels in the apical mem-
brane, allowing the cell to depolarize. Transduction of
other bitter substances is less well understood but appears
to rely on intracellular second-messenger systems (either
the InsP; or cAMP pathways) to excite the cell. It is hy-
pothesized that the sweet and bitter pathways that act
through second-messengers are mediated by G proteins,
and recent reports have suggested candidate molecules. In
all cases, the initial event in the receptor cell eventually
E B~ttercompound causes an increase in the concentration o f intracellular Ca'+
n n and thus increases the release of neurotransmitter onto the
second-order cells in the pathway.
Taste receptors generate APs, but they have no axons,
so they cannot themselves carry information to the central
nervous system. Instead, they synapse onto, and modulate
activity in, neurons whose axons run in the facial, glos-
G protein sopharyngeal, and vagus nerves (seventh, ninth, and tenth
cranial nerves). The existence of four kinds of taste sensa-
Ca2+stores tioils and the specificity of membrane transduction mech-
anisms for each kind of taste suggest that each receptor sub-
1
Ca2+
type might be connected to a particular set of axons. In that
arrangement, for example, information about "sweetness"
would be carried by some specific subset of axons. Such a
s t ~ m u l such
~ , as NaCI, read~lyd~ssociateIn water, and the pattern is called labeled line coding, but recordings have re-
N a + Ions enter receptors through N a + channels In the vealed that taste information is not nearly this neatly orga-
membrane to depolar~zethe membrane potent~al.These nized. Recordings from single neurons show that a recep-
Na+ channels are d ~ s t ~ n c t ~
because
ve they can be blocked tor will often respond optimally to a particular type of
by the drug a m ~ l o r ~ dunl~ke
e, the voltage-gated Na+ chan- stimulus (Figure 7-22), but many receptors also respond
nels that med~atemost APs. Sour s t ~ m u lw~ h, ~ c hare char- suboptimally to stimuli in other classes. The data thus sug-
acter~zedby excess H+ Ions, act e ~ t h e through
r t h ~ ssame gest that a single fiber innervating a taste bud receives in-
channel (observed in the hamster) or by blocking a K+ formation fromseceptors belonging t o different subtypes.
Chorda tympani nerve Figure 7-22 Each afferent taste neuron is
most effectively stimulated by one type of
stimulus, but it also responds to other stimuli.
The responses to four different taste stimuli
-
were recordedfrom sinqle taste afferent axons
in two different nerves in hamsters. Each neu-
ron responded maximally to one of the four
taste stimuli; different neurons responded
maximally to different stimuli. However, all of
the axons responded at least weakly to all four
stimuli, indicating that each taste afferent is
not restricted to carrying information about
only one kind of taste. Abbreviations: S, su-
m u
crose (sweet), N, NaCl (salty), H, HCI (sour), Q,
S N H Q S N H Q S N H Q S N H Q
-am-,
2 qulnlne HCI (b~tter)The number of axons IS In-
.-E Glossopharyngeal nerve d~catedfor each group [Adapted from
Hanamor~et al ,1988 ]
n=19
" S N H Q S N H Q S N H Q S N H Q
Stimuli
Rather than simple labeled line coding, sensory informa- flows over them during respiration (Figure 7-23).Animals
tion about taste must depend on the analysis of many gus- that are particularly dependent on olfactory cues have
tatory axons in parallel. complex cavities that are lined by sheets of receptors.
These cavities are called turbinates, and the mechanisms
Mechanisms of Olfactory Reception guaranteeing air flow through them remains unknown.
In vertebrates, olfactory receptors are located inside the Each receptor neuron has a long thin dendrite that termi-
nasal cavity, arranged so that a stream of air or water nates in a small knob at the surface (Figure 7-24A).
cilia ,
Solution aimed
at cilia
Solution aimed
at soma
Odorant Odorant
Emanating from the knob are several thin cilia (about transduction. The second piece of evidence comes from ex-
0.1 pm in diameter and about 200 pm long) that are cov- periments in which olfactory neurons were grown in cul-
ered by a protein solution called mucus. Molecules deliv- ture and were exposed to odorants while the receptor cur-
ered to the nasal cavity are absorbed into the mucous rent was recorded by an intracellular electrode in the soma
layer and delivered to the cilia. (Figure 7-24B). If a solution of odorant molecules was
Two lines of evidence suggest that the cilia are the lo- ejected onto the cilia, the cell responded strongly; whereas,
cation of olfactory transduction. First, only ciliated neurons if the same solution was ejected onto the soma, there was
respond to odors, implying that the cilia must be the site of only a small response. In contrast, ejecting a solution of
KC1 (which would depolarize the membrane of the recep- contrast with the small number of receptor types that code
tor) onto the cilia produced a small response, whereas eject- for taste.
ing KC1 onto the soma produced a large response. These Olfactory coding in vertebrates has been studied
data imply that only the cilia were able to respond to the electrically in the olfactory epithelium of the frog
odorant, causing V,,, to change significantly. (Figure 7-25A) In these experiments, activity of single
The olfactory transduction cascade includes an ade- receptor axons was recorded by one electrode while
nylate cyclase that is linked to a G protein. (Seethe discus- the summed potential of large numbers of olfactory recep-
sion of transduction earlier in the chapter.) A very large tors in the epithelium (the electro-olfactogram, or EOG)
family of proteins has recently been found to be expressed were recorded simultaneously by another electrode
only in olfactory epithelial cells. The structure of each pro- (Figure 7-25B). Impulses from individual receptors
tein includes seven transmembrane domains, and other fea- were then superimposed electronically on the electro-
tures also indicate that these molecules are homologous to olfactogram. This technique permits the activity in a single
proteins that mediate other transduction processes. The receptor to be compared with the total response of many
large size of this family of proteins suggests that there could receptors when a single odorant, or a combination of odor-
be many individual receptor subtypes for distinct odors, in ants, is presented.
I
Moist,
Odors
cleaned
air
B
EOG
Electrode 1 7
Electrode~
=~,+'2/
Mucous
layer\
Olfactory
cilia
L ' - Olfactory
cell
olfactory
nerve
The results indicate that stimulus coding in the ver- between classes of odorants and types of olfactory cells
tebrate nose is far more complex than is coding in the con- in the frog. Instead, each olfactory receptor cell appears
tact chemoreceptors of houseflies. Different receptors re- to express a mosaic of odorant receptor molecules with
spond differently to the same odorant. In some olfactory differing specificities. The response characteristics of a
axons, a particular odorant increased the impulse fre- particular olfactory receptor must, then, depend on the
quency (Figure 7-26A). Some odorants that smell alike to proportions of its many types of receptor molecules. This
humans have similar effects on some frog olfactory cells, situation implies that the ability of mammals to distin-
suggesting that they would smell alike to frogs, too. How- guish among a wide variety of odors must reside in the
ever, these same odorants have different effects on other ability of the higher olfactory centers in the brain to de-
cells (see Figure 7-26A, cell a vs. cell b), suggesting that code a combinatorial signal from a large number of olfac-
they would not smell alike to the frog. In the olfactory tory receptors.
bulb, farther along the chain of olfactory neurons, neu-
rons may respond to an odorant with decreased activity
MECHANORECEPTION
-
or with increased activity (Figure 7-26B). In fact, it has
proved to be impossible to establish a one-to-one relation All animals can sense physical contact on the surface of
their bodies. Such stimuli are detected by mechanorecep-
tors, the simplest of which consist of morphologically un-
Resting discharge
differentiated nerve endings found in the connective tissue
of skin. More complex mechanoreceptors have accessory
Menthone
structures that transfer mechanical energy to the receptive
membrane. These accessory structures often also filter the
mechanical energy in some way, as described earlier in re-
Menthol gard to the mammalian Pacinian corpuscle in which the
sensitive ending is covered by a capsule (see Figure 7-14).
Other mechanoreceptors include the muscle stretch recep-
tors of various kinds found in arthropods and vertebrates,
Cell a Cell b
in which mechanically sensitive sensory endings are associ-
ated with specialized muscle fibers (see Figure 7-13), and
Odorant applied the hairlike sensilla that extend from the exoskeletons of
arthropods (Figure 7-27). The most elaborate accessory
structures associated with mechanoreceptive cells are found
in the vertebrate middle and inner ear and in the vestibu-
lar system, both of which are considered later in this
chapter.
The stimulus that activates mechanoreceptive mem-
brane is a stretch or distortion of the surface membrane.
Suppression
of ongoing
Indeed, stretch-sensitivechannels are found in all types of
organisms from the simplest to the most complex. Patch-
clamp data indicate that these channels respond to changes
in tension in the plane of the membrane, and they can be ei-
ther activated or inactivated by stretch. Stretch-sensitive
channels defy simple classificationwith regard to selectiv-
Increased
activity I ity, because they show a wide range of conductances and fi-
delity. Possible transducers of mechanical stress include the
cytoskeleton, enzymes, or the ion channels themselves. Me-
chanically sensitive channels are the only primary mechan-
Figure 7-26 Olfactory receptors have complicated responses to indi- otransducers that do not depend on enzymatic activity but
vidual odorants. (A) Recordings from two frog olfactory receptors. Men- instead directly use the free energy stored in the transmem-
thone and menthol both slightly suppresssed ongoing activity in cell a,
brane electrochemical gradient. Mechanoreceptors can be
indicating that cell a could not distinguish between the two substances.
In contrast, cell b responded differently to the two substances, produc- exquisitely sensitive, responding to mechanical displace-
ing more APs in response to menthol than it did at rest but fewer in re- ments of as little as 0.1 nm. It is a continuing challenge to
sponse to menthone than it did at rest. Thus, cell b could potentially dis- understand how such small displacements can produce
tinguish between the two substances, whereas cell a could not. Notice changes in ion permeability through the membrane.
that the electro-olfactogram(EOG) has been summed with the individ-
ual record for each cell. (B) Recordingsfrom second-order olfactory cells Hair Cells
of the tiger salamander. Odorants may reduce or increase ongoing ac-
tivity in these cells. [PartA adapted from Gesteland, 1966; part B adapted The hair cells of vertebrates are extraordinarily sensitive
from Kauer, 1987.1 mechanoreceptors that are responsible for transducing me-
SENSING T H E E N V I R O N M E N T 239
.............................................
Hair plate
w
-/
Hair plate'
;I-
~rochanter
Femur
air plate
chanical stimuli into electrical signals (Figure 7-28). They stereocilia, making a hair cell bilaterally symmetric, with
are found in several locations. Fish and amphibians have a the top beveled like a hypodermic needle. In most organs,
set of external receptors, called the lateral-line system, that the hair bundles are coupled to some kind of accessory
are based on hair cells and that detect motion in the sur- structure through their kinocilia. Stimuli that affect the
rounding water (Figure 7-29). The vertebrate organs of accessory structure are transmitted to bundles of stereo-
hearing and the organs that report the position of the body cilia through bonds that connect the accessory structures
with respect to gravity (called the organs of equilibrium) and kinocilium to the stereocilia. In addition, if the tip
also are based on hair cells. Typically, the organs of equi- of the bundle of stereocilia is touched with a fine probe,
librium include the semicircular canals and the vestibular the bundle moves as a unit, regardless of the direction of
apparatus. stimulation.
Hair cells are named for the many cilia that project The exact process by which pressure or force from the
from the apical end of each cell. These cilia fall into outside world moves bundles of stereocilia depends on the
two classes: each hair cell typically has a single kinocil- specific arrangement of hair cells and accessory structures
ium and 20-300 nonmotile stereocilia. The kinocilium within each sense organ, but ultimately it is the movement
has a "9 + 2" arrangement of internal microtubules (see of the stereocilia that produces an electrical signal. When
Figure 7-28A) that are similar to that of other motile cilia. the cilia bend toward the tallest cilium, a hair cell depolar-
The stereocilia contain many fine longitudinal actin fila- izes; whereas, when they bend in the opposite direction, the
ments, and they are thought to be both structurally and de- cell hyperpolarizes (see Figure 7-28D). (If the stereocilia
velopmentally distinct from the kinocilium. bend to either side, rather than toward or away from the
Although the hair cells of the lateral line and of the or- kinocilium, Vm remains the same.)
gans of equilibrium have both a kinocilium and several At rest, about 15% of the channels in a hair cell are
stereocilia, some hair cells in the adult mammalian ear lack open, producing a resting potential of about -60 mV. Hair
kinocilia. In addition, the technically remarkable feat of mi- cells do not produce APs. Instead, they form chemical
crosurgically removing the kinocilia from hair cells that synapses onto afferent neurons and release neurotransmit-
normally have them does not block transduction. From ter in a graded fashion, depending on V, in the receptor
these two observations, it appears that kinocilia must not neuron; the afferent neurons then carry information into
be necessary for mechanotransduction. The stereocilia of a the central nervous system. The amount of transmitter re-
hair cell are arranged in order of increasing length from one leased onto the afferent neurons determines their frequency
side of the cell to the other (see Figure 7-28B and C). A of discharge. Notice that the input-output relation for hair
plane of symmetry through the kinocilium bisects the cells is markedly asymmetrical (see Figure 7-28D); that is,
240 PHYSIOLOGICAL PROCESSES
...............................................................................
-C3
lnh~b~t~on
e+
Exc~tation
St~mulusmovements
+
-
lntracellular receptor potent~al
I H a ~ rcell
!L
iP Nerve ~mpulses
Displacement (prn)
SENSING THE ENVIRONMENT 241
........................................
Figure 7-28 (At left) The membrane potential of hair cell receptors the depolarization produced by a given amount of dis-
changes when the cilia are moved away from a rest position. (A) Electron placement toward the tallest cilium is larger than the hy-
micrograph of a cross sect~onthrough the cilia of a hair cell. The large cil-
+
ium, containing a typical 9 Zstructure of microtubules, is the kinocil-
perpolarization produced by a similar displacement of the
ium; the others are stereocilia. (B) Scanning electron micrograph show- cilia in the opposite direction. This asymmetry is important
ing the structure of a hair cell from the neuromast of a giant Danio fish. because, when hair cells are subjected to symmetrical vi-
(C)Diagram of a typical hair cell showing the anatomical relations of the brations such as sound waves, changes in the membrane
stereocilia and the kinocilium The hair cell releases transmitter onto an af- potential can faithfully follow the alternating phases of the
ferent neuron, which carries the sensory signal t o the central nervous sys-
tem. It also receives synapses from efferent neurons. Depending on the
stimulus only up to frequencies of several hundred hertz
direction in which the cilia are bent, the hair cell can either increase or de- (Hz)but sound frequencies are often much higher than this
crease the frequency of APs in the afferent fiber. A linear back-and-forth value. At higher frequencies, the response to the vibrations
motion applied t o the cilia produces intracellular potential changes that fuses into a steady depolarization; even if the stimulus dis-
can b e recorded with a microelectrode. Extracellular recording from the places the cilia in both directions by equal amounts away
afferent axon shows APs associated with changes in Vm in the receptor
cell. (D) Input-output relation for a hair cell. Note that the depolarization
from zero displacement, the hair cell will depolarize. This
produced by movementtoward the kinocilium is largerthan the hyper- steady depolarization in response to high frequency stimuli
polarization in response t o movement away from the kinocilium. [PartA produces steady, rather than modulated, transmitter release
from Flock, 1967; part B courtesy of Christopher Braun; part C adapted by the hair cell and, hence, high frequency firing of the af-
from Harris and Flock, 1967; part D adapted from Russell, 1980.1 ferent neurons. The details of transduction by hair cells are
presented later (see Excitation of cochlear hair cells).
Organs of Equilibrium
The simplest organ that has evolved to detect an animal's
position with respect to gravity or its acceleration is the
statocyst. Forms of this type of organ are found in a num-
ber of animal groups, ranging from jellyfish to vertebrates.
(Interestingly, insects lack these sense organs and appar-
ently depend entirely on other senses, such as vision or joint
proprioceptors, for orientational information.) A statocyst
consists of a hollow cavity lined with ciliated mechanore-
ceptor cells that make contact with a statolith, which can
be sand grains, calcareous concretions, or some other rel-
atively dense material (Figure7-30A). The statolith is either
taken up from the animal's surroundings or secreted by the
epithelium of the statocyst. For example, a lobster loses its
statoliths at every molt and replaces them with new grains
airs embedded of sand. In either case, the statolith must have a higher spe-
cific gravity than the surrounding fluid.
As the position of the animal changes, the statolith rests
on different regions of the statocyst. When a lobster is tilted
to the right about its longitudinal axis, the statolith rests on
the receptor cells on the right side of the statocyst, stimu-
lating them and causlng a tonic discharge in the sensory
fibers from the stimulated receptor cells (Figure 7-30B).
Recordings from many different fibers of a statocyst reveal
-
that each cell fires maximally in response to a certain ori-
entation of the lobster (Figure 7-30C). Information from
these receptors travels to the central nervous system and
sets up reflex movements of the appendages. This pattern
of information processing was confirmed in a clever exper-
iment in which molting lobsters were presented with iron
Figure 7-29 The lateral-line sensory system of fish and amphibians is filings, rather than with sand. They replaced their statoliths
based on hair cells. The drawing shows the location of these receptive or- with iron filings, allowing the position of the iron statolith
gans along the body of an African clawed frog, Xenopus. The lower dia-
to be manipulated by a magnet. As the magnet was moved
gram shows a cross-section through part of the lateral line, illustrating the
cupula, an accessory structure that bends cilia when it is displaced by mo- through space, pulling on the iron statolith, the lobster-
tion of the surrounding water. Compare the structure of this organ with whose position with respect to gravity had not changed-
the hair cells shown in Figure 7-28. produced a series of compensatory postural responses.
242 P H Y S I O L O G I C A L PROCESSES
...............................................................................
nule
0 20 40 60
Degrees
Figure 7-30 Statocysts sense accelerat~onand the pos~t~on of an an~rnal trace below each recording indicates the time course of the tilt and the
w ~ t hrespectto grav~ty(A) Structure of a statocyst In a lobster Astatol~th angle to which the animal was tilted. (C) Frequencies of APs recorded
rests on the receptor part of an array of ha~rcells (B) Act~onpotent~als from different fibers plotted as a function of the position of the animal.
recorded from ~nd~v~dually d~ssectednerve f~berswh~lethe lobster was Each cell responded with a maximum rate of discharge at a different po-
tllted Each recording shown here was made from a d~fferentf~berThe sition. [Adapted from Horridge, 1968.1
The Vertebrate Ear fluid moves in one direction and inhibited when it moves in
The ears of vertebrates perform two sensory functions, the opposite direction. The orthogonal arrangement of the
each of which is based on the activity of hair cells. Some three canals allows them to detect any movement of the
structures of the ear, the organs of equilibrium, perform like head in three-dimensional space.
the statocystsof invertebrates, reporting on the animal's po- Below the semicircular canals, larger bony chambers
sition with respect to gravity and acceleration through contain three more patches of hair cells called maculae.
space. Other structures, the organs of hearing, provide in- Mineralized concretions termed otoliths are associated with
formation about vibrational stimuli in the environment- the maculae, similar to the statoliths associated with stato-
stimuli that are called sound if they fall within a particular cysts. The otoliths signal position relative to the direction
frequency range. of gravity; in the lower vertebrates, they can also detect vi-
brations, such as sound waves, in the surrounding medium.
Vertebrateorgans of equilibrium Sensory signals from the semicircular canals are integrated
In vertebrates, the organs of equilibrium reside in a mem- with other sensory input in the brain stem and in the cere-
branous labyrinth that develops from the anterior end of bellum for the control of postural and other motor reflexes.
the lateral-line system. It consists of two chambers, the sac-
culus and the utriculus, that are surrounded by bone and The mammalian ear
filled with endolymph, a specialized fluid. Endolymph dif- Sound in the environment has led to the evolution of hear-
fers from most extracellular fluids because it is high in K+ ing in many phyla. Hearing allows an animal to detect
(about 150 mM in human beings) and low in Na+ (about predators or prey and to estimate their location and dis-
1 mM in human beings); the significance of this unusual tance while they are still relatively far away. Sound also
composition is considered in the section titled The mam- plays an important role in intraspecific acoustic communi-
malian ear. The utriculus gives rise to the three semicircular cation, which often requires subtle tuning of both produc-
canals of the inner ear, which lie in three mutually perpen- tion and reception. Sound is a mechanical vibration that
dicular planes (Figure 7-31). Hair cells in the three orthog- propagates through air or water, traveling as waves of al-
onal semicircular canals detect acceleration of the head. As ternating high and low pressure, accompanied by a back-
the head is accelerated in one of the planes of the canals, the and-forth movement of the medium in the direction of
inertia of the endolymphatic fluid in the corresponding propagation. The nature of sound, particularly the differ-
canal ~roducesa relative motion of the endolymph past a ences in how it is conducted through air and through wa-
gelatinous projection, the cupula, which moves the cupula. ter, has set distinct constraints on its detection. The evolu-
When the cupula moves with respect to the cilia of the hair tion of hearing illustrates many different mechanisms that
cells at its base, V, of the hair cells changes. All hair cells in have evolved to solve the various problems presented by the
the canal are oriented with the klnocilium on the same side, physical nature of sound. A well-studied example, which
so all hair cells attached to the cupula are excited when the we examine here, is the mammalian ear.
SENSING THE E N V I R O N M E N T 243
...............................................................................
B
Superior
Tem~oralbone
.. .
. __-- r/ Endolymphatic
duct
canals Endolymphatic
/
Cochlear
nerve (VIII)
Incus I
Tragus
'External
auditory
canal
in oval
window > ,
Eustachian ,
Scala
media
tube I
I window
; Cochlea
\\ Scala
I
I vestibuli
,,
I
I
C Otoconia
,
I I
i
;
'
I
,Gelatinous mass
of cupula
\ Gelatinous
layer
1
;
fiber
Figure 7-31 The human auditory organs and organs of equilibrium are ceptors in a semicircular canal are embedded in the gelatinous cupula.
located in the ear. (A) The major parts of the ear. (B) The semicircular When fluid moves in the canal, the cupula bends the cilia (left). Particles
canals and cochlea. Thestapes has been removedto reveal the oval win- called otoconia rest on the cilia of receptors in the sacculus (one of the
dow. The pathway taken by auditory signals is shown by black arrows. At maculae).Changes in the position of the head cause the otoconia to shift
the far right, a section has been removed from the cochlea to reveal the posit~on,changing how much the cilia are bent (right). [Parts A and B
inner structure. (Figure 7-33 shows this structure in more detail.) (C) De- adapted from Beck, 1971; part C adapted from Williams et al., 1995.1
tailed structure of two parts of the organs of equilibrium. The cilia of re-
External ear, auditory canal, and middle ear The structure To be detected, airborne vibrations must be transmitted
of the external ear acts as a funnel that collects sound waves to the fluid-filled inner ear, where the receptor hair cells re-
in the air from a large area and concentrates the oscillating side. The difficulty of communicating across a air-liquid in-
air pressure onto a specialized surface, the eardrum or tym- terface can be appreciated by trying to talk with someone
panic membrane. The external structures of the ear-the who is under water. Most of the sound energy generated in
pinna and tragus-facilitate the collection of sound waves. air is reflected back from the water's surface, so it is difficult
The shell-like outer structures, and in some species the mo- to generate sufficient energy with airborne sounds to move
bility of the shell, can modify the directional sensitivity of the water at the required frequency and displacement. This
the auditory system. In some species, including human be- kind of situation is called acoustical impedance mismatch.
ings, the acoustical properties of the external ear amplify In the ear, this mismatch is partially overcome by a series of
sound in particular frequency ranges. In addition, the hu- three small bones connected in series that are attached to
man ear emphasizes the spatial distribution of stimuli by the tympanic membrane at one end and to the oval window
amplifying sounds coming from some directions more than of the cochlea at the other. These bones, the auditory ossi-
it does sounds from other directions (Figure 7-32). cles (labeled incus, malleus, and stapes in Figure 7-31A),
Figure 7-32 The structure of the human
plnna and tragus selectively a m p l ~ f ~ espe-
s
clf~cfrequenc~es of sound Thls graph shows
the galn In pressure at the eardrum over
what the pressure would b e ~f sound Irn-
plnged on the ear canal w ~ t hall external ear
structures removed If there were n o ampll-
f~cat~on, the graph would be a hor~zontall ~ n e
lntersectlng w ~ t hthe ordinate at an arnpltfi-
c a t ~ o nfactor of 1 Values above 1 lnd~cate
evolved from the articulation points of the posterlor jaw Figure 7-31A and B). It is divided internally into three lon-
and are now located in the middle ear. Changes In air pres- gitudinal compartments (see Figure 7-33A). The two outer
sure produced by sound waves in the external auditory compartments (scala tympani and scala vestibuli) are con-
canal cause the eardrum to move, whlch transfers the en- nected by the helicotrema, an opening located at the api-
ergy first to the ossicles, and then to the structures of the in- cal end of the cochlea (see Figure 7-35B).The scala tympani
ner ear. In the inner ear, the first structure to receive the me- and scala vestibuli are filled with an aqueous fluid called
chanical input is the oval window, which forms the the perilymph, which resembles other extracellular fluids
outermost surface of a fluld-filled chamber (the cochlea) in having a relatively high concentration of Na+ (about
that contains the receptor hair cells. At the other end of the 140 mM) and a low concentration of K+ (about 7 mM).
fluid-filled compartment is another membrane, the round Between these compartments-and bounded by the basi-
window. lar membrane and Reissner's membrane-is another com-
There are two important consequences of this arrange- partment, the scala media, which is filled with endolymph
ment. First, the properties of the mechanical coupling be- (high in K+ and low in Na+),the same type of fluid that sur-
tween the eardrum, ossicles, and oval window amplify the rounds the cilia of hair cells in the organs of equilibrium.
signal by about 1.3 times. Second, the pressure of the signal The unusual ionic composition of endolymph contributes
is greatly amplified between the eardrum and the oval win- importantly to the process of auditory transduction. The
dow because the eardrum has an area of about 0.6 cm2, organ of Corti, which bears the hair cells that transduce au-
whereas the oval wlndow is smaller, about 0.032 cm2.This ditory stimuli into sensory signals, lies within the scala me-
ratio of about 17: 1 between the areas of the two mem- dia and sits on the basilar membrane, and signal transduc-
branes means that the sound pressure onto the typanum is tion by the cochlear hair cells depends in part on this
concentrated onto the smaller area of the oval wlndow, anatomical arrangement.
producing a much greater pressure, whlch IS important be- Among the vertebrates, only mammals possess a true
cause the inertia of the cochlear fluid on the other side of cochlea, although birds and crocodilians have a nearly
the oval window is greater than that of alr. The increase in straight cochlear duct that contains some of the same fea-
pressure helps to efficiently transfer airborne vibrations to tures, including the basilar membrane and the organ of
the cochlear fluld. As a consequence of these two mechan- Corti. The other vertebrates have no cochlea. Some of the
ical features, signals arriving at the eardrum are amplified lower vertebrates are able to detect sound waves through
by at least a factor of 22 by the tlme they reach the cochlea. the activity of hair cells associated with the otoliths of the
utriculus and sacculus and with the lagena, one of the three
Structure and function of the cochlea This mechanically maculae in the organs of equilibrium.
amplified sound input is transduced into neuronal signals The hair cells of the mammalian cochlea encode both
by the hair cells of the inner ear. The hair cells of the mam- the frequency (i.e., the pitch) and the intensity of sound.
malian ear are located in the organ of Corti in the cochlea The adult human cochlea contains four rows of hair cells,
(Figure 7-33). The movement of fluid in the cochlea causes one inner and three outer rows, with about 4000 hair cells
a vibration of the hair cells, which displaces their stere- in each row (see Figure 7-33B). The stereocilia of the hair
ocilia; the hair cells, in turn, excite the sensory axons of the cells contact the overlying tectorial membrane. The cilia are
auditory nerve. The hair cells in the organ of Corti resem- bent by shearing forces (i.e., a force perpendicular to the
ble the hair cells of the lateral-line system in lower verte- axis of the cilia) that arise when the hairs move through the
brates, except that the kinocilium is absent from some gelatinous mucus that coats the tectorial membrane.
cochlear hair cells in adults. Sound vibrations are transferred by the ossicles to the
The cochlea, a tapered tube encased in the mastoid oval window and then pass through the cochlear fluids and
bone, is coiled somewhat like the shell of a snail (see the membranes that separate the cochlear compartments
SENSING THE ENVIRONMENT 245
...............................................................................
A
Reissner's membrane
\
Spiral
glnnlinn
U
Rods of Corti membrane
\Afferent and efferent
nerve fibers
Figure 7-33 Sound stimuli are transduced by hair cells in the cochlea. basilar membrane in the central canal. (B) Enlargement of the organ of
(A) Cross section through cochlear canal, made at about the location il- Corti. The cilia of the hair cells are embedded in the gelatinous layer of
lustrated in Figure 7-31B, showing the two outer chambers (the scala the tectorial membrane, whereas their cell bodies are fixed with respect
vestibuli and the scala tympani) and the organ of Corti attached to the to the basilar membrane.
(both Reissner's membrane and the basilar membrane) be- imagine a displacement of the eardrum transferred through
fore their energy is dissipated through the membrane-cov- the ossicles of the middle ear to the oval window. Vibra-
ered round window. The distensibility of the round and tions displace the incompressible perilymph along the scala
oval windows is an important adaptation because, if the vestibuli, through the helicotrema, and back through the
fluid-filledcochlea were encased entirely by solid bone, the scala tympani toward the round window.
displacements of the oval window, the fluid, and the inter-
nal tissues would be very small. The distribution of the per- Excitation of cochlear hair cells Electrical recordings made
turbations within the cochlea depends on the frequencies of at various locations in the cochlea show fluctuations in elec-
vibrations entering the oval window. To visualize this, trical potential that are similar in frequency, phase, and
246 PHYSIOLOGICAL PROCESSES
.........................................
amplitude to the sound waves that produced them. These mechanical influence on the conformational states of the
cochlear microphonics.result from the summation of re- channel.
ceptor currents from the numerous hair cells that were Several factors affect the sensitivity of hair cells. Each
stimulated by movements of the basilar membrane. The ac- hair cell in the cochlea appears to be tuned to a particular
tual transduction event occurs when a perturbation of the band of sound frequency as a result of both mechanical and
basilar membrane forces the tips of the stereocilia to bend channel properties. Each cell has a resonance frequency
laterally, because the basilar membrane has moved with re- that is determined by the length of the stereocilia in the hair
spect to the tectorial membrane (Figure 7-34). This me- bundle. Cells with long hairs are most sensitive to low-
chanical deflection directly causes ion channels in the tips frequency sounds, whereas cells with short hairs are tuned
of the stereocilia to open. In the past few years, our under- to high-frequency sounds. In addition, each cell responds
standing of these events has grown dramatically, although maximally to a particular frequency of electrical stimula-
questions still remain about the details of transduction. tion. This electrical resonance frequency is determined by
The perceptual threshold of cochlear hair cells cor- the balance of currents through voltage-gated Ca2+chan-
responds to a deflection of 0.1-1.0 nm, which is equiva- nels and through Ca2+-sensitiveK+ channels in the basal
lent to a change in membrane current of only about 1 pA membrane (which is exposed to perilymph).
through ion channels in the hair cell membrane. These The outer hair cells of the cochlea may contribute to
channels have been experimentally shown to be permeable tuning in the cochlea by modifying the mechanical proper-
to many small, monovalent cations (e.g., Li+, Na+, K+, ties of the organ of Corti. The outer hair cells make-few af-
Rb+, and Cs+).When they open in uiuo, K+ ions and some ferent connections, but they receive a large number of ef-
Ca2+ ions enter the cell from the endolymph. (The high ferent synapses. When these cells are stimulated electrically
concentration of K+ in the endolymph produces a net in- during experiments, they shorten when depolarized and
ward driving force on K+, unlike the usual situation in elongate when hyperpolarized. Thus, it is possible that the
which V, - E, is an outward force. This inward K+ cur- outer hair cells could modify the mechanical coupling be-
rent depolarizes hair cells, because it adds positive charge tween the inner hair cells and the tectorial membrane,
to the inside of the cells.) which would change transduction. It remains to be demon-
On the basis of measurements of current flow, it has strated that this mechanism actually affects audition.
been estimated that there are about 30-300 channels per Hair cells adapt to changes in the position of their stere-
bundle of stereocilia, which implies that as few as one to ocilia, a process that has been particularly well studied in
five channels per stereocilium may be responsible for trans- the bullfrog sacculus. When the cilia of frog hair cells are
duction. The channels are thought to be opened directly by deflected by a probe and held at the new position, the op-
a mechanical stimulus because, when isolated bundles of erating range of the cell adapts within milliseconds to this
stereocilia are abruptly deflected in experiments, the trans- new tonic position, causing the hair cell to then respond to
duction current increases with an extremely short latency small changes in position away from this new set point.
(about 40 ps). The brevity of this latency period makes it Calcium ions have been shown to play a pivotal role in the
unlikely that an enzymatic or biochemical step is included process, evidently by modifying the tension in the spring
in this process. This interpretation is reinforced by patch- that opens the transduction channels. Finally, efferent input
clamp experiments indicating that the channels open faster onto a hair cell can decrease the cell's response to sound
when the deflection is larger, again suggesting a direct and broaden its frequency selectivity by opening inhibitory
Hinge
Tectorial
point
membrane
deflected up
hair cells membrane
hair cell point
Figure 7-34 Movement of the bas~larmembrane w ~ t hrespect t o the polnts when they are d~splacedby waves travel~ngalong the cochlea The
tector~almembrane produces shear on the stereoc~l~a of cochlear ha~r movements are greatly exaggerated In thls d~agram[Adapted from
cells The tector~almembrane sl~desover the organ of Cort~,because the Dav~s,1968 ]
tector~almembrane and the bas~larmembrane p~votabout d~fferent
SENSING THE E N V I R O N M E N T 247
......................................
K+ channels, which short-circuit the cell's electrical reso- varies in an orderly fashion with higher frequencies dis-
nance. Taken togetheq the attributes of the hair cells reveal placing the basilar membrane close to the oval window and
their exquisite tuning. However all of the adaptations that lower frequencies displacing the basilar membrane farthest
make hair cells so extremely sensitive also make them from the oval window. For sounds up to about 1kHz, APs
highly vulnerable to overstimulation, which can cause rup- in the auditory sensory axons appear to follow the funda-
ture at the base of the stereocilia. Acoustic trauma can pro- mental frequency. Above this level, the time constant of the
duce permanent hearing loss that is worst at the frequencies hair cells and the electrical properties of the axons in the au-
of sound that actually damaged the hair cells. Although ditory nerve prevent a one-to-one correspondence between
some cold-blooded vertebrates can recover from such sound waves and electrical signals. In this higher frequency
trauma, the loss is permanent in mammals. range, some other mechanism must inform the central ner-
vous system of the sound frequency.
Hermann von Helmholtz noted in 1867 that the basi-
lar membrane consists of many transverse bands that in-
crease gradually in length from the proximal end to the api-
cal end of the basilar membrane (from about 100 pm long
at the base to about 500 pm long at the apex), which re-
minded him of the strings of a piano and led him to pro-
pose his resonance theory. He proposed that various loca-
tions along the basilar membrane vibrate in resonance with
a specific tonal frequency while the other locations remain
The receptor currents of the hair cells faithfully trans- stationary, just as the appropriate string of a piano res-
duce the movements of the basilar membrane over the onates in response to a tone from a tuning fork. This
whole spectrum of audible sound frequencies. The cells theory was later challenged by von Btktsy (1960), who
transmit their excitation through chemical synapses onto found that the movements of the basilar membrane are not
sensory axons of auditory neurons that have cell bodies standing waves, as Helmholtz suggested, but consist in-
located in the spiral ganglion. Release of neurotransmitter stead of traveling waves that move from the narrow base
by the hair cells modulates the firing rate of these axons, of the basilar membrane toward the wider apical end
which travel in the uestibulocochlear (eighth cranial) nerve (Figure 7-35). These waves have the same frequency as the
and synapse onto neurons in the cochlear nucleus. In fact, sound entering the ear, but they move much more slowly
the hair cells in the inner row receive about 90% of the than sound moves through air.
contacts made by neurons of the spiral ganglion, suggest- A familiar example of a traveling wave can be seen by
ing that the inner row of cells is largely responsible for moving the free end of a rope that is secured at the other
detecting sound. In contrast, hair cells of the outer three end. Unlike a rope, however, the basilar membrane has me-
rows receive many efferent synapses and may participate chanical properties that change along its length. The com-
in modulating the sensitivity of the cochlea by changing pliance of the membrane (the amount that the membrane
the mechanical relation between the basilar and tectorial will stretch in response to a given amount of force) increases
membranes. from its narrow end to its broad end, which causes the am-
plitude of a traveling wave to change along the length of the
Frequency analysis by the cochlea Pioneering studies of the membrane (see Figure 7-35). The position along the cochlea
exposed cochlea, carried out by Georg von Btktsy, greatly at which the displacement of the basilar membrane is max-
enhanced our understanding of how the auditory system imal-causing the hair cells at that location to be stimu-
can encode information about the frequency of stimuli. His lated maximally-depends on the frequency of the travel-
studies showed that: ing waves and consequently on the frequency of the
stimulating sound. The traveling waves produce maximum
1. In response to a pure sine wave tone, the perturbations displacements near the basal end of the cochlea when the
of the basilar membrane have the same frequency as stimulating sound has a high frequency. The region of max-
the tone. imal displacement moves along the basilar membrane to-
2. Low frequency perturbations move as a traveling wave ward the apex as the frequency of the sound drops. The ex-
along the whole length of the basilar membrane. tent of membrane displacement at any point along the
3. The location where the basilar membrane is displaced basilar membrane determines how strongly the hair cells are
maximally by a tone is a function of the frequency of stimulated, so it also determines the rate of discharge in sen-
the tone. High frequencies displace only the initial sory fibers arising from different parts of the basilar mem-
parts of the membrane, whereas low frequencies dis- brane. Even at maximum amplitude, all of the movements
place more distant parts. are very tiny: the loudest sounds produce displacements of
the basilar membrane of only about 1pm. The movement
Thus, each point along the basilar membrane is displaced of the hair cell cilia is much smaller, and the threshold of
most effectively by some unique frequency, and that point stimulus detection is at the lirmt of the thermal noise.
20
Basal
end
- 22 24 26 28
Distance from stapes (rnm)
30
- Apical
end
Figure 7-36 A cricket's ears are located on its anterior thoracic legs. The
tympanum receives sound stimuli through the tracheal ducts and vibrates
in response either t o sounds from the outside or to sounds carried
Oval Basilar membrane Apex through the inside of the animal through the tracheae. Nerve cells asso-
ciated with the tympanum transduce the sound stimuli.
orsal branch
Receptor areas
-
Stimulus current
B Current
I I
Receptor potential
Y
- Resting
"
-1 0 +1
Stimulus voltage (mV)
Figure 7-37 Electroreceptorcells are specialized hair cells located along cules spontaneously (a) Current enter~ngthe cell (b)depolar~zes~ tIn- ,
the lateral line of many species of fishes. (A) Positions of the electric or- creasing the rate of release and, hence, the frequency of APs In the sen-
gan and of the lateral-linenerve trunk and distribution of electroreceptor sory f~ber~nnervatlngthe cell Current leav~ngthe cell (c) decreases the
pores in the weakly electric fish Gnathonernus peters;;. (B) At the base rate of release The amount of transm~tterreleased by the receptor cells
of each electroreceptor pore there is an electroreceptor cell whose changes when V,, IS altered by only a few m~crovolts[Adapted from
apical membrane has a low electrical resistance compared with that Bennett, 1968 ]
of the basal membrane. (C) Receptor cells release transmitter mole-
contact with axons of the eighth cranial nerve that inner- response to changes in V, of the receptor cell of as little as
vate the lateral-line system. The cell membrane facing the several microvolts.
exterior has a lower electrical resistance than does the basal The train of current pulses flows through the wa-
membrane, so most of the potential drop caused by the cur- ter from the posterior to the anterior end of the fish
rent moving across the cell occurs across the basal mem- (Figure 7-38).Any object whose conductivity differs from
brane, depolarizing it. Depolarization of the basal mem- that of water will distort the lines of current flow. The lat-
brane activates Ca2+channels in the membrane, and the eral-line electroreceptors detect the distribution of current
resulting influx of Ca2+at the base of the cell increases the flowing back into the fish through the lateral-line pores on
release of synaptic transmitter by the receptor cell. This the head and anterior end of the body and can detect the
transmitter increases the frequency of APs in the sensory changes in field produced by objects in the water. This sen-
fiber that innervates the receptor. Conversely, a current sory information is then processed in the greatly enlarged
flowing out of the body of the fish hyperpolarizes the basal cerebellum of the fish, enabling it to detect and locate ob-
membrane of the receptor cell and decreases the release of jects in its immediate environment.
transmitter below the spontaneous rate. Thus, the firing fre- Electrical signals are produced by other species of fishes
quency in the sensory fiber goes up or down, depending and used for quite a different task. In contrast with the
on the direction of the current flowing through the elec- weakly electric fishes that use electrical fields for navigation
troreceptor cell (see Figure 7-37B and C). The sensitivity of and signaling, some eels, torpedoes, and other fishes
these receptors and their sensory fibers, like that of the hair produce a powerful discharge of current to stun enemies
cells of the vertebrate ear, is truly remarkable. As seen in and prey. These strongly electric fishes produce a continu-
Figure 7-37C, changes in sensory nerve discharge occur in ous series of synchronous, relatively high frequency
250 PHYSIOLOGICAL PROCESSES
...............................................................................
Figure 7-38 Electroreception allows electric fishes to
recognize and to locate objects In their environment.
An object that has a conductivity greater than that of
water deflects current toward the axls of flow. An
object whose conductivity is lower than that of water
(inset) diverts the current away from the axis of flow.
[From "Electric Location by Fishes," by H. W. Lissman.
Copyright 0 1963 by Scientific American, Inc. All
rights resewed.]
Lateral-line system
depolarizations with their electric organs, and the way in tions, and the endings appear to detect changes in tissue
which the electrical discharges of these fishes are generated temperature, rather than the radiant energy itself. The
and used resembles the way in which muscles are con- mechanisms by which temperature changes can alter re-
trolled to produce movement (see Chapter 10). ceptor output are not known. The sensory axons from the
pit organs of the rattlesnake increase their firing rate tran-
siently when the temperature inside the pit increases as lit-
THERMORECEPTION
tle as 0.002"C, and this change in receptor firing rate can
Temperature is an important environmental variable, and modify behavior. For example, a rattlesnake can detect the
many organisms acquire sensory information about tem- radiant heat emitted by a mouse that is 40 cm away if the
perature from the action of specialized nerve endings, body temperature of the mouse is at least 10°C above the
or thermoreceptors, in the skin. Higher-order neurons re- ambient temperature. Furthermore, the temperature recep-
ceive input from thermoreceptors and contribute to the tors lie deep within the facial pits, and this arrangement al-
mechanisms that regulate the temperature of the body (see lows the snake to detect the direction of a source of radiant
Chapter 16).In addition, some of the neurons in the hypo- heat (Figure 7-39B).
thalamus of vertebrates are able to detect changes in body Both the external skin and the upper surface of the
- -
Temperature (OC)
1n6er ~rdnnchof
chamber trigeminal nerve
B Object in front
stimulates both pits
Time (s)
VISION
Figure 7-39 The facial pits of rattlesnakes contain extremely sensi- Since the Earth formed more than 5 billion years ago, sun-
tive thermoreceptors. (A) Structure of a facial pit in the rattlesnake
Crotalus viridis. (B) The position of the facial pits makes thermoreception
light has been an extremely potent selective force in the evo-
by the pit organs directionally sensitive. [Adapted from Bullock and lution of living organisms, and most organisms are able to
Diecke, 1956.1 respond to light in some way. Photoreception consists of
transducing photons of light into electrical signals that can
be interpreted by the nervous system, and photoreceptive
gories of thermoreceptors are distinguished from one an- organs-typically called eyes-have evolved into many
other in accord with how they respond to temperature shapes and sizes and with many distinct designs. Interest-
changes near the normal temperature of the human body ingly, although the physical structure of eyes varies greatly
(about 37°C). Both warmth and cold receptors increase among species, visual transduction is based upon a very
their firing rate as the temperature becomes increasingly dif- highly conserved set of protein molecules that provide an
ferent from 30"-35°C (Figure 7-40A): warmth receptors optical pathway leading light to the photoreceptive surface
fire faster as the temperature gets warmer; cold receptors and that capture photons within the photoreceptors.
fire faster as the temperature gets colder. However, when This conservation of visual molecules suggests that,
the temperature becomes sufficiently different from when suitable biochemical means had evolved to solve the
252 PHYSIOLOGICAL PROCESSES
............................................................................
problem of capturing light's energy, the sequences were mon evolutionary descent, although they differ to some ex-
conserved even though they were packaged into organs tent because the two species live in different optical media.
with highly diverse structural properties. For example, the The evolution of eyes has proceeded in two stages. Vir-
opsins are protein visual pigment molecules. Each molecule tually all major animal groups have evolved simple eyespots
includes seven transmembrane domains. Opsins are cou- consisting of a few receptors in an open cup of screening
pled to photopigment molecules, which are structurally al- pigment cells (Figure 7-41A). Some biologists estimate that
tered by the absorption of photons and which in turn mod- such photon detectors have evolved independently between
ify the properties of the opsin protein (see Figure 7-3). 40 and 65 times. Eyespots provide information about the
Opsins are found widely in the animal kingdom, even in surrounding distribution oflight and dark, but they do not
photoreceptive structures that are extremely simple and provide enough information to allow detection of either
that lack the features that would constitute an eye. In many predators or prey. For pattern recognition or for controlling
organisms, the structure of the eye has evolved to collect locomotion, animals need an eye with an optical system that
and focus incident light rays before they arrive at the site of can restrict the acceptance angle of individual receptors and
transduction. Eyes refract light through highly concen- form some kind of image. This stage of optical evolution
trated soluble proteins that are formed into lenses, and happened less frequently, occurring in only 6 of the 33 meta-
these refractive structures also have an interesting evolu- zoan phyla (Cnidaria, Mollusca, Annelida, Onychophora,
tionary history. We first consider how eyes collect and fo- Arthopoda, and Chordata). Because these phyla contain
cus light. about 96% of all extant species, it is tempting to speculate
that having eyes confers significant selective benefits.
Optic Mechanisms: Evolution and Function Ten optically distinct designs for image-formingeyes have
The physics of light tightly constrains the structure of an been discovered to date. They include nearly all the possibil-
eye that will produce a usable image. Most of the possible ities known from physical optics except Fresnel and zoom
designs have been "discovered" in the course of evolution, lenses. In addition, there are some variations, such as array
giving rise to similar structures in unrelated animals. One optics, that have not been used by physicists studying optics.
of the most well known examples of convergent evolution Simple eyespots are typically less than 100 pm in di-
is the similarity of eyes in the phylogenetically unrelated ameter and contain between 1 and 100 receptors. Even
squid and fishes. These eyes are similar in many details be- simple eyespots allow some visually guided behavior. In
cause optical laws have dictated convergent solutions to the protozoa and flatworms, the direction of a light source is
problem of seeing under water. In contrast, the eyes of hu- detected with the help of a screening pigment that casts a
man beings and fishes are similar because they share com- shadow on the photoreceptors. Some flagellates, for exam-
6
slmplest eye conslsts of a shallow open plt
llned wlth photoreceptor cells (B) In sllghtly
-
more complicated eyes, the aperture of the
eye IS small In proportion to the slze of the
eye, and the eye operates slmllarly to a pin-
+ hole camera (C)An alternative Improvement
allowlng Image format~onIS the addltlon of a
refracting element between the aperture and
Shallow pit the layer of photoreceptors (D)Three lenses
arranged In serles Improve the optlcal prop-
Vertebrate
ertles of the eye In Pontella, a cope-
eye
B
I' D
pod (E) The vertebrate eye IS an elaborat~on
of a slmple eye to wh~cha small aperture and
a lens have been added Arrows ~nd~cate pos-
slble evolutionary relations among types of
eyes [Adapted from Land and Fernald, 1992.1
Vertebrate eye
254 PHYSIOLOGICAL PROCESSES
.......................................
each unit in a compound eye is relatively large, compound tally accessible and its activity could be monitored with
eyes have lower visual acuity than do vertebrate eyes. How- simple electrical recording techniques.
ever, although the mosaic image formed by this type of The visual receptor cells of the Limulus compound
eye is coarser than the image produced by a vertebrate eye eye are located at the base of each ommatidium
(Figure 7-42B), it is certainly recognizable. (Figure 7-43B and C). Each ommatidium lies beneath a
hexagonal section of an outer transparent layer, the corneal
The eyes of Limulus lens. The primary photoreceptors are 12 retinular cells,
Each ommatidium of a compound eye contains several which surround the dendrite of another neuron, the eccen-
photoreceptors. The most intensively studied invertebrate tric cell. Each retinular cell has a rhabdomere, in which the
photoreceptors are those in the lateral eyes and the ven- surface membrane of the cell is thrown into a dense profu-
tral eye of the horseshoe crab Limulus polyphemus sion of rniaovilli, which are miniature tubular evaginations
(Figure 7-43). The two lateral eyes of Limulus are typical of the surface membrane (see Figure 7-43D). The microvilli
compound eyes, similar to those in Figure 7-42A, whereas greatly increase the surface area of the cell membrane in the
the unpaired ventral eye is simpler in structure and more rhabdomere. Light enters through the lens and is absorbed
like the eyespot shown in Figure 7-41A. Most of the early by molecules of the photopigment rhodopsin that are lo-
electrical recordings made from single visual units were cated in the receptor membrane within the rhabdomere.
done with this lateral eye, because the eye was experimen- Transient, random depolarizations of the membrane po-
C
Light
eccentric cell
- t
L~ght
on
1
Light
off
Retinular cell
Microvilli of
rhabdornere
Figure 7 4 4 The structure of ommatidia allows some arthropods to per- formed by two sets of microvilli.The upper microvilli were sectioned par-
ceive the plane of polarized light. (A)The interdigitating rhabdomeresof allel to their longitudinal axis, and those in the lower set were sectioned
separate retinular cells produce two sets of mutually perpendicular mi- perpendicular to their longitudinal axis. [Part A adapted from Horridge,
crovilli. (B) Electron micrograph of a section through the rhabdome 1968; part B courtesy of Waterman et al., 1969.1
256 PHYSIOLOGICAL PROCESSES
...............................................................................
SPOTLIGHT 7 - 1 judgments made by a human subject who IS asked t o com-
pare the lntensltles of different l ~ g hstlmuli
t
SUBJECTIVE CORRELATES 2. A receptor's response t o flashes of 11ghtthat are less than
1 second In duratlon IS proportional t o the total number of
OF PRIMARY photons In the flash, regardless of the actual duratlon That
the number of lmpulses generated remalns constant as
PHOTORESPONSES IS,
-
+
-c (A) When l~ghtflashes are shorter than 1
a
The Vertebrate Eye
Vertebrate eyes (see Figure 7-41E) have certain structural
features similar to those of a camera. In a camera, the image
Cell a is focused on the film by moving the lens forward or back-
ward along the optic axis. For example, to bring objects that
are close to the camera into focus, the lens must be posi-
cd b w- - tioned relatively far away from the film. To focus on distant
objects, the lens is moved forward. In the vertebrate eye, in-
I I I I 1 , I J cident light is focused in two stages. In the initial stage, in-
400 500 600 700
500 600 '
0
° cident light rays are bent as they pass through the clear outer
Figure 7-45 The response of crayflsh photoreceptors to polarized light surface of the eye, called the cornea (Figure 7-46). They are
varies with the plane of polarization. Two cells, a and b, were presented
with a series of equal-energy flashes of polarized light at various wave-
further bent. or refmcted, as they, Dass through a second-
structure, the lens, and finally form an inverted image on the
lengths. The color of light in each flash (indicated by its wavelength in
nanometers) is indicated along the lower axis. Cell a responded maxi-
rear internal surface of the eye, the retina. In fact, most of
rnally to light with a wavelength of about 600nrn; cell b responded max- the that Occurs in the eye 85% the
imally to light of 450 nm. When the plane of ~olarization(red arrow) was tal) takes place at the air-cornea interface, and the rest de-
perpendicular to the microvilli, the responses in both cells were small pends on the effect of the lens. Like a camera, certain bony
(left).Responses of both cells were enhanced when the plane of polar- fishes focus images on the retina by moving the lens of the
ization (red arrow) was rotated so as to lie parallel to t6e microvilli (right).
[Adapted from Waterman and Fernandez, 1970.1
eye with respect to the retina. (This principle of changing the
distance between the lens and the light-receptive surface has
also been adapted by some invertebrates. For example, in
were oriented systematically in the microvilli and each pref- the eyes of jumping spiders, the position of the lens is fixed,
erentially absorbed light with its electric vector parallel to and focusing depends on moving the retina.)
the microvilli, the anatomical arrangement within the rhab- In contrast, neither the lens nor the retina can be moved
dome could provide a physical basis for the detection of in the eyes of higher vertebrates. Rather, the image is
Rectus
tendon
Figure 7-46 In the mammalian eye, incident light is refracted by the by the lens. The lens is held in place by the zonularfibers. When ciliary
cornea and the lens and is focused on the photosensitiveretina. In this di- muscle fibers contract, tension on the zonular fibers is reduced, and the
agram, the refraction of light has been simplified; refraction at the air- elastic properties of the lens cause it to become more rounded, short-
cornea interface is omitted. The image focused on the retina is inverted ening the focal length.
258 PHYSIOLOGICAL PROCESSES
...............................................................................
focused by changing the curvature and thickness of the ter opens. The vertebrate eye has an opaque iris with a vari-
lens. Changing the curvature of the lens surfaces changes able aperture called the pupil, which is analogous to the
the distance at which an image passed through the lens mechanical diaphragm of a camera. When circular smooth
comes into focus, called the focal length of the lens. The muscle fibers in the iris contract, the pupillary diameter de-
shape of the lens is changed by modification of the tension creases, and the proportion of incident light that is allowed
exerted on the perimeter of the lens. The lens is held in to enter the eye is reduced. Contraction of radially oriented
place within the eye by the radially oriented fibers of the muscle fibers enlarges the pupil. The contraction of these
zonula (seeFigure 7-46).The fibers of the zonula exert out- muscles-and, hence, the diameter of the pupil-is con-
wardly directed tension around the perimeter of the lens. trolled by a central neuronal reflex that originates in the
Radially arranged ciliary muscles adjust the amount of ten- retina. This pupillary reflex can be demonstrated in a dimly
sion exerted on the lens. When the ciliary muscles relax, the lit room by suddenly illuminating a subject's eye with a
lens is flattened by elastic tension exerted by the fibers of the flashlight.
zonula, which pull the perimeter of the lens outward. In this Changes in pupillary diameter are transient. After a re-
state, objects far from the eye are focused on the retina, but sponse to a sudden change in illumination, the pupil grad-
objects close by would be fuzzy. Objects close to the eye are ually returns after several minutes to its average size. More-
brought into focus on the retina when the ciliary muscles over, the area of the pupil can change only about fivefold,
contract, relieving some of the tension on the lens and al- making it no match for the changes in the intensity of illu-
lowing the lens to become more rounded. This process is mination normally encountered by the eye, which equal six
called accommodation to close objects. The ability to ac- or more orders of magnitude. Thus, although the pupil can
commodate decreases with age in human beings as the lens produce rapid adjustments to moderate changes in light in-
becomes less elastic, producing a type of "far-sightedness," tensity, other mechanisms must be available. The eye
called presbyopia. adapts to extremes of illumination by changes in the state
Perhaps the most remarkable thing about accommo- of visual pigments and by the processes of neuronal adap-
dation is not the mechanical mechanisms for altering the tation (see Mechanisms of Adaptation earlier in this chap-
focal length of the lens, but the neuronal mechanisms by ter). Pupillary constriction provides an additional advan-
which a "selected" image-out of all the complexity in the tage: the quality of the image on the retina improves. The
visual environment-is correctly focused on the retina as a edges of the lens are optically less perfect than is the center;
result of nerve impulses to the ciliary muscles. A related so, when the pupil is constricted, light is prevented from
neuronal mechanism produces binocular convergence, in passing through the perimeter of the lens and optical aber-
which the left and right eyes are positioned by the ocular rations are reduced. The depth of focus (the distances
muscles in such a way that the images received by the two through which objects are in focus when the lens is in one
eyes fall on analogous parts of the two retinas, regardless fixed shape) increases with decreased pupillary diameter,
of the distance between the object and the two eyes. When just as it does in a camera when the aperture is reduced.
an object is close, each of the two eyes must rotate toward
the middle of the nose; when an object is far away, the two Visual receptor cells of vertebrates
eyes rotate outward from the midline. The stimulus to all visual receptor cells is electromag~ietic
radiation that falls within a particular range of energy,
Responses to changes in light intensity called visible light (Figure 7-47). The energy of electro-
In a camera, the intensity of light admitted to the film is magnetic radiation varies inversely with its wavelength, and
controlled by adjusting the aperture of a mechanical di- we perceive this variation in energy as variation in color. Vi-
aphragm through which light is admitted when the shut- olet light, the highest energy of light to which the human
Log of wavelength
Pigment epithelium
cilium
Figure 7-48 Vertebrate photoreceptors are classified as rods or cones from the source of light. The pigmentthat absorbs the light energy is con-
on the basis of their morphological and physiological properties. The tained in membrane lamellae, and the ends of the outer segments lie
outer segments of rods and cones, where light is captured, face away against the pigment epithelium.
260 P H Y S I O L O G I C A L PROCESSES
...............................................................................
information about the visual world, a characteristic called cones of mammals and some other vertebrates, the lumen
high visual acuity. In human beings and in certain other of each lamella is open to the cell exterior. In rods, the
mammals, the fovea contains only cones, whereas the lamellae pinch off completely from the surface membrane
remainder of the retina contains a mixture of rods and of the outer segment so as to form flattened sacs, or disks,
cones, with the rods significantly outnumbering the cones. that are stacked like pita bread on top of one another
In mammals, cones mediate color vision, and rods, which within the rod outer segment. The stack of disks is com-
are more light-sensitive, mediate only achromatic vision. pletely contained within the surface membrane of the visual
This distinction between rods and cones does not, however, cell. The photopigment molecules are embedded in the.
pertain to all vertebrates. In fact, the retina in some species membranes of the disks. Because the photopigment lies in
contains only rods, based on morphology, but may the disk membranes of the rod outer segment but not in the
nonetheless be capable of color vision. surface membrane, the primary step in photochemical
Rods and cones are structurally and functionally more transduction must take place in the d s k membranes, rather
similar to one another than are the wide variety of pho- than in the surface membrane.
toreceptors found in the invertebrates. Each vertebrate The eyes of many invertebrateslack the ciliary structure
receptor cell contains a segment with an internal struc- that connects the inner and outer segments of vertebrate
ture similar to that of a cilium. This rudimentary cilium rods and cones (Figure 7-49).In these invertebrate eyes, the
connects the outer segment, which contains the photore- photopigment is located in microvilli formed by the cell
ceptive membranes, to the inner segment, which contains membrane, and these pigment-containing microvilli make
the nucleus, mitochondria, synaptic contacts, and so forth up the rhabdomeres. Because many invertebrate species
(see Figure 7-48).The receptor membranes of vertebrate vi- have simple eyes in which the photoreceptors are of the
sual cells consist of flattened lamellae derived from the sur- rhabdomeric type, it might be tempting to conclude that
face membrane near the origin of the outer segment. In the rhabdomeric photoreceptors are found only in simple eyes.
Rhabdomeric line
However, the eyes of the octopus are optically very com- it is maximal in the dark, are kept from accumulating in
plex and the photoreceptors are rhabdomeric. In addition, the cell by the steady action of the metabolically driven
some bivalve mollusks (e.g., the scallop, Pecten, and the Na+,K+ATPase. A dark current is found only in vertebrate
clam, Lima) have eyes with two separate layers of pho- photoreceptor cells and not in invertebrate photoreceptors.
toreceptors. One layer contains ciliary receptors, and the After light absorption by the photopigment, the con-
other contains rhabdomeric receptors. ductance to sodium, gNa,of the outer segment decreases,
causing the dark current to decrease and V,,, to hyperpo-
larize toward EK(see Figures 7-50B and 7-51B). When the
light stimulus stops, gNaof the membrane returns to its high
resting level, and V,,, becomes more positive, returning to
its resting level between ENaand EK.The change in V,,, at
the onset of light is carried electrotonically (see Passive
Spread of Electrical Signals in Chapter 6) into the inner seg-
ment of the photoreceptor. In the inner segment, changes in
V,, modulate the steady release of neurotransmitter from
In all photoreceptor cells, the transduction of light en- the presynaptic sites located in the basal part of the inner
ergy produces a change in the membrane potential; how- segment. Like vertebrate auditory receptors, vertebrate
ever, the effect of the transduction is different in vertebrate photoreceptors lack axons. They synapse onto other neu-
and invertebrate photoreceptors. Invertebrate photorecep- rons, which carry the visual signal toward the central ner-
tors depolarize in response to light (Figure 7-50A; see also vous system. The neuronal signal is passed along by other
Figure 7-45), but vertebrate rods and cones hyperpolarize neuronal cells of the retina, ultimately influencing the ac-
in response to a light stimulus (Figure 7-50B). Membrane tivity of axons that project to the brain within the optic
conductance measurements before and during illumination nerve. It is interesting that a hyperpolarization, rather than
have shown that the effect of light on vertebrate photore- a depolarization, is produced when a vertebrate photo-
ceptors is to decrease the conductance for sodium, gNa,of receptor absorbs light, because in most sensory systems re-
the outer segment membrane. In the dark, the surface mem- ception of a stimulus depolarizes the receptor cell. In verte-
brane of the vertebrate rod outer segment is nearly equally brate photoreceptors, the inner segment steadily secretes a
permeable to Na+ and K+, and V, lies about halfway be- transmitter while it is being partially depolarized by the
tween EKand EN,. In this state, Na+ ions leak into the dark current. The hyperpolarizationthat occurs in response
outer segment through channels that are steadily open in to illumination decreases the amount of transmitter re-
the dark (Figure 7-51A). The Na+ ions that carry this leased onto the next neuron in line, modifying the activity
inward current, which is called the dark current because of that second-order neuron.
The change in membrane potential that is produced
in a group of photoreceptors when they are illuminated
can be recorded by extracellular electrodes, as can action
A Invertebrate B Vertebrate potentials traveling down the axons of a nerve. Many
photoreceptors are tiny cells making intracellular recording
difficult, so this method of recording-called an electro-
retinogram-has been extremely useful in the study of vi-
sion (Spotlight 7-2).
nn Light o n Light on
ried to the central nervous system. The process of visual
transduction has received an enormous amount of research
attention, and the features of the visual process have pro-
Figure 7-50 Most invertebrate photoreceptors depolarize in re- vided clues to physiologists studying sensory transduction
sponse to a stimulus, whereas vertebrate photoreceptors hyperpolarize. in other sensory modalities. Studies of photoreception have
(A) Transduction of light energy into chemical energy w ~ t h ~most
n inver- been carried out in many different species spanning several
tebrate photoreceptors causes an increase in the permeability of the sur-
phyla. Many similarities between vertebrate and inverte-
face membrane to Na+ and K+, depolarizing the cell. (B)Vertebrate pho-
toreceptors respond to light with a decrease in gNaof the surface
brate photoreception have been found, although it is now
membrane, leaving a residual low g,and shifting V, toward EK.As a re- thought that invertebrate photoreception may be more
sult, the cell hyperpolarizes. complex because it relies on two light-activated pathways,
Figure 7-51 Illumination reduces the dark current in ver-
tebrate rods. The g,, of the rod outer segment is high in
the dark(A) and becomes reduced in the light (B). Forthis
reason, the dark current, which is carried by Na+ ions that
In 4 Out leak into the outer segment, drops during illumination. In
the equivalent circuit (top left), the battery is the Na+, K+
ATPase, and the light-activated variable resistor (R,,) rep-
resents the gNaof the outer segment. [Adapted from
Hag~ns,1972.1
N a conductance
Outer reduced
segment +----
4
I
'--I I I I
4 I
I
1
I
I I
Inner I I
segment I I Dark
Nucleus -
-, -- -,I
I
Synaptic
ending
A Dark 6 Light
rather than the single pathway found in vertebrates. There with wavelengths between cm and lo-' cm is termed
are other, perhaps related, differences as well. For example, light. Only a small part of this segment of the spectrum-
capture of a single photon by a photoreceptor in Limulus ranging from about 400 nm to about 740 nm-is visible to
produces a peak current of -1 nA, whereas capture of a human beings. Below this range is the ultraviolet (W)part
single photon by a vertebrate rod photoreceptor changes of the spectrum, and above it the infrared (IR),neither of
the current by -1 pA, three orders of magnitude smaller. which is visible to human beings and other mammals.
Moreover, invertebrate photoreceptors may respond to There is nothing qualitatively special about those parts
light intensities spanning seven orders of magnitude, of the spectrum that renders them invisible to us. Rather,
whereas vertebrate rods respond to intensities only within what we see depends on which wavelengths are absorbed
four orders of magnitude. Despite these differences in de- by our visual pigments. For example, there is a condition
tail, all types of photoreceptors have been shaped by evo- called cataract, in which the lens becomes opaque. Treat-
lution to convert the energy of photons into neuronal en- ment of the condition consists of surgically removing the
ergy, and studies of all types of eyes have contributed to our lens; after this surgery, patients can see light into the W
understanding of the process. range because it is absorption of UV light by the lens that
prevents people from seeing those wavelengths. The com-
Visual pigments pound eyes of many insects can detect light into the UV
The spectrum of electromagnetic radiation extends from range, causing some flowers containing W-reflecting pig-
gamma rays, with wavelengths as short as 10-l2 cm, to ments to look much less plain to insects than they do to
radio waves, with wavelengths greater than lo6 cm (see mammals, but all animals are sensitive to only a part of the
Figure 7-47). The segment of the electromagnetic spectrum spectrum of electromagnetic radiation that is available in
S P O T L I G H T 7-2 activity of the photoreceptor cells and other neurons in the
retina. It took several years t o sort out the source of each of the
THE components of the ERG, but we now believe that the a wave is
due t o the receptor current produced by the visual receptor
ELECTRORETINOGRAM cells. The b wave follows the a wave, and it is produced by elec-
trical activity of the second-order retinal neurons that receive in-
In a teaching laboratory, it is sometimes useful t o record the p u t from the receptor cells. The c wave is found only in verte-
summed electrical activity of the eye, which is technically much brates and appears t o b e produced by the pigment epithelial
less complicated than recording from single cells with micro- cells against which the outer segments of the visual cells abut. In
electrodes. The recording electrode (which can be a thread or a the developing eyes of tadpoles, the ERG consists of only an a
wick that is saturated with saline) is placed on the cornea, and the wave before synaptic contacts are established. Similarly, in the
ground electrode is attached t o another part of the body. When eye of an adult frog, if synaptic transmission between the photo-
a light is flashed on the eye, a complex waveform is recorded by receptors and the second-order neurons is blocked pharmaco-
the electrode (as shown in the adjoining figure). This recording is logically, the ERG consists of only an a wave.
called an electroretinogram (ERG), and it records the summed
1 Off
recording [Adapted from Brown, 1974 ]
sunlight. Perhaps the visual pigments of vertebrates absorb same process that is used in the photosynthetic conversion
only a limited range of the electromagnetic spectrum of of radiant energy into chemical energy in plants.
sunlight because vertebrate life evolved in water, which
heavily filters electromagnetic radiation. The range of the Photochemistry of visual pigments
spectrum to which vertebrate photopigments-including The energy content of visible light is just low enough to be
those of terrestrial mammals such as human beings-are absorbed by molecules without breaking them up. The con-
sensitive matches rather closely the spectrum of light that is cept that a pigment is essential for the process of absorbing
admitted through water. light and transducing its electromagnetic energy into chem-
All known organic pigments owe their ability to absorb ical energy originated with John W. Draper, who concluded
light selectively to the presence of a carbon chain, or ring, in 1872 that, to be detected, light must be absorbed by mol-
that contains alternating single and double bonds. When a ecules in the visual system. R. Boll found soon thereafter
photon is captured by one of these molecules, the energy that the characteristic reddish purple color of the frog's
state of the molecule is changed. The energy contained in retina fades (bleaches)when the retina is exposed to light.
a quantum of radiation is equal to Planck's constant di- The light-sensitivesubstance, rhodopsin, that is responsible
,vided by the wavelength, A (in centimeters): for the purple color, was extracted in 1878 by W. Kiihne,
who also found that, after the pigment has been bleached by
light, its reddish purple color can be restored by keeping the
retina in the dark, provided that the receptor cells remain in
Thus, the energy in a photon increases as the wavelength of contact with the pigment epithelium at the back of the eye.
radiation decreases. Quanta with wavelengths less than Since then, much has been learned about the chemical
1 nm contain so much energy that they break chemical nature and physiological effects of rhodopsin. It absorbs
bonds or even atomic nuclei; quanta with wavelengths light maximally at wavelengths of about 500 nm. It is
greater than 1000 nm lack sufficient energy to affect mo- found in the outer segments of rods in many vertebrate
lecular structure. The visual pigments absorb maximally species and in the photoreceptors of many invertebrates.
between these two limits. When a quantum of radiation is Rhodopsin molecules are packed at high density into re-
absorbed by a photopigment molecule, it raises the energy ceptor membranes; there may be as many as 5 x 1012mol-
state of the molecule by increasing the orbital diameter of ecules per square centimeter, which is equivalent to an in-
the electrons associated with a conjugated double bond, the termolecular spacing of about 5 nm.
All known visual pigments consist of two major com- When light hits the photopigment, an intermediate,
ponents: a protein (opsin) and a light-absorbing molecule. metarhodopsin II, is formed. Metarhodopsin I1 activates
In all instances, the light-absorbing molecule is either reti- another protein that is associated with the membrane and
nal or 3-dehydroretinal (Figure 7-52). Retinal is the alde- that binds GTP in exchange for GDP. This protein, which
hyde of vitamin A,, a carotenoid. Vitamin Al is an alcohol we now know belongs to the family of G proteins, is called
and is also called retinol; 3-dehydroretinal is the aldehyde transducin in recognition of its key role in the transduction
of vitamin A,, which is also called 3-dehydroretinol. In ad- of light. The activated subunit of transducin diffuses in the
dition to its major components, rhodopsin includes a six- plane of the membrane, encountering many phosphodi-
sugar polysaccharide chain and a variable number (as esterase molecules, which hydrolyze cGMP to 5'-GMP. In
many as 30 or more) of phospholipid molecules. The lipo- vertebrate photoreceptors, the dark current Na+ channels
protein opsin, which binds the phospholipids and the poly- are open only in the presence of cGMP; so, when cGMP is
saccharide chain, appears to be an integral part of the pho- hydrolyzed, these channels close (Figure 7-53). The mem-
toreceptor membrane. Carotenoid molecules move back brane of the rod outer segment contains a class of channels
and forth between the photoreceptor membrane and the that are permeable to three cations: Na+, Mg2+and Ca2+.
pigment epithelium at the back of the retina during bleach- When the level of cGMP drops, the conductance through
ing and regeneration of the visual pigment. (Incidentally, these channels drops. Most importantly, the inward I,=
the pigment that confers a dark color on the pigment ep- drops, and the residual K+ current through other channels
ithelium is photochemicallyinactive and is unrelated to the causes the cell to hyperpolarize. When the light stimulus
visual pigment. Instead, it keeps light from scattering and ends, cGMP is regenerated by the action of another en-
reflecting diffusely back toward the retina.) zyme, guanylate cyclase. As the level of cGMP rises, the
The retinal molecule assumes two sterically distinct dark current channels open, and the receptor current re-
states in the retina. In the absence of light, the opsin and the turns to its full value in the dark. Activated transducin col-
retinal are linked covalently by a Schiff's base bond, and lides with, and activates, phosphodiesterase molecules at a
retinal is in the 11-cisconfiguration (seeFigure 7-3). When rate of about lo6 molecules per second, allowing the cap-
the 11-cis retinal captures a photon, it isomerizes into the ture of a single photon to affect the conductance through
all-trans configuration (see Figure 7-52). This cis-trans an enormous number of ion channels. This numerical rela-
isomerization is light's only direct effect on the visual pig- tion generates an impressive amplification between the cap-
ment. The conversion from 11-cis to all-trans retinal initi- ture of a single photon and the effect that the event pro-
ates a series of changes in the relation between the retinal duces on V,.
and the opsin protein, including changes in the conforma- After the cis-trans isomerization of retinal has oc-
tion of the opsin itself. curred, further changes in the molecule appear to be irrel-
l H l H
HzC
/c\ C
/C\
C
/C\c/$\cH
I I H H I
H2C\ /C\
c I CH, H,C/'\CH
CH, I
HC
\o
Figure 7-52 The carotenoid pigment retinal changes its steric confor- space-f~ll~ng
and llne d~agramsof the molecular structure are shown [Part
mation when it absorbs a photon. (A) In the dark, the bonds of carbon 11 B from "Molecular Isomers In Vls~on,"by R Hubbard and A Kropf Copy-
are arranged in the cis configuration. (B) When a photon is captured, r~ght0 1967 by Scient~flcAmer~can,Inc All r~ghtsresewed.]
these bonds are converted into the straight, all-trans configuration. Both
SENSING T H E ENVIRONMENT 265
............................
Figure 7-53 When light is absorbed by
Light retinal, a series of reactions causes the Na+
channels that carry the dark current to close.
(A) Activated rhodopsin increases the activity
of a G protein, transducin. The activated G
protein then activates many phosphodi-
esterase (PDE) moiecules, reducing the intra-
cellular concentration of cyclic guanosine
monophosphate (cGMP),which leads to the
closing of Na+ channels that carry the dark
current. The receptor cell then hyperpolarizes.
(B) Recordings of currents in single rod pho-
Dark current toreceptors isolatedfrom toad retina. (Left) A
-
5'-GMP Na+ channel flash of light causes the inward dark current of
10 pA to drop to zero (R~ght) The photore-
ceptor has been broken open, and the exter-
B nal sal~nehas been changed to rnlrnlc Intra-
-
0.8 mM cGMP cellular lonlc concentrat~onsWhen cGMP IS
added to the external sallne (exposing the In-
-
side of the outer segment to a hlgh concen-
& -10 tratlon of cGMP), a very large Inward current
t C
develops [Part B adapted from Yau and
E
L
g -200
L
Flash Nakatan~,1985.1
5 5 '
0 5 10 15 0 20 40
Time (s) Time (s)
evant to the excitation of visual receptor cells, but the periments, rods are teased apart and one of them is drawn
subsequent reactions (Figure 7-54)are necessary for regen- into a recording pipette where it is stimulated by a small
erating active rhodopsin. Activated rhodopsin is hy- beam of light. When the stimulating light is very dim, it is
drolyzed spontaneously to retinal and opsin, which are possible to record small current fluctuations, each of which
both reused repeatedly. Free retinal is reisomerized back occurs when a single rhodopsin molecule has been photo-
into the 11-cis form and reassembled with an opsin to form isomerized by a single photon. The properties of the current
rhodopsin. Any retinal that is lost or chemically degraded recorded under these conditions are similar to the proper-
in the process is replenished from vitamin A, (retinol) ties of the current measured through a single acetylcholine
stored in the cells of the pigment epithelium, which actively receptor channel at the neuromuscular junction. (The
take up the vitamin from the blood. A nutritional deficiency change in current that is associated with the capture of one
of vitamin A, decreases the amount of retinal that can photon is about 1 PA.) Because photoreceptors can re-
be synthesized and, hence, decreases the amount of avail- spond to a single photon, or quantum of energy, the sensi-
able rhodopsin. The result is reduced photosensitivity of tivity of photoreception is limited by the quanta1 nature of
the eyes, a condition that is commonly known as night light; there is no smaller amount of light than one photon.
blindness. Elucidating the process of visual transduction has
8 Rod photoreceptors can respond to the absorption of demonstrated the power of a comparative approach. Al-
a single photon, partly because rhodopsin is so densely though vertebrate and invertebrate photoreceptors seem
packed into their disc membranes. There are about 20,000 quite different from each other at the electrophysiological
rhodopsin molecules per square micrometer in the rod level, there are many similarities between them at the mo-
outer segment, which is much closer packing than, for ex- lecular level. The molecular genetics available in Droso-
ample, the density of acetylcholine receptors at the neuro- phila and the physiological accessibility of the vertebrate
muscular junction. By recording from a single rod, Denis retina have been combined to provide an array of enor-
Baylor, of Stanford University, has measured the response mously powerful experimental approaches to the question
to the capture of a single photon (Figure 7-55).In these ex- of how visual information is acquired and processed by
Il-cis , I
I
I
Il-cis
Retinol (vitamin A) is stored in the pigment
eplthellum and can be deltveredto the photo
Retinal I Retinol
I receptors for generating new rhodopsln
Retina ! Pigment epithelium molecules.
760 photons. pm-Z.s-'
I
light
'outer
segment
of rod
Time (s)
Figure 7-55 Rods can respond electrically to the capture of a single company the capture of single photons. As the light intensity is increased,
photon. (A) A single rod outer segment is sucked into a smooth glass (intensity is indicated above each recording), the responses become
pipette electrode and is illuminated by a narrow bar of light while the larger and smoother. The duration of the illumination is indicated by the
ionic current across the membrane is recorded by the pipette. (B) The bar under each recording. Membrane currents are in PA. [Adapted from
recorded membrane current changes in response to illumination. In very Baylor et al., 1979.1
dim light (bottom record), small individual changes in the current ac-
photoreceptors. If the molecular identities of the players action spectrum of a photoreceptor depends upon the ab-
had not been so strongly conserved through phylogeny, sorption properties of its visual pigment. In addition, results
teasing out the details of visual transduction would prob- of such experiments confirm that each photoreceptor syn-
ably have taken much longer. thesizes only one of the visual pigments. Light that con-
tains different wavelengths generates photochemical reac-
Cones and rods tions in a particular photoreceptor cell in proportion to the
The ability to distinguish color, rather than just perceiving amount of each wavelength absorbed; thus, a photorecep-
the visual world in shades of gray, is correlated with the tor cell is excited by different wavelengths in proportion to
possession of multiple visual pigments, each of which ab- the efficiency with which its pigment absorbs each wave-
sorbs maximally at a different wavelength (Spotlight 7-3). length. Any photon that is not absorbed can have no effect
In vertebrate species that have color vision, it has been on the pigment molecule; any photon that is absorbed
found that different groups of photoreceptors contain spec- transfers part of its energy to the molecule as described
trally identifiable visual pigments, and each class of pho- in Photochemistry of Visual Pigments in this chapter. Thus,
toreceptor has a distinctive action spectrum. That is, the it is possible to restate Young's trichromacy theory (see
electrical response of each photoreceptor, when it is illumi- Spotlight 7-3) in relation to cone photoreceptors and their
nated, is maximal at a particular wavelength and falls off photopigments: there are in the human retina three classes
when the wavelength of incident light is either raised or of cones, each of which contains one visual pigment that is
lowered. In many species for which action spectra have maximally sensitive to blue, to green, or to orange light.
been recorded, three classes of photoreceptors have been The electrical output of each class depends on the number
found. The action spectra for some species have then been of quanta that are absorbed by the pigment and can thus
compared with the absorption spectra of individual photo- contribute to the events of transduction. The sensation of
receptors. Absorption spectra for single photoreceptors color arises when higher-order neurons integrate signals re-
have been measured by a process, called microspectropho- ceived from the three classes of cones.
tometry, in which a tiny beam of light is focused on one Knowledge about the molecular basis of color recep-
photoreceptor at a time and the absorption properties of tion has grown enormously since 1984, when Jeremy
that cell are determined. Photoreceptors studied in this way Nathans described the molecular structure of human
fall into distinct classes for each species; there are no inter- opsins and thus provided an explanation for hereditary
mediate spectra, which implies that each photoreceptor color blindness. For example, point mutations in individ-
synthesizes a single visual pigment (Figure 7-56). Both the ual pigment genes cause defects in sensitivity to particular
action spectra and absorption spectra of photoreceptors wavelengths. Indeed, the molecular basis for differential
have been determined in many species, and the two kinds spectral sensitivity among the opsins has been characterized
of spectra match one another closely, confirming that the by using naturally occurring variants in visual pigments.
SENSING THE ENVIRONMENT 267
....................................
A 455 530 625
, (Blue) (Green) (Red)
Wavelength (nm)
Figure 7-56 Each class of cones in the carp retina has a distinctive action
spectrum. (A) Absorption spectra of individual cones in the retina of a
goldfish indicate that there are three separate visual pigments, each with
a distinctive absorbance peak. These measurements were made by mi-
crospectrophotometry,which allows the absorption spectrum of a single
photoreceptorto be measured. In human beings, the class of cones that
is equivalent to the red-absorbingcone in the goldfish absorbs maximally
closer to 560 nm, which is in the yellow part of the spectrum. (B) Electri-
cal responses of three single cones in the retina of a carp to flashes of d ~ f -
ferent wavelengths, as shown by the scale at the top. The wavelength that
produces a maximal response is different for each of the three cones.
(C)When the amplitude of activity in each cell shown in part B was plot-
ted as a function of wavelength, three classes of cones were revealed,
each with an action spectrum approximating one of the absorption spec-
tra in part A. [Part A adapted from Marks, 1965; parts B and C adapted
from Tomita et al., 1967.1
It appears that 11-cis-retinal(or 11-cis-3-dehydroreti- or a defect in, one of the cone opsin genes. With the use of
nal) is the light-absorbing molecule in all visual pigments, these molecular markers in conjunction with visual tests, it
and this prosthetic group is combined with different opsin is now possible to define the molecular basis of this per-
molecules to produce visual pigments with different ab- ceptual problem. For example, the high incidence of
sorption maxima. Differences in the amino acid composi-
tion of opsins-rather than variation in the light-absorb-
ing prosthetic group-produce rhodopsins with different
absorption maxima. Nathans and his co-workers discov-
ered three genes that encode for the opsins in human cones. Cvtosol
The gene encoding the protein part of the blue-absorbing
pigment is located on an autosomal chromosome, whereas
the two genes for the "red"-absorbing and green-absorbing
proteins are closely linked on the X chromosome. The Inside disk
"red" and green opsins differ at only 15 of 348 amino
acids, and each shares about half its amino acids with
rhodopsin in rods (Figure 7-57).On the basis of sequence
similarity, we can surmise that the genes for these pigments
probably arose from a common ancestral gene that under-
Figure 7-57 The two opsin proteins in the human rhodopsins that ab-
went duplication and divergence. A comparison of the
sorb maximally in the red and the green parts of the visible spectrum dif-
amino acid sequences suggests that, of the cone pigments, fer by only 15 amino acids, most of which are thought to be in membrane-
the blue-sensitive pigment arose first, followed by the red spannlng helices. In this diagram, these variable amino acids are red.
and the green. Color blindness is caused by an absence of, [Adapted from Nathans et al., 19861
SPOTLIGHT 7 - 3 ceptors would be stimulated maximally by separate monochro-
matic "red" and "yellow" wavelengths, respectively, and both
LIGHT, PAINT, AND would be stimulated to a lesser degree by monochromatic or-
ange light. Young proposed, in other words, that the sensation
COLOR VISION for "orange" results from the simultaneous excitation of "red"
receptors and "yellow" receptors. Young had no notion about
In 1666, Sir Isaac Newton demonstrated that white light is
the physiology of photoreceptors, making his insight truly
separated into a number of colors when it is passed through a
remarkable.
prism. Each spectral color is monochromatic; that is, it cannot be
Extensive psychophysical investigations carried out in
separated into yet more colors. At that time, however, it was al-
the nineteenth century by James Maxwell and Hermann von
ready known that a painter could match any spectral color (e.g.,
Helmholtz supported Young's trichromacy theory, and ad-
orange) by mixing two pure pigments (e.g., red and yellow), each
ditional support came from later investigations by William
of which reflects a wavelength different from that of the color
Rushton. However, direct evidence for the existence of three
produced. Thus, there seemed to be a paradox between
classes of color-receiving photoreceptors was missing. Then, in
Newton's demonstration that there is an infinite number of col-
1965, W. B. Marks and E. MacNichol measured the color ab-
ors In light and the growing awareness of Renaissance painters
sorption of single cone photoreceptors in the goldfish retina (see
that all colors could be produced by combinations of three pri-
Figure 7-56A). They found three classes of cones, each of which
mary pigments-red, yellow, and blue. This paradox appeared
absorbed maximally at a unique wavelength. Subsequent mea-
to be resolved by Thomas Young's suggestion, in 1802, that the
surements of the absorption spectra of cones in retinas from hu-
receptors in the eye are selective for the three primary colors:
man beings, monkeys, and other species of fishes reproduced
red, yellow, and blue. Young reconciled the infinite variety of
these results. It thus appears that the retinas of species that can
spectral colors that can be duplicated with the small number of
percelve and respond to color contaln photoreceptor cells w ~ t h
painter's pigments required to produce all colors by proposing
d~fferentabsorption spectra and that, In many of these species,
that each class of color receptor is excited to a greater or lesser
there are three d~sttnctclasses of receptors.
degree by any wavelength of light: The "red" and "yellow" re-
red-green color blindness arises from the recombination of image fall on the fovea, the star will fade or even disappear.
these closely linked red and green opsin genes. This increased sensitivity comes at a price: the broader con-
Color vision has been demonstrated in some members nections among rods reduces the acuity of rod-based vision.
of all classes of vertebrates. In general, retinas that include Our visual sensitivity is greatest when an image is focused
cone photoreceptors are associated with color vision, but onto the rods outside the fovea; our visual acuity is great-
examples of different color classes among rods have been est when we focus an image onto the cones of the fovea.
found. For example, frogs have two kinds of rods-red When visual pigments are explored throughout phy-
(containing rhodopsin, which absorbs in the blue green) logeny, some interesting patterns emerge. For example, all
and green (containing a pigment that absorbs in the visual pigments for which retinal is the prosthetic group are
blue)-in addition to cones. called rhodopsins. All human visual pigments-the rod
If color vision is mediated primarily by the cones, what pigment and the three cone pigments-are rhodopsins. Vi-
do rods contribute? Rods are more sensitive to light than sual pigments in which 3-dehydroretinal is the prosthetic
are cones (the records in Figure 7-55 were made from rods), group are called porphyropsins, and the distribution of the
and their connections to the next neurons in line are char- rhodopsins and porphyropsins among species shows an
acterized by greater convergence than are the connections interesting correlation with environment. All visual pig-
of cones (see Visual Processing in the Vertebrate Retina, ments of terrestrial vertebrates are rhodopsins. In addition,
Chapter 1I),producing greater summation of weak stim- rhodopsins are found among invertebrates, including
uli. Thus, rods are most effective in dim light. Because the Limulus, insects, and crustaceans. In contrast, porphy-
cones produce color vision, when only our rods are stimu- ropsins are found in the retinas of freshwater fishes, eury-
lated by dim light, we see in shades of black and gray, haline fishes (see Chapter 14),and some amphibians. This
rather than in color. In the human retina, most images are distribution suggests that some feature of the porphy-
prefentially focused onto the fovea, which contains only ropsins makes them particularly well adapted to conditions
tightly packed cones. Rods are found only outside the found in freshwater. In fact, anadromous fishes, which mi-
fovea. The differential distribution of rods and cones in the grate from freshwater to saltwater-or vice versa-during
retina causes us to be most sensitive to dim light when an their life cycle, change their visual pigment between por-
image is focused outside the fovea, onto parts of the retina phyropsin and rhodopsin during the migration. They syn-
where the rod population is higher. For example, a dim star thesize porphyropsin during their stay in freshwater and
will appear brighter if you adjust your gaze to make its im- rhodopsin while they are in saltwater. The absorption max-
age fall outside the fovea. If you s M your gaze to make the ima of the porphyropsins is shlfted toward the longer-
wavelength, red end of the visual spectrum, whereas part of the electromagnetic spectrum (seeFigure 7-47). The
rhodopsins absorb maximally at shorter wavelengths. Per- energy of a single photon of light is given by the Einstein
haps the freshwater environment makes sensitivity to the relation:
red end of the spectrum important.
Tracing the transduction of information from the ab-
sorption of a photon to the production of neuronal signals
leaves the question of how all this information about inci-
dent radiation is molded into a coherent view of the world where h is Planck's constant and v, c, and A are the fre-
unanswered. The collected information is passed on to quency, speed, and wavelength of light, respectively. Sub-
higher neuronal centers, where it is integrated and can stituting the values for a photon of blue light (A = 500 nm),
be used to shape behavior-a topic that is explored in the energy is calculated to be about 57 kcal-mol-l-
Chapter 11. almost 100-fold greater than the thermal energy. In vision,
detection is definitely not limited by thermal energy within
the detector. Instead, it has been found to be limited by the
LIMITATIONS O N SENSORY quanta1 nature of light itself.
RECEPTION In audition, the energy is given by the Einstein relation
To be most useful, a sensory receptor should be very sensi- for single phonons, which are quantum units of sound en-
tive to stimulation from the environment and should en- ergy analogous to photons of light. Animals hear sounds
code the information with perfect accuracy. In fact, no re- across a remarkably broad range of frequencies, from 10 to
ceptor meets these requirements, because of the physical lo5Hz. The energy of phonons at these frequencies ranges
properties of stimuli and of receptors; all receptors repre- from 7 x to 7 x erg. In the middle of this
sent compromises in how they receive and encode sensory range, the energy of a single phonon is 10 orders of mag-
information. Some physical principles that apply to recep- nitude (101°)below the limit of detection set by thermal en-
tors of many sensory modalities necessarily limit the fidelity ergy. This result indicates that the detection of acoustic
with which sensory information is received and transmit- stimuli is fundamentally limited by thermal noise, and there
ted by cells. In some cases, the accuracy of sensory recep- must be special mechanisms that permit auditory sensory
tion is limited by the relative magnitudes of the signals and reception. Indeed, some advantage is gained by tuning the
the background noise. This signal-to-noise ratio limits the detectors to limit their range, a common feature found in
performance of all systems that receive and transmit infor- the hair cells of auditory systems. Numerous mechanisms
mation, whether or not they are living. In other cases, the have evolved to combat the limitations of thermal noise,
performance and sensitivity of a sensory system are limited but direct measurements have also shown that sensory cells
by the form of energy to which the receptors are tuned. For in auditory systems faithfully reproduce the thermal noise
example, light is by its nature quantized into photons. No at their inputs.
receptor can receive less than one quantum of light, because As discussed earlier in this chapter, most chemical stim-
light does not exist in fractions of quanta. uli (olfaction, taste, chemotaxis) bind to specific receptors,
A major source of background noise arises from a rather than directly change ionic currents through mem-
corollary of the Third Law of Thermodynamics. That is, at brane channels. In this case, the relation between binding
all temperatures above O°K, molecules have kinetic energy energy and thermal energy determines the limits of detec-
and are in motion. Thermal energy is given by tion. The binding energies that have been measured in
-
chemical sensory systems are typically about 1kcal mol-I .
This energy is sufficiently greater than thermal energy that
chemoreceptors could theoretically count single molecules.
where k is Boltzmann's constant (1.3805 X 10-l6 erg K-l) There is, however, an important constraint dictated by the
and T is the absolute temperature. This equation gives the physics of receptor binding. The greater the binding energy,
energy that is associated with the movement of molecules the longer the molecule remains associated with its recep-
(i.e., Brownian motion) at an animal's body temperature. It tor. For a binding constant of lop6M, the association time
sets a lower limit on the sensitivity of receptors in detecting is about 3 X seconds; whereas, for a binding constant
signals because thermal energy provides a constant noise of lo-'' M (giving very high specificity), the association
level against which stimulation occurs. To detect an exter- would last for more than 5 minutes. Because the perfor-
nal signal, a receptor must be able to distinguish the signal mance of a receptor system depends at least in part on com-
from this baseline thermal noise. How easily can receptor parisons across many receptors, long binding times would
cells accomplish this task? require that comparisons be made over very long time pe-
Photoreceptors provide an example. At a body riods, and evolution seems to have shunned such a mecha-
temperature of 2S°C, the thermal energy is about nism. Instead, the binding constants between chemical
0.58 kcal-mol-', or 4 x 10-l4 erg. We must compare this stimuli and their receptor molecules are moderately high,
energy to the energy of a typical sensory stimulus. The stim- reducing the binding energy but also the time required to
ulus for a vertebrate photoreceptor is light in the visible transduce and interpret chemical signals.
270 P H Y S I O L O G I C A L PROCESSES
.........................................
Signal-to-noise properties can be predicted for stimuli dynamic range while retaining high sensitivity to weak
that activate electroreceptors and thermoreceptors. Elec- stimuli.
troreception is relatively widespread in aquatic organisms, Parallel inputs from receptors that cover different parts
in which it is used for navigation, communication, and pre- of the intensity range increase the range of stimulus inten-
dation. The energy in the electric fields, carried through wa- sities that can be perceived. The time-dependent loss of
ter at the frequencies used, is about 10 orders of magnitude sensitivity to a maintained stimulus, termed sensory adap-
below the thermal energy. Thus, the process of electro- tation, is a common property of receptor cells; some recep-
reception, like that of auditory reception, must be domi- tors adapt rapidly and others adapt slowly. The mecha-
nated by thermal noise in the detector. Thermoreception nisms responsible for sensory adaptation vary. Some take
depends on the detection of photons in the infrared region place in the receptor cell, others in the network of neurons
of the electromagneticspectrum (which has lower frequen- that carry the sensory information. In at least one case (the
cies and longer wavelengths than those in the visible spec- Limulus photoreceptor), adaptation results in part from in-
trum), and, by definition, it is limited by the temperature tracellular elevation of Ca2+, which blocks the light-
difference between the measured object and the measuring dependent activation of Na+- K+-selectiveion channels.
organ. Some animals, particularly beetles, have been found Some receptor cells exist individually, but others are or-
to perform at or near the theoretical limits; others have ap- ganized into sensory tissues and organs, such as the verte-
parent adaptations that keep their detectors cooler than the brate nasal epithelium or the retina of the eye. Anatomical
rest of the body, decreasing the background thermal noise. organization affects how a sensory organ functions. For ex-
As scientists have explored the limits of detection ample, the quality of the image formed by the vertebrate vi-
achieved by animal senses, it has become clear that many sual system depends on the presence of a lens and a huge
modalities operate at or near the theoretical limits imposed population of photoreceptor cells in the retina.
by physical laws. To accomplish this prodigious task, many Several sensory systems have features in common. In
types of receptors have evolved similar molecular mecha- particular, many receptor molecules contain seven trans-
nisms, and the mechanisms that are available to be used in memt>ranedomains, a feature that is also found in some
each sensory modality depend, at least to some extent, on neurotransmitter and hormone receptors. Many sensory
whether sensory reception is limited by thermal energy or systems also have common elements in the cascade of bio-
by the quanta1 nature of the stimulus. chemical events that immediately follow signal detection
and that amplify the signal.
Mechanoreception is a result of distortion or stretching
SUMMARY of the receptor membrane, directly producing changes in
Receptor cells are highly sensitive to specific kinds of stim- ion conductances. The deflection of the stereocilia of hair
ulus energy and relatively insensitive to other kinds of stim- cells provides directional information by modulating, up-
ulation, and they transduce the stimulus into an electrical ward or downward, the frequency of spontaneously oc-
signal, usually (but not always) a depolarization. The lower curring impulses of axons in the eighth cranial nerve. This
limit to sensation often depends on how much energy is function is the basis of reception in several sensory or-
carried in the signal compared with the energy in the ther- gans-the lateral-line system of fishes and amphibians,
mal noise within the organism. The transduction process is vertebrate audition, and the organs of equilibrium in both
most sensitive to weak stimuli, producing receptor signals vertebrates and invertebrates. The mammalian cochlea an-
that contain several orders of magnitude more energy than alyzes sound frequencies according to their effectiveness in
the stimulus itself. This sensitivity drops off with increasing displacing different parts of the basilar membrane, which
stimulus strength. In most receptor cells, the primary sites bears hair cells. Mechanical waves travel along the basilar
of reception and transduction are receptor molecules lo- membrane, set up by sound-driven movements of the ear-
cated in the cell membrane or in intracellular membranes. drum and auditory ossicles; they stimulate the hair cells,
Activation of receptor molecules causes the conduc- which in turn modulate their synaptic activity onto audi-
tance of particular classes of ion channels in the membrane tory nerve fibers. Certain sound frequencies stimulate each
to change; typically, the change in conductance permits the location along the basilar membrane more strongly than do
flow of a receptor current, producing a receptor potential. other frequencies, which is the basis for frequency discrim-
In many sensory modalities, the receptor cells do not them- ination in mammals.
selves produce APs. Instead, receptor potentials modulate Electroreceptors of fishes are modified hair cells that
the amount of neurotransmitter that the receptor cells have lost their cilia. Exogenous currents flowing through
release onto second-order neurons, which in turn initiates electroreceptor cells produce changes in the transmem-
or modulates the number of APs in second-order neurons. brane potential that modulate the release of transmitter at
Stimulus intensity is typically encoded in the frequency the base of the receptor cell, thus determining the rate of
of impulses, which in many sensory fibers is roughly pro- APs in sensory fibers.
portional to the logarithm of the intensity, up to a maxi- Visual receptors employ pigment molecules, in special-
mum frequency. The logarithmic relation between stimulus ized membranes, that undergo a conformational change af-
and response magnitudes permits reception over a large ter absorbing a photon. The change in the conformation of
the photoreceptive molecules initiates a cascade of reac- tensity encoded? How can a sensory system respond
tions that leads to achange in the conductance of the re- to stimuli whose intensity varies over many orders of
ceptor cell membrane. All visual pigments consist of a magnitude?
protein molecule (an opsin) combined with a carotenoid 7. Discuss three mechanisms that contribute to sensory
chromophore, either retinal (in rhodopsins) or 3-dehydro- adaptation.
retinal (in porphyropsins). The amino acid sequence of the 8. Discuss one example in which efferent activity can
opsin determines the absorption spectrum of each visual regulate the sensitivity of receptor cells.
pigment. A cis-trans isomerization of the carotenoid ini- 9. How are movements of the basilar membrane con-
tiates all visual responses. Absorption of photons is coupled verted into auditory nerve impulses?
to the opening (in invertebrates) or closing (in vertebrates) 10. Discuss the function of inner and outer hair cells in
of ion channels by intracellular second-messengers. In ver- the cochlea.
tebrate rods, photon capture by rhodopsin molecules leads How does spontaneous firing enhance the sensitivity
to the activation of associated G-protein molecules located of certain receptor systems-for example, lateral-
in the receptor membrane. Each G protein then activates line electroreceptors?
many phosphodiesterase molecules, each of which hy- How is the presence of an object perceived by elec-
drolyzes many molecules of the internal messenger cGMP. troreceptors of the weakly electric fishes?
In the dark, cGMP continually activates Na+ channels that What is the major difference between vertebrate and
carry the dark current. The light-dependent hydrolysis of invertebrate photoreceptor cells in their electrical re-
cGMP reduces the dark current, and a residual K+ current sponses to illumination?
hyperpolarizes the photoreceptor cell, reducing the steady Compare the mechanisms that allow the auditory
release of neurotransmitter at the inner segment. The re- system to distinguish the frequency of incident
duced rate of transmitter release causes a change in activity sounds and the visual system to distinguish the fre-
in the next higher-order neuron. quency of incident light.
Some vertebrates have three types of cones in the fovea, Outline the steps, as currently understood, in the
each containing a visual pigment that is maximally sensi- transduction of light energy in vertebrate visual
tive to a different part of the spectrum. The integration of receptors.
activity from all of these cones produces color vision. Rods, How does our current understanding of the physiol-
which in human beings all contain only one type of pho- ogy of color vision corroborate Young's trichromacy
topigment, are present in great densities in the periphery of theory?
the retina outside the fovea, are more sensitive than cones, Compare and contrast the morphological and func-
and show much greater synaptic convergence. As a result, tional properties of vertebrate rods and cones.
they exhlbit low acuity and high sensitivity. What allows some arthropods to respond to the ori-
entation of polarized light? Human beings cannot do
it. Why?
REVIEW QUESTIONS Compare the ways in which mammalian and teleost
1. Visual receptor cells can be stimulated by pressure, lenses focus images.
heat, and electricity, as well as by light, as long as the
intensity of these other stimuli is sufficiently great.
SUGGESTED READING
How can this fact be reconciled with the concept of
receptor specificity? Corey, D. P., and S. D. Roper. 1992. Sensory Transduction.
2. Choose one sensory modality and outline the steps 45th Annual Symposium of the Society of General
from energy absorption by a receptor cell to the ini- Physiologists. New York: Rockefeller University Press.
tiation of action potentials (APs) that will travel to A series of papers that discuss recent data from studies
the central nervous system. of transduction in many different sensory modalities.
3. Why must receptor potentials be converted into APs Dowling, J. 1987. The Retina. Cambridge, Mass.: Belknap
to be effective? Press of Harvard University Press. A very readable
4. All sensory information enters the central nervous compendium of information on the vertebrate retina
system in the form of APs having similar properties. written by a major contributor to our understanding of
How can we differentiate among various stimulus this sensory organ.
modalities? Finger, T. E., and W. L. Silver. 1987. Neurobiology of Taste
5. What is the difference between sensory transduction and Smell. New York: Wiley. A collection of papers
and sensory amplification? Choose one sensory considering the chemical senses in a broad range of
modality and describe how these two processes are animals.
related in that modality. Hudspeth, A. J. 1989. How the ear's works work. Nature
6. Discuss the relation between the intensity of a stimu- 341:397-404. A beautifully written account of audi-
lus and the magnitude of the signal sent to the central tory transduction by hair cells, written by a man who
nervous system by receptor cells. How is stimulus in- has played a major role in exploring the subject.
272 PHYSIOLOGICAL PROCESSES
.........................................
Kandel, E. R., J. H. Schwartz, and T. M. Jessell. 1991. Prin- Land, M., and R. Fernald. 1992. The evolution of eyes.
ciples of Neural Science. 3d ed. New York: Elsevier. Ann. Rev. Neurosci., 15:l-29. A consideration of the
An enormous and authoritative compendium of infor- physical and optical properties of visual organs across
mation about the function of the nervous system, from all of animal phylogeny.
the biophysics of membrane channels to the physiolog- Shepherd, G. M. 1994. Neurobiology. 3d ed. New York:
ical basis of memory and learning. Several chapters Oxford University Press. A concise text that considers
consider sensory mechanisms, with some emphasis on several sensory modalities in both vertebrates and
vertebrates. invertebrates.
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8
GLANDS: MECHANISMS
A N D COSTS OF SECRETION
cells secrete material into their surrounding envi- In this chapter, we first describe the nature and mecha-
ronment, either to form a protective barrier around nism of cellular secretions. We then move to the organ level,
the cell, or to communicate with other cells. Cells secreting discussing the adrenal medulla, the mammalian salivary
similar substances (e.g., hormones) are often collected to- gland, and the spinnerets glands of spiders that produce the
gether to form glands. The specialized cells composing a silk for webs. Because glandular secretions are involved in
gland act as a unit, secreting and sometimes excreting ma- all aspects of physiology, various glands are discussed
terial within the body or onto the body surface. Every ani- throughout this book; only a few examples are described in
mal has a large number of different glands, which vary in detail in this chapter. Finally, a discussion of the energy ex-
both structure and function. The particular types of glands pended in glandular activity rounds out this chapter.
possessed by an individual vary not only with the species,
but also during various developmental stages. The venom
glands of snakes, human sweat glands, wax glands of in- CELLULAR SECRETIONS
sects, the thyroid gland, and the pituitary are just a few of A surface coat is secreted by most cells. Mucus is also se-
the wide variety of glands found in the animal world. Glan- creted onto the external surface of epithelia, giving rise to
dular secretions are synthesized by cells that form the se- the term mucosal referring to the external surface of the ep-
cretory part of the gland and are released from the gland in ithelium. The surface coat allows cells to recognize each
response to an appropriate stimulus. The nature and extent other; the coat and mucus also form a protective barrier
of the secretion and the form of the stimulus vary greatly around epithelial cells, creating a somewhat controlled mi- -
among different glands. croenvironment between these cells and the surrounding
The secretions from glands constitute an important re- extracellular space. In addition, cells secrete various signal-
sponse of animals to a variety of situations. Feeding, for ex- ing substances that are used for communication between
ample, results in the massive activation of a whole range of cells. Such cellular communication varies from intimate
digestive glands (see Chapter 15).In vertebrates the secre- contact to distant signaling.
tion of hydrochloric acid (HCl) by cells lining the stomach
into the stomach lumen after eating can be so large as to Types and Functions of Secretions
cause a marked increase in blood pH, the so-called post- Secretions involved in communication between cells can be
prandial (after-dinner) alkaline tide. (Remember, an in- categorized by the distance at which they have an effect
crease in pH corresponds to more alkaline conditions.) This (Figure 8-1):
increase in blood pH can be very large, especially in car-
nivorous animals like crocodiles, which are capable of eat-
ing a whole gazelle at a single sitting! The webs spun by spi-
. Autocrine secretion refers to a secreted substance that
affects the secreting cell itself. An example is norepi-
ders, which are used to catch prey, are another example of
nephrine released from adrenergic nerve endings, which
glandular secretions. The nature of the secretion and the inhibits further release of norepinephrine from that
pattern of the web vary with the species and the environ-
nerve.
mental conditions. Mucus nets secreted by deep-sea fish
may play a role similar to that of spiders' webs in the inter- Paracrine secretion refers to a substance that has an ef-
actions between predator and prey in the unique environ- fect on neighboring cells. For example, during the in-
ment of the deep sea. Secretions from glands often play im- flammatory response localized vasodilation is induced
portant roles in mating behavior as well as reproduction in mainly by histamine released from mast cells in the area
general (see Chapter 9). of tissue damage.
274 PHYSIOLOGICAL PROCESSES
...............................................................................
Bloodstream -. 0
?
T~~~~~
cells 0000 *
Endocrine secretion refers to a substance that is released female silkworm moths. In certain marine invertebrates,
into the bloodstream and acts on a distant target tissue. such as clams and starfish, spawning of eggs and sperm is
Exocrine secretion refers to a substance that is released triggered by pheromones liberated along with the gametes.
onto the surface of the animal, including the surface of Thus, the spawning of one individual triggers spawning in
the gut and other internalized structures. others of both sexes. The adaptive value of such epidemic
r spawning is that it enhances the probability that sperm and
egg will meet and that fertilizationwill occur. A steroid that
Some exocrine secretions, called pheromones, are pro- induces molting in crabs also serves as the female sex at-
duced by one animal to communicate with another and are tractant, producing behavioral responses in males at con-
involved in initiating a range of physiological responses. In- centrations in the seawater as low as 10-l3 M. These sig-
dividual members of many animal species utilize pher- nals are an essential part of communicating about mating.
omones to communicate with each other. Among insects, Pheromones also are used to repel -predators. A corn-
-
for instance, pheromones function as a means of identify- mon example is the foul-smelling musk that makes skunks
ing the members of a colony. Pheromones also play an im- unpalatable to their enemies. A pair of scent glands near the
portant role in reproduction for many species. For exam- anus produce this yellowish, oily, malodorous secretion,
ple, bombykol is a powerful sex attractant released by which has been detected by humans at sea, as far as 30
kilometers from the nearest land. The repulsive odor is due The oligosaccharides within the glycocalyx can be seen us-
to trans-2-butene-1-thiol, 3-methyl-1-butanethiol and ing lectins (e.g., concanavalin A) that have been labeled
trans-2-butenyl methyl disulfide in the volatile portion of with fluorescent dyes or electron-dense material. Visualiza-
the scent. The encapsulating muscles close to the anal open- tion of the glycocalyx clearly shows that it protects the cell
ing are capable of propelling the musk over a meter. membrane and creates a microenvironment around the cell,
It is not uncommon for hormones, or their breakdown thereby modulating filtration and diffusion processes. Col-
products, to act as pheromones. Water in which ovulated lagen in the cell coat serves as a mechanical support for tis-
female goldfish have lived contains reproductive hormones sues and offers surfaces on which cells may slide.
and/or their metabolites, which can induce male goldfish The chemical composition of the glycocalyx also per-
sexual behavior; that is, these substances cause the male mits certain cells to recognize each other and adhere, form-
goldfish to become much more active and investigate tank- ing organized structures. For example, red blood cells have
mates. Likewise, steroid glucuronides (e.g., estradiol glu- specific cell-surface antigens that are distinguished by their
curonide) released by zebra fish shortly after ovulation at- terminal carbohydrates and form the basis of the ABO
tract males. Mixtures of steroid glucuronides, however, are blood groups. Red blood cells in the same blood group do
much more potent than single chemicals in eliciting male not aggregate, whereas those in different groups will ag-
sexual behavior in fish. gregate when mixed. In most tissues, similar cells aggregate
Some animals produce secretions that act both locally to form organs, and cells in culture will aggregate with
and at a distance and, therefore, have autocrine, paracrine, other like cells. Interestingly, reaggregation of chick em-
and endocrine effects. For instance, calcitonin produced in bryonic tissue is organ rather than species specific.
the gills of Pacific salmon modulates calcium flux across the The glycocalyx around some cells contains muco-
gills through calcitonin receptors in the gills; this calcitonin p~l~saccharides that can associate with proteins to form
produced by, and acting on, the gills acts as both an au- mucoproteins. Mucoproteins have a much longer polysac-
tocrine and paracrine secretion. Pacific salmon also pro- charide component than glycoproteins, are amorphous,
duce calcitonin in the ultimonbranchial gland; in this case, and form gels able to hold large amounts of water. The jelly
the calcitonin is released into the blood and subsequently coats of frog eggs are a common example of such gels. Al-
acts on the gills as an endocrine secretion. though distinct from the glycocalyx, mucus also contains
In addition to communication, secretions have numer- mucoproteins with a large number of sialic acid termini and
ous other functions. Saliva produced in the mouth helps creates a protective environment around the cell.
food slide down the esophagus, and pancreatic secretions Mucus is produced by specialized cells, called goblet
aid digestion. Snails secrete mucus with special elastic prop- cells, found in most epithelia. These cells secrete large
erties that enable them to slide and stick as they move over amounts of mucus, which can cover the surface of many
the ground. The path of a snail is marked by its secreted cells. For example, when a hagfish is disturbed, glands in its
mucus, which is essential to this mode of locomotion. skin produce enormous volumes of mucus; once released
Thus cells secrete material that can be detected, in some from the body, the mucus expands very rapidly in water
cases, by an adjacent cell or, in other instances, by other an- and can completely fill a bucket containing the hagfish
imals at a distance of 30 kilometers. Cells producing au- within minutes. This slimy coat presumably protects the
tocrine and paracrine secretions may be, but are not al- hagfish from attack. The mammalian lung provides a clear
ways, collected together to form glands. Cells secreting example of the protective role of mucus. The airways to the
hormones or pheromones almost always occur together in lungs continually secrete a sticky mucous layer that is pro-
glandular structures. As discussed below, many different pelled towards the mouth by the action of millions of tiny
cells produce a glycocalyx and mucus. Secretion of a wide cilia. The mucus traps dirt particles and bacteria; this mix-
variety of other substances (e.g., hormones, pheromones, ture of mucus, dirt, and bacteria is then carried away from
and digestive juices) varies from cell type to cell type. Many the lung surfaces by ciliary action and, subsequently, is ei-
of the substances secreted by different species have the same ther swallowed or expectorated. This production and
or similar functions, and these substances frequently have movement of mucus, along with antibodies secreted into
identical or related structures. In other words, there is a the mucus and alveolar macrophages, keeps the lungs clean
conservatism in the primary structure of secreted chemicals and free from infection. Tobacco smoke inhibits both cil-
(Spotlight 8-1). iary activity in the airways and the action of the alveolar
macrophages, but it enhances the production of mucus.
Surface Secretions: The Cell Coat and Mucus The "smoker's cough" is an attempt to remove the accu-
The external surface of the plasma membrane of all cells is mulated mucus.
protected by a cell coat, or glycocalyx, which is secreted by
the cell and is continuously renewed. The cell coat is com- Packaging and Transport of Secreted Material
posed of glycoproteins and polysaccharides, with nega- Cells that secrete specific substances are generally morpho-
tively charged sialic acid termini. The glycocalyx can be vi- logically polarized; that is, the synthesis and packaging of
sualized using appropriate dyes such as Alcian blue for light the secreted substance take place in one part of the secre-
microscopy and ruthenium red for electron microscopy. tory cell and its secretion takes place in another part
276 P H Y S I O L O G I C A L PROCESSES
...........................................................................
S P O T L I G H T 8-1 whereas GnRH induces the release of luteinizing hormone (LH),
which causes ovulation in humans (see Chapter 9). When in-
SUBSTANCES WITH jected into mice, yeast a-factor causes release of LH, but it has
a lower affinity for GnRH receptors than native GnRH (i.e., GnRH
SIMILAR STRUCTURES AND from mice). In most but not all cases, a native hormone has a
FUNCTIONS SECRETED BY higher affinity for it's own receptors and is more effective than a
similar but foreign substance.
DIFFERENT ORGANISMS Occasionally, a foreign substance has a greater effect than a
native hormone. This is true of mammalian calcitonin, which is
Just as organisms have many b~ochem~cal
pathways In common, produced by the C (clear) cells of the thyroid gland. Calcitonin
, there are many s ~ m ~ l a r ~ In
t ~ the
e s chemical structure of sub- acts on bones, the major body store of calcium, t o lower extra-
t stances secreted by d~fferentan~mals.Substances w ~ t h
s~m~lar cellular calcium levels under conditions when the calcium levels
funct~onsthat are secreted by quite d~fferentorganisms often are raised (see Chapter 9). It accomplishes this by inhibiting 0s-
have s ~ m ~ lstructures
ar For example, a-factor secreted by yeast teoclastic (bone-resorbing) activity without affecting osteoblas-
and gonadotrop~n-releasinghormone (GnRH) produced In the tic (bone-building) activity. Salmon and eels also produce a struc-
pltu~tarygland of mammals are small p e p t ~ d ehormones wlth turally similar calcitonin hormone in their ultimonbranchial
s~milaramino acid sequences N o t only are t h e ~ structures
r ho- glands. This foreign hormone is many times more effective than
mologous, but both secret~ons funct~on In reproduct~ve native human calcitonin in preventing bone resorption and thus
processes The a-factor acts as a matlng pheromone In yeast, lowering blood calcium in humans.
100
Native
li!L!L
25
100-11
10-5 10-3
(Figure 8-2).The nature of the synthesis and storage varies are similar in many respects to synaptic vesicles, which are
with the nature of the substance being secreted. For exam- somewhat smaller (-50 nm in diameter).
ple, the steroid hormones appear to be secreted in diffuse
molecular (i.e., unpackaged) form (see Chapter 9). Most
substances, however, are packaged in membrane-bound Polymer gels
vesicles within the secretory cell, later to be liberated into Mucus, a polymer gel, is packaged and stored along with
the extracellular space. Thus, electron microscopy of most various other chemicals as compact (condensed) granules
secretory tissues reveals membrane-limited secretory gran- in secretory vesicles within goblet cells. Mucus consists of
ules (secretory vesicles), 100 to 400 nm in diameter, that extremely long mucoprotein strands with a large number
contain the substance to be secreted. The terms secretory of sulfate and sialic acid termini, which are negatively
granule and secretory vesicle are used interchangeably, de- charged at neutral pH (Figure 8-3).Individual mucin chains
pending on whether the emphasis is on the contents (gran- are linked end-to-end by disulfide bonds between cysteine
ule) or the limiting membrane (vesicle).Secretory vesicles residues, creating extremely long mucin strands up to 4 to
Fldugh
endoplasmlc
Outer nuclear
membrane
]]I((( Dlsulf~debond
O g l y c o s ~ d ~bond
c
.N-acetylgalactosam~ne
~Threonlne
Monosacchar~des V S e r l n e
retlculum
Figure 8-3 Mucus IS a polymer gel conslstlng of mucoproteln strands
Figure 8-2 Secretoryprotelns aresyntheslzedin the rough endoplasmlc jolned end-to-end by dlsulflde bonds The dlsulflde bonds do not form
retlculum (ER), transferred In vesicles to the Golg~complex, and released cross-bridges,which would restrlct movement of the polymer chams No-
from the aplcal surface Afterthe protelns are concentrated In secretory tice the ollgosaccharlde slde chains, many of which have a negatively
vesicles, the vesicles move to and fuse wlth the apical surface membrane, charged slallc ac~dtermlnus In the condensed phase, the mucln strands
discharg~ngtheir contents Into the lumen of the gland by exocytosls form a hlghly tangled network [Adapted from Verdugo, 19901
6 pm in length. These mucin strands diffuse along their membrane so that the contents are expelled from the cell.
axes, rather like randomly moving snakes clustered in a In most cases, this process is regulated by the level of intra-
ball. Within secretory granules, the mucin strands form a cellular free calcium ions. Exocytosis appears to be the
highly condensed polymer network arranged as a tangled mechanism for releasing from cells all exocrine and en-
web. When released into water, however, the mucin net- docrine secretions that are stored in vesicles. In the exocy-
work can expand rapidly as much as several hundredfold tosis of mucus, a vesicle moves to the cell surface and a pore
and be diluted infinitely. forms as the vesicle fuses with the surface membrane. In-
Polymer gels can exist in two phases: a condensed or an crease in conductance of the pore has been shown to be in-
expanded hydrated phase. They can undergo phase tran- dependent of and, therefore, not caused by expansion of
sition from the condensed to the expanded phase and vice the polymer gel. Ion exchange occurs between the contents
versa very rapidly, similar to the change from a liquid to a of the vesicle and the extracellular space, causing the cal-
gas phase as water boils. High concentrations of calcium cium level to fall and perhaps the pH to rise within the vesi-
ions or hydrogen ions (i.e., low pH) can neutralize the neg- cle. As a result, the release of mucus by exocytosis is ex-
ative charges on the sialic acid termini of mucin strands, plosive, the mucus expanding so rapidly that it pops out of
thus favoring the condensed phase. Mucin vesicles appear the vesicle like the release of a jack-in-the-box (Figure
to contain sufficient calcium to keep the mucin in the con- 8-4A). The mucin network in the giant granule of the slug
densed phase. The low pH of mucin vesicles may also play can expand as much as 600-fold in 20 to 30 ms. This ex-
a role in the condensation process, and lipids within the tremely rapid rate of swelling is driven by repulsive forces
vesicles may help to ensure that the mucin network remains between the negative charges uncovered by the loss of cal-
in the condensed phase while stored in the vesicle. Calcium cium and perhaps the rise in pH, rather than by the diffu-
is not the only shielding cation that keeps polymers in the sion of water into the mucus, a process too slow to explain
condensed phase in vesicles. In chromaffin cell granules, for the rapid expansion that occurs (Figure 8-4B). It appears
example, chromogranin is the polymer and catecholamines that mucus is present in many different types of secretory
act as the cation, and in mast cell granules, heparin is the vesicles and probably assists in the release of stored mate-
polymer and histamine is the cation. rial under most conditions. Chromogranin probably plays
Mucus is released by exocytosis, which entails fusion of this role in chromaffin granules, aiding in the release of cat-
the membranes surrounding secretory vesicles with the cell echolamines into the blood.
278 P H Y S I O L O G I C A L PROCESSES
...............................................................................
Mucin strand
with membrane
Exocytosis of vesicle
0 5 10
Time (sec)
Figure 8-4 Mucus explodes from vesicles like a jack-in-the-box. lease of the vesicle contents t o the extracellular space. Water enters and
(A) Model for product release by exocytosis. Following fusion of thevesi- enlarges the vesicle as the mucus swells. (B) Time course of the swelling
cle to the plasma membrane, the shielding cations (n+)are releasedfrom of mucus in the extracellular space immediately after its release from a
the mucus inside the vesicle and/or extracellular anions (n-) flow inward. goblet cell in vitro. The change in radius as a function of time follows first
The net result is that the negative charges in the condensed polyionic order kinetics. [Adapted from Verdugo et al., 1987.1
mucin become unshielded, driving a fast volumetric expansion and re-
The length of the polymer strands in the tangled web of the nature of the fluid lining the airways. The abnormally
a condensed polymer gel affects its rate of expansion: the thick mucus found in humans suffering from cystic fibrosis
shorter the strands, the more rapid the rate of expansion. has its origins in defective ion transport processes across the
Some polymer networks (but not mucin networks) have co- epithelium of the lung airways, which changes the ionic
valent disulfide bonds linking the strands, which limit ex- composition of the extracellular fluid and, therefore, the
pansion and may influence exocytotic release. The most im- rheological properties of the mucus.
portant factor determining the rate of expansion of the
mucin gel, however, is the nature of the environment into Secretory and membrane proteins
which the mucin network is released. Hyperosmotic solu- The intracellular movement of secretory proteins has been
tions can inhibit swelling of the mucin granule, and the studied by pulse-chase radiography, a tracer technique in
ionic composition, pH, and quantity of fluid into which which radioactively labeled amino acids are incorporated
mucus is released has a marked effect on the final state of for a brief period into newly synthesized proteins. Such
hydration and ,therefore, the flow properties of released studies reveal that secretory proteins are synthesized on
mucus. For example, the properties of mucus secreted onto messenger RNA templates on the polyribosomes (poly-
the surfaces of the lung airways are strongly lnauenced by somes) of the rough endoplasmic reticulum (ER) and ac-
GLANDS: MECHANISMS A N D COSTS OF SECRETION 279
...............................................................................
1
Trans
region
sitional elements; the membrane of these elements buds off,
dncapsulating the secretory products in transfer vesicles
(Figure 8-5A).These transfer vesicles then migrate to the
Golgi complex, which consists of a stacked set of slightly
IMedial
region
concave, nearly flat membrane saccules with closely asso-
ciated free vesicles and vacuoles (Figure 8-SB).Microscopic
studies indicate that the membranes of the transfer vesicles
fuse with Golgi saccules. Within the Golgi complex, which
contains enzymes bound to the luminal membrane sur-
i
faces, some proteins undergo alterations such as the addi-
tion of sugar residues or the excision of fragments and join-
ing of two polypeptide chains.
Cis
region
The Golgi complex consists of at least three sets of cis-
ternae-the cis, medial, and trans cisternae. The cis face of
sacs
1 the Golgi complex receives transfer vesicles from the ER,
whereas the trans face or the trans-Golgi network (TGN)
B O Transfer vesicles f r o m t h e rough ER produces secretory vesicles, which subsequently move to
the surface of the cell (seeFigure 8-5B).It is believed that in
Figure 8-5 lntracellularvesicles transport secretory and membrane pro-
a process that begins in the Golgi cisternae, but which takes
teins. (A) Electron micrograph of the Golgi complex and rough ER in an
exocrine pancreatic cell. Notice the stacked layers of the Golgi complex
place mainly in the condensing vacuoles, water is drawn
and a forming secretory vesicle, as well as the transfer vesicles that shut- out of the future secretory vesicle, with the result that the
tle secretory and membrane proteins from the rough ER to the Golgi effective concentration of the enclosed protein increases 20-
complex. (B) Three-dimensional model of the Golgi complex and intra- to 25-fold. Mature secretory vesicles eventually reach the
cellular vesicles. Transfer vesicles that have budded off from the rough ER
plasma membrane to await the appropriate signal to release
fuse with the cis membranes of the Golgi complex. The secretory vesicles
that bud off from sacs on the trans membranes store secretory and mem-
their contents to the cell exterior.
brane proteins in concentrated form. [Part A courtesy of G. Palade; part Intracellular vesicles not only transport secretions to the
B from Lodish et al., 1995, after a model by J. Kephardt.] cell surface for exocytosis but also deliver proteins to be
280 PHYSIOLOGICAL PROCESSES
.........................................
incorporated into the plasma membrane. After synthesis in
the rough ER and transport to the Golgi complex, such
membrane proteins are incorporated into the vesicular
membrane and then inserted into the plasma membrane
when the vesicular membrane fuses with the plasma
membrane of the cell. This vesicular system is capable of di-
recting specific membrane proteins to different regions of
the secretory cell. For example, Na+-K+ATPase is delivered
to the basolateral membrane, and proton ATPase to the
apical membrane of the same cell. Thus, differences be-
tween the apical and basal regions of secretory cells are
maintained by the specific proteins delivered by vesi-
cular transport and incorporated into the surface mem-
branes.
The TGN is responsible for directed delivery of mater-
ial to the apical or basolateral surface. Some vesicles pro-
duced by the TGN migrate to the apical surface, whereas
other vesicles migrate to the basolateral surface. Newly syn-
thesized membrane proteins transported from the rough
ER to the Golgi complex are not yet sorted by ultimate des-
tination, but within the TGN they are sorted into vesicles
destined for either the apical or basolateral membrane
(Figure 8-6). Some proteins delivered to the basolateral
membrane are ultimately delivered to the apical membrane;
such transport is referred to as transcytotic delivery. The
Figure 8-6 The trans-Golgi networkflGN) sorts newly synthesized mem-
microtubular system appears to play a central role in the
brane proteins into vesicles destined for the apical or basolateral mem-
movement of vesicles to the cell surface, but the mechanism brane. After their synthesis in the rough ER, apical and basolateral
of sorting is not understood. membrane proteins move in common transfer vesicles to the Golgi com-
plex (I), where they occupy the same compartments. In the TGN, the api-
cal and basolateral proteins are sorted into ves~clesthat move, respec-
tively, to the apical membrane (2) and the basolateral membrane (3).
Some apically destined proteins first are delivered to the basolateral
membrane but then are retrieved and transported to the apical surface,
a process termed transcytotic delivery (4). Unlike secretory proteins re-
leased from the cell, membrane proteins are incorporated into the vesic-
ular membrane; they are inserted into the plasma membrane by fusion of
the vesicle with the plasma membrane.
A-
/ cell \
A Neuron 'l---'
B Neurosecretory cell
Y
GLANDULAR SECRETIONS The salivary glands are under direct neural control, influ-
enced by both conditioned and unconditioned reflexes. The
Glandular secretions result from the combined activity of a sight and smell of food can cause a marked increase in sali-
number of secretory cells. Secretion often occurs at a low vation, especially if the animal is hungry. In fact even the
"resting" level, which can be modulated up or down by sig- thought of food can increase saliva production. By associ-
nals acting on the gland. Some glands, however, exhibit no ating the ringing of bells with the appearance of food,
secretory activity until they are stimulated into action. For Pavlov was able to train dogs to salivate when bells were
instance, the nasal gland of birds may be inactive while a rung (see Spotlight 15-1).
bird drinks freshwater, but is activated to excrete salt fol-
lowing a drink of seawater. Various types of signals regu- Types and General Properties of Glands
late glandular activity: neurotransmitters released from Glands are classified as either endocrine or exocrine glands
neurons innervating the glandular tissue or hormones re- (Figure 8-9). Endocrine glands are organs that secrete hcr-
leased from other tissues. In addition, some glandular tis- mones directly into the circulatory system and modulate
sues respond directly or indirectly to conditions of the ex- body processes; for example, the thyroid gland produces
tracellular environment. For example, osmoregulatory thyroid hormone, which modulates growth. Endocrine
neurons in the vertebrate hypothalamus respond to the os- glands are sometimes referred to as the ductless glands. Ex-
motic pressure of the extracellular fluid bathing them, ocrine glands, on the other hand, secrete fluids through a
which, of course, reflects the osmotic pressure of the blood. duct onto the epithelial surface of the body; for example,
A Exocrine alands 1 2
Experimental
groups
Treatment
I-\
\-I
I I-\
\_I
Results
Secretion
Figure 8-9 Glands can b e divided into two broad structural types. (A)An thing that conditioned the blood and that the blood then
exocrlne gland, which releases secretionsvia a duct onto an epithelial sur- acted on the cockerel to produce male characteristics.
face. The primaryfluid is formed by ion transportwith water following 0s- The experiments carried out by Berthold paved the way
motically. Mucin plus a variety of other compounds may b e added t o the for many other similar experiments in which the effects of
primary fluid by exocytosis. The resulting primary secretion may b e mod-
removing and then replacing an organ were observed in or-
ified by reabsorption of material as the fluid passes down the duct. (B) An
endocrine gland, which is ductless and releases secretions directly into der to demonstrate an endocrine function for that organ.
the bloodstream. Water-soluble secretions are released by exocytosis of Since Berthold's experiments were published, a large num-
'
secretory vesicles, whereas lipid-soluble secretions may leave secretory ber of endocrine and exocrine glands, along with the chem-
cells by diffusion. ical structure of their secretions and mode of action, have
been described in detail. Although endocrine and exocrine
glands usually can be distinguished by the presence (ex-
ocrine) or absence (endocrine) of ducts, there is no such
thmg as a typical gland.
sweat glands produce sweat for evaporative cooling, and
the gallbladder stores bile salts produced in the liver and ex-
cretes them into the gut via the bile duct.
Glands have been studied for centuries. Many symptoms
of endocrine dysfunction were well known long before en-
docrine tissues had been identified and the function of their
secretionsdetermined. The study of endocrinology probably
began in 1849 when A. A. Berthold reported his classical ex-
periments in which he showed that castrated cockerels had
small combs and wattles, showed little interest in hens or Exocrine glands are more easily identified than en-
fighting, and had a weak crow (Figure 8-10).If he replaced docrine glands because of their duct leading to the body sur-
the testes into the abdominal cavity, then the comb and wat- face. On the other hand, the various endocrine tissues are
tle were large and the cock crowed and showed normal male structurally and chemically diverse, and some contain more
behavior. Berthold speculated that the testes secreted some- than one kind of secretory cell, each elaborating a different
284 PHYSIOLOGICAL PROCESSES
..............................................................................
hormone. Endocrine glands exhibit no common morpho- In many exocrine glands proteins, hormones, or other
logic plan or distinctive gross morphologic feature (other substances are added to the primary fluid by exocytosis of
than a rich vascularization). For this reason, establishing un- secretory vesicles. For example, mammary glands of mam-
equivocally that a particular tissue suspected of having an mals produce a primary fluid to which various substances,
endocrine function actually does and locating the site of se- including hormones, are added before the milk is consumed
cretion of a hormone have proved difficult in some cases. by the young. In the salivary glands, amylase and glyco-
The asymmetricaldistribution of ion pumps on the api- proteins are added to the primary fluid by exocytosis, as
cal and basolateral surfaces of secretory cells makes it pos- discussed in detail later. In some other exocrine glands, such
sible for cells to pump ions from one side to the other. The as the sweat glands, the primary fluid does not contain
movement of ions is followed by water. In many exocrine much in the way of additives. Unlike exocrine glands, most
glands (e.g., the rectal gland of sharks, the nasal gland of endocrine glands release hormones directly into the blood-
birds, and the sweat glands of mammals), ions are trans- stream without the formation of a primary fluid.
ported into the lumen with water following osmotically, In the following sections, one endocrine gland-the
yielding a fluid secretion, termed the primary fluid.Sodium adrenal medulla-and two exocrine glands-the salivary
chloride often is the secreted salt, but occasionally potas- gland and silk gland-are described in detail. Although
sium chloride is used. In some glands (e.g., sweat glands) presented as examples, each of these glands, like all glands,
some of the salt is reabsorbed in the duct leading from the has its own special properties; thus they should not be con-
secretory portion of the gland (see Figure 8-9A). sidered as typical or characteristic of glands in general.
TABLE 8-1
Vertebrate endocrine glands and tissues
Adrenal gland
Steroldogenlc tlssue (cortex) Aldosterone f Sodlum retention
Cortlsol and cortlcosterone f Carbohydrate metabol~smand sympathetic functlon
Chromaffln tlssue (medulla) Eplnephrlne and noreplnephrlne Multlple f and J, effects on nerves, muscles, cellularsecret~ons,
and
metabollsrn
Gastrointestinaltract Cholecystokinin f Secretion of enzymes by pancreatic acinar cell; f gall bladder
contraction
Chyrnodenin f Secretion of chymotrypsinogen from the exocrine pancreas
Gastric inhibitory peptide 1 Gastric acid (HCI) secretion
Gastrin f Gastric acid (HCI) secretion
Gastrin-releasing peptide f Gastrin secretion; J, gastric acid (HCI) secretion
Motilin f Gastric acid secretion and motility of intestinal villi
Neurotensin Enteric neurotransmitter
Secretin f Bicarbonate secretion by pancreatic acinar cells
Substance P Enteric neurotransm~tter
Vasoactive intestinal peptide f Intestinalsecretion of electrolytes
Heart (atrium) Atrial natriuretic factor f Salt and water excretion by kidney
Calcitriolt f Blood Ca2+,bone formation, and intestinal absorption of CaZ+
and
Erythropo~et~n (erythrocyte- f Production of red blood cells (erythropoies~s)
stlmulatlng factor)
Ovary:
Preluteal foll~cle Estradlol f Female sexual development and behav~or
Corpus luteum Progesterone f Growth of uterine lining and mammary glands, and maternal
behavior
Relaxln f Relaxat~onof publc symphys~sand dllatlon of uterlne cervlx
Pancreas (islets of Langerhans) Glucagon f Blood glucose, gluconeogenesis, and glycogenolysis
Insulin 1 Blood glucose; f protein, glycogen, and fat synthesis
Pancreatic polypeptide f J, Secretion of other pancreatic islet hormones
* f means hormone stlrnulates or Increases lndlcated effect, 1 means hormone lnhlblts or decreases tndlcated effect
tThe flnal steps In synthesls of calcltrlol from vltamln Dj occur In the k~dney,but the skln and llver also play a role In ~tssynthesls.
Endocrine Glands Identifying and studying endocrine tissue
Table 8-1 lists the major endocrine glands and tissues in As noted already, the absence of discrete mor~hO1Ogic
vertebrates, the hormones produced by each, and their markers complicates the identification of endocrine glands.
physiological role. (The pituitary hormones and details of The following criteria have been used to establish whether
hormonal regulation and action are discussed in Chap- a tissue has an endocrine function:
ter 9.) Endocrine tissues often are embedded in organs with
nonendocrine functions. For example, cells within the atria Ablation (removal) of the suspected tissue should pro-
of the heart produce atrial natriuretic peptide; this hormone duce deficiency symptoms in the subject. Experimen-
is released into the bloodstream in response to factors such tally this may be difficult to demonstrate if the tissue is
as a rise in venous pressure and helps regulate blood vol- part of an organ that has more than one function (e.g.,
ume. Although the role of the atria of the heart in blood cir- atrial tissue of the heart).
culation has been known for centuries, its role in the pro- Replacement (reimplantation)of the ablated tissue else-
duction of atrial natriuretic peptide was realized only where in the body should prevent or reverse the defi-
recently. Some hormone-like substances, including the ciency symptoms. If the effects produced by removal of
prostaglandins and leukotrienes, are produced by all or the tissue are due to the absence of a blood-borne sub-
nearly all tissues. Others, including some growth factors stance produced by that tissue, replacement of the ab-
and the endorphins, are produced by various selected lated tissue, which restores the missing hormone,
tissues. should restore normal function. Misleading results,
*Ang~otens~nogen and circulates In the bloodstream, where ~tIS cleaved by renln to form the actlve hormone anglotens~nII
IS produced In the l~ver
however, may be obtained when ablation-and-reim- amount of antibody. Unlabeled hormone will compete with
plantation experiments are done with tissues closely as- and therefore reduce the extent of binding of the labeled
sociated with the nervous system, because of the inter- hormone. Thus the quantity of hormone in a sample can be
ruptions of neural connections. determined by the extent to which it reduces the binding of
The deficiency symptoms should be relieved by replac- labeled hormone to the antibody. The development of RIA
ing the suspected hormone by injection. Successful re- has resulted in many new insights into the synthesis, secre-
placement is the most important criterion for identifi- tion and function of many hormones and many other sub-
cation of a suspected endocrine tissue and its hormone. stances. The use of monoclonal antibodies, which recognize
It is also the basis of replacement therapy for patients only one antigen, has improved the accuracy of detecting
with a dysfunctional endocrine gland. and quantifying hormones and their receptors by RIA.
Endocrinologists also use recombinant DNA tech-
Following purification of the suspected hormone, the niques in various ways. For instance, genetic material can
chemical structure of the active substance is deter- be inserted into bacteria to produce strains capable of syn-
mined. The molecule is then synthesized and tested for thesizing human hormones. Foreign genes have also been
biological potency. introduced into mammalian embryos; for example, when
Immunohistochemistry can be used to determine the the structural gene for rat growth hormone is introduced
cellular location of the hormone in different tissues into mouse embryos, the resulting mice grow much larger
once it has been isolated. than normal.
A variety of techniques have resulted in the rapid de- Mammalian adrenal medulla
velopment of endocrinology over the last two decades. For The paired mammalian adrenal glands, as the name im-
example, radioimmunoassays (RIAs) permit detection of plies, are situated near the kidneys, one attached to the su-
specific hormones in minute concentrations with a high de- perior end of each kidney (Figure 8-11).Each adrenal gland
gree of accuracy. Antibodies are raised against the hormone is in fact two glands in one: an outer layer, the adrenal cor-
in question, usually in a rabbit. A standard curve is then tex, surrounds an inner portion, the adrenal medulla
constructed to describe the binding of the hormone to the (Figure 8-12). The two portions of the mammalian adren-
antibody, using a radiolabeled hormone and a known als are of quite different origin. The cells of the cortex are
Abdominal aorta
Adrenal medulla
Medial adrenal
Figure 8-1 1 The adrenal glands in mammals are attached at the rostra1 medulla. Thus hormones produced in the cortex and released into the
ends of the kidneys. Two arteries enter the glands through the capsule blood are carried into the medulla, which is drained by the inferior
and branch into smaller vessels, which pass into the centrally located phrenicvein.
GLANDS: MECHANISMS A N D COSTS OF SECRETION 287
...............................................................................
Figure 8-12 Mammalian adrenal glands
have a recognizable cortex and medulla,
which produce different hormones. This
Adrenal
light micrograph reveals the outer cap-
medulla
sule, the three concentric layers of the cor-
tex, and the underlying medulla. Thezona
glomerulosa, the outermost cortical layer,
secretes mineralocorticoids;the zona fas-
Zona ciculata and the zona reticularis secrete
reticularis glucocorticoids. The adrenal medulla se-
cretes two catecholamines, epinephrine
and norepinephrine.[Courtesyof Frederic
H. Martini.]
Zona Adrenal
' fasciculata cortex
Zona
1 glomerulosa
: Capsule
- derived from mesodermal tissue, whereas those of the typical postganglionic sympathetic neurons. They are pre-
medulla are derived from epidermal tissue. The adrenal vented from doing so by the presence of high concentra-
cortex produces steroid hormones involved in blood ion tions of glucocorticoid hormones released from the sur-
and glucose regulation and anti-inflammatory reactions rounding cortex into the blood flowing from the cortex to
(see Chapter 9). the medulla (see Figure 8-11).
The cells of the adrenal medulla, on the other hand,
produce catecholamines, namely, epinephrine and norepi- Synthesis of catecholamines The production and release of
nephrine. Epinephrine and norepinephrine released from catecholamines including epinephrine and norepinephrine
sympathetic nerves and the adrenal medulla have numerous is outlined in Figure 8-13. The secretory granules within a
cardiovascular and metabolic effects, which in total consti- single chromaffin cell contain either norepinephrine or ep-
tute the fight-or-flight reaction. For example, plasma epi- inephrine, and each cell secretes one or the other cate-
nephrine levels can be elevated in a cat when it hears a dog cholamine. The granules also contain enkephalin, ATP, and
bark. This fight-or-flightreaction, or syndrome, is a response several acidic proteins called chromogranins. The cate-
to stress in which various tissues are activated and the body cholamines within the granule are probably bound to these
is mobilized to either attack or flee from the object of stress. chromogranins, which are polymers maintained in the con-
Catecholamines are not simply released during fight-or-flight densed state by the shielding action of catecholamines
' situations, but are released under a wide variety of physio- within the granule. Once a pore is opened in the vesicle, the
logical conditions, for example, during heavy exercise or even catecholamines begin to diffuse out and the chromogranin
when humans move from a sitting to a standing position. polymer rapidly expands, propelling the contents of the
Adrenal medulla cells are referred to as chromaffin vesicle into the extracellular space.
cells because they stain easily with chromium salts. The Norepinephrine is synthesized from tyrosine, with dopa
chromaffin cells that produce norepinephrine have dark- and dopamine as intermediate compounds (Figure 8-14).
staining irregular granules, whereas those that produce ep- Conversion of tyrosine to dopamine occurs in the cytosol,
inephrine have light-staining, spherical granules. Chro- catalyzed by tyrosine hydroxylase and dopa decarboxylase,
maffin cells are modified postganglionic sympathetic which are cytosolic enzymes. Dopamine is then incorpo-
neurons. A small number of cells within the medulla, which rated into the granules and converted to norepinephrine;
are somewhat intermediate between chromaffin cells and this reaction is catalyzed by dopamine P-hydroxylase
neurons, are referred to as small-granule chromaffin cells. (DBH)contained in the secretory granules. Norepinephrine
Under some conditions the chromaffin cells will grow into is methylated to epinephrine, a reaction catalyzed by
288 PHYSIOLOGICAL PROCESSES
...............................................................................
phenylethanolamine N-methyl transferase, which is found sociation of the adrenal cortex and medulla in mammals is
in the cytosol. Thus, norepinephrine must leave the of functional significance. In those tissues where there is
secretory granules to be converted to epinephrine, which close association of the chromaffin cells (adrenal medulla)
then reenters the granules (see Figure 8-13). with steroidogenic tissue (adrenal cortex), as in the mam-
Although chromaffin tissue and steroidogenic tissue are malian adrenals, most of the chromaffin cells produce epi-
associated in the adrenal glands in mammals, this is not the nephrine. As noted already, blood entering the medulla has
case in all vertebrates. In fish, for instance, the chromaffin passed through the cortex and thus carries high levels of
tissue is separate from the steroidogenic cells, but both are glucocorticoid hormones (see Figure 8-11).In the medulla
still in the general region of the kidney; the chromaffin tis- these glucocorticoids promote the synthesis of phenyl-
sue is associated with blood vessels, whereas the steroid- ethanolamine N-methyl transferase, the enzyme that cat-
producing cells are embedded in the kidney. The close as- alyzes conversion of norepinephrine to epinephrine.
O n the other hand, when chromaffin tissue isolated Effects and regulation of catecholamines Epinephrine and
from the influence of steroidogenic tissue, as in the dogfish, norepinephrine bind with adrenergic receptors, also termed
it produces more norepinephrine than epinephrine. The hu- adrenoreceptors, in cell membranes. This binding then ac-
man fetus has some isolated chromaffin tissue, which con- tivates one of a number of intracellular second messengers,
tains norepinephrine rather than epinephrine, presumably leading to a particular tissue response. (These pathways are
because of the absence of the action of steroidogenic tissue. described in detail in Chapter 9.) In a paper published in
Postganglionic sympathetic nerves also produce norepi- 1948, R. P. Ahlquist concluded that there are two types of
nephrine for the same reason, that is, the absence of a adrenoreceptors-a and p-that differ in their sensitivity
marked influence of steroid hormones. to sympathetic amines. More recent studies have demon-
strated several subtypes of both a- and P-adrenoreceptors
Release of catecholamines The release of epinephrine and based on the ability of various drugs to either activate or
norepinephrine from the adrenal medulla is controlled block receptor activity (Figure 8-16).
by the action of preganglionic sympathetic nerves (Figure The a,-adrenoreceptors mediate smooth muscle
8-15). These preganglionic fibers are cholinergic; that is, contraction in many tissues. Stimulation of these recep-
they release acetylcholine as a neurotransmitter. When the tors results in activation of the inositol trisphosphate
chromaffin cells are stimulated by acetylcholine, their mem- (InsP;) pathway, leading to elevation of intracellular InsP,
brane conductance for Ca2+increases, resulting in an influx (Figure 8-17).Elevated InsP, causes release of calcium from
of Ca2+and elevation of intracellular Ca2+levels; this rise stores within the cell; the resulting rise in cytosolic calcium
in intracellular Ca2+ in turn causes the further release of causes muscle contraction (see Chapter 9). There is evi-
both epinephrine and norepinephrine by exocytosis (see dence that there are subtypes of a,-adrenoreceptors in dif-
Figure 8-13). Catecholamines cause an increase in blood ferent tissues. The a,-adrenoreceptors located in presynap-
flow to the adrenals, and this effect also augments cate- tic cells at noradrenergic synapses cause inhibition of
cholamine release from the adrenal medulla. Thus the re- norepinephrine release, an action mediated by an inhibitory
lease of catecholamines has a positive feedback on further effect on adenylate cyclase. Thus, these receptors are part
catecholamine release. (The release of norepinephrine from of a short negative-feedback loop in which the release of
postganglionic sympathetic nerves, however, inhibits fur- norepinephrine inhibits further release of norepinephrine.
ther norepinephrine release from these nerve endings. In This is sometimes referred to as autoinhibition. There are
this case, negative feedback operates.) ATP is stored in the also a,-adrenoreceptors located on some postsynaptic sites
granules of chromaffin cells and released along with cate- in liver, brain, and some smooth muscle.
cholamines. ATP and its breakdown product adenosine, The p-adrenoreceptors also are divided into two sub-
which inhibit release of catecholamines by reducing cal- types, PI- and p2-adrenoreceptors, both of which activate
cium influx, provide negative-feedback control on cate- adenylate cyclase, leading to an increase in cAMP (see Fig-
cholamine release from the medulla. Hypoxia also stimu- ure 8-17). Stimulation of PI-adrenoreceptors, largely due
lates catecholamine release from chromaffin cells. When to neuronal release of norepinephrine, results in increased
chromaffin cells are not innervated (e.g., those located in contraction of cardiac muscle and the release of fatty acids
the hagfish heart), hypoxia is an important stimulus for cat- from adipose tissue, whereas stimulation of p2-adrenore-
echolamine release. ceptors, largely due to elevated levels of circulating cate-
Catecholamines released into the extracellular fluid are cholamines, mediates bronchodilation and vasodilation.
rapidly taken up and either stored in secretory vesicles or The elevation of cAMP resulting from stimulation of P,-
destroyed by monoamine oxidase located on the outer adrenoreceptors increases calcium conductance, thereby
membrane of mitochondria (see Figure 8-13). Catechol- raising the intracellular calcium level, which in turn aug-
amines in the extracellular space are catabolized by cate- ments muscle contraction. In contrast, elevation of cAMP
cholamine-0-methyltransferase, especially in the liver and following stimulation of p,-adrenoreceptors causes activa-
kidney, and the breakdown products are excreted. The ac- tion of the calcium pump rather than an increase in calcium
tual level of catecholamines circulating in the blood thus conductance. Thus, following p2-adrenoreceptor stimula-
depends on the balance between their release, uptake, and tion, calciunl is both sequestered within and extruded from
catabolism. Although the level of catecholamines in the the cell, so intracellular calcium levels fall, promoting mus-
blood is dominated by release from the adrenal medulla, re- cle relaxation.
lease from postganglionic sympathetic nerves contributes The following generalizations can be made concerning
significant amounts to blood levels. Adrenergic nerves re- the roles of adrenoreceptors:
lease norepinephrine, whereas the medulla releases mainly
epinephrine, so the relative activity of nerves and the a-Adrenoreceptors mediate smooth muscle contraction
medulla will also influence the relative levels of epinephrine (except in intestinal smooth muscle) and (with a few ex-
and norepinephrine in the blood. Catecholamine levels in ceptions) inhibit cellular secretion.
the blood can remain elevated for only a few minutes in hu- @-Adrenoreceptors mediate smooth muscle relaxa-
mans, but they can remain high for several hours following tion (except in cardiac muscle) and stimulate cellular
exhaustive exercise in fish. secretion.
290 PHYSIOLOGICAL PROCESSES
......................................
O z - y - N H 2
COOH
H 0 G ! - ( 7 H - N H 2
COOH
Tyroslne
hydroxylase
HO-(~-E-~H-NH~
COOH
L-aromatlc ammo
acid decarboxylasa
Dopamine
P-hydroxylase
Phenylethanolamlne
N-methyltransferase(glucocorticoidsT)
Figure 8-14 The catecholamines-dopamine, norepinephrine,and ep- ethanolam~neN-methyltransferase and, therefore, promote conversion
inephrine-are synthesized from phenylalanine and tyrosine. Glucocor- of norepinephrine to epinephrine.
ticoids produced by the adrenal cortex increase the activity of phenyl-
Higher
center
;ympathetic Catecholamine
ganglion
producing cell
spinhl
cord
Epinephrine or
norepinephrine
Figure 8-15 Hormone secretion by the adrenal medulla is regulated by then synapse on the catecholamine-producing cells. Acetylcholine lib-
neural stimuli. Sympathetic nerve axons originating in the spinal cord erated from these preganglionic nerve endings stimulate the secretion
pass through the sympathetic ganglia without synapse formation, but of medullary hormones.
G L A N D S : M E C H A N I S M S A N D COSTS O F SECRETION 291
...............................................................................
Practolol Butoxamine
(@,-antagonist) @-antagonist)
lsoproterenol
(p-agonist)
Prenalterol Salbutamol
(B,-agonist) (p,-agonist)
Figure 8-16 A varlety of drugs can actlvate (agon~sts)or block (antago- ceptor subtypes and t o determ~nethe effects of catecholam~neson d ~ f -
n~sts)adrenoreceptors. These drugs have been used t o ~dent~fy
adrenore- ferent tlssues.
Epinephrine or norepinephrine
/ \
f
InsP3+ DAG
I
Figure 8-17 Binding of catecholamines to a,-, a,-,P,-, or p,-adrenore- second messenger and the latter involves inositol trisphosphate (InsP,)
ceptors either activates (+) or inhibits (-) a second-messenger pathway. and diacylglycerol (DAG) as second messengers. See Figures 9-1 1
Adrenoreceptor signal transduction occurs via the aden~latecyclase or and 9-13 for more details. [Adapted from Hadley, 1992.1
the membrane phospholipid pathway. The former involves CAMPas a
292 PHYSIOLOGICAL PROCESSES
.......................................
If P-adrenoreceptors cause relaxation in a tissue, then cholamines. In the estrogen-dominated uterus, stimulation
cholinergic eceptors promote contraction in that same of p-adrenoreceptors causes contraction, whereas it causes
tissue. relaxation during pregnancy.
If a-adrenoreceptors cause contraction in a tissue, then In summary, then, the action of catecholamines depends
cholinergic receptors normally regulate relaxation in on the rate and site of their release (i.e., from the adrenal
that same tissue. medulla or nerves) and the rate of uptake andlor break-
down of the catecholamines once released. In addition, the
The physiological action of catecholamines is quite vari- type and distribution of receptors on target tissues, and the
able and is influenced by other factors. For example, neu- up- or down-regulation of these receptors due to past expe-
ropeptide Y (NPY), which sometimes is co-released with rience, have a marked effect on the nature and magnitude of
norepinephrine from adrenergic nerves, modulates the ac- the response. The presence or absence of steroids can influ-
tion of catecholamines on the InsP3second-messengerpath- ence both adrenoreceptor density and the levels of enzymes
way, augmenting the action of catecholamines in some tis- involved in the conversion of norepinephrine to epinephrine;
sues and reducing their action in other tissues. Many other the latter effect modulates the ratio of these two compounds
factors can modulate both the release and action of cate- released into the blood. The level and nature of gonadal
cholamines. Adenosine, for instance, has been shown to in- steroids, for example, can alter the response of the uterus to
hibit catecholamine release from the bovine adrenal medulla catecholaminesfrom contraction to relaxation. Finally other
by reducing calcium flux. Adenosine is released from tissues substances, such as ATP, adenosine, and neuropeptide Y, can
during hypoxia, but it is rapidly destroyed in the blood, so modulate the release and action of catecholamines. As a re-
its effects are limited to the region of production. sult of these various modulating mechanisms, physiological
Alterations in adrenoreceptor density in the membrane responses to epinephrine and norepinephrine vary widely
of target cells also can modify catecholamine action. An in- depending on both the tissue in question and the physiolog-
crease in receptor density is referred to as up-regulation; a ical state of the animal (Table 8-2).
decrease, as down-regulation. Continual exposure to cate-
cholamines can lead to down-regulation of receptor con-
centration and, therefore, a decreased response. Sympa-
thetic denervation can lead to up-regulation of receptors
and, therefore, to increased sensitivity of a tissue to circu-
lating catecholamines. Adrenoreceptor density also can be
affected by other substances, in particular steroid hor-
mones. Glucocorticoidsnot only modulate adrenoreceptor
density but also have other effects on the action of cate- Exocrine Glands
Unlike endocrine secretions, the output of an exocrine
gland does not ooze into the circulation but generally flows
TABLE 8-2
Physiological responses t o epinephrine and norepinephrine
through a duct into a body cavity (e.g., the mouth, gut,
nasal passage, or urinary tract) that is in continuity with the
Response to
exterior. As noted earlier, exocrine secretions usually are
Variable Epinephrine Norepinephrine aqueous mixtures, consisting of a water-based primary
Heart rate Increase Decrease*
fluid and added components, rather than a single sub-
Cardiac output Increase Variable
stance. In the alimentary canal, these mixtures typically
consist of water, ions, enzymes, and mucus. Exocrine tis-
Total peripheral Decrease Increase
resistance sues of the alimentary canal include the salivary glands, se-
Blood pressure Rise Greater rise
cretory cells in the stomach and intestinal epithelium, and
Respiration Stimulation stimulation
secretory cells of the liver and pancreas.
An exocrine gland typically consists of an invaginated
Skin vessels Constriction Constriction
epithelium of closely packed secretory cells lining a blind
Muscles vessels Dilatation Constriction
cavity called the acinus (see Figure 8-9A). Several acini con-
Bronchus Dilatation Less dilatation
nect to a small duct that, in turn, connects to a larger duct
Metabolism Increase Slight increase leading to the lumen of the digestive canal or to some part
Oxygen Increase Slight effect of the body surface. The basal surfaces of the epithelial cells
consumption
are usually in close contact with the circulation. Once the
Blood sugar Increase Slight increase
primary secretory products are free in the acinar lumen,
Kidney Vasoconstriction Vasoconstriction they generally become secondarily modified in the secretory
*This effect is secondary to peripheral vasoconstrictionthat raises blood duct. This modification can involve further transport of wa-
pressure. In the isolated heart, norepinephrineincreases the rate.
ter and electrolytes into or out of the duct to produce the fi-
Source: Bell et al., 1972. nal secretory juice.
Exocrine glands (e.g., sweat glands) are classified as TABLE 8-3
apocrine or eccrine glands based on their structure. An ec- 1n0'9anic constituents of whole saliva ("'9.100 m1-l)
crine gland has a coiled, unbranched duct that leads from Constituent Range Mean
the secretory region; the duct opening lies perpendicular to Sodium 0-80 15 resting
the body surface. Eccrine glands respond to elevated tem- 60 stimulated
peratures by secreting a clear fluid that evaporates and Potassium 80
cools the body. An apocrine gland has a branched duct Calcium 6
leading from the secretory region to the surface. Apocrine Phosphorus (inorganic) 17 resting
glands often produce a turbid or white secretion,which can 12 stimulated
be released by an apocrine, merocrine, or holocrine secre- Chloride -
tory mechanism, not simply by apocrine secretion, as the Thiocyanate 9 (smokers)
name of these glands might suggest. This extensive, and of- 2 (nonsmokers)
ten confusing, terminology is a reflection of the vast array Fluoride (parts/106) 0.03 resting
of glands with a multiplicity of functions found in animals. 0.01 stimulated
Bicarbonate 6 resting
36 stimulated
Vertebrate salivary gland
-
The saliva present in the human mouth is a complex mix- - - -
Figure 8-18 Production and release of the primary secretion by salivary lumen. Exocytosis of amylase and glycoproteins stored in secretory gran-
gland acinar cells is under neural control. Stimulation of a-adrenorecep- ules is promoted by activation of the adenylate cyclase (AC) pathway due
tors (a-R), acetylcholine receptors (Ach-R), and substance P receptors to stimulation of receptors for vasoactive intestinal peptide (VIP-R) and
(SP-R)activates phospholipase C (PLC).This enzyme splits phosphatidyl 0-adrenoreceptors (P-R). DAG and increased cytosolic Ca2+also pro-
inositol biphosphate (PIP2)into diacylglycerol (DAG) and inositol trisphos- mote exocytosis.The primary secretion released into the acinus lumen is
phate (InsP,), leading to release of stored Ca2+and opening of potassium modified as it passes through the salivary gland dud. See text for further
channels. As a result of various ion movements, NaCl and water enter the discussion. [Adapted from Edgar, 1992.1
and C1- enter the cell. These movements of Na+ and K+ are rises, with increased flow. The addition of bicarbonate to
counteracted by a Na+-K+ATPase, which maintains Na+ the fluid must somehow be coupled to the flow rate, such
and K+ levels in the cell. Thus, Na+ and K+ are cycled that an increase in flow promotes bicarbonate addition to
through the membrane, and the only net transfer is the in- the duct fluid.
ward movement of C1- ions, which move across the cell '- --
and leave via the apical (luminal)membrane. That is, there Invertebrate silk gland
is a net movement of C1- across the acinar cell from the The number and variety of glands in invertebrates is prob-
blood to the lumen of the gland. This generates a transep- ably larger than that in vertebrates. The silk gland is de-
ithelial potential that is positive on the blood side and cre- scribed here not so much because it is representative of a
ates the driving force for diffusion of Na+ through para- large number of invertebrate exocrine glands, but because
cellular channels from the blood to the lumen. This it is reasonably well understood. Many insects and spiders
movement of NaCl into the lumen establishes the osmotic produce silk threads from silk glands to make webs and
gradient generating the flow of water into the lumen. spin cocoons. The silkworm, Bombyx mori, is raised com-
Binding of norepinephrine to P-adrenoreceptors or of mercially for its larvae, which spin a protective cocoon.
vasoactive intestinal peptide to peptidergic receptors acti- Each cocoon produced by a pupating larva consists of
vates the adenyl cyclase pathway (see Figure 8-18).This re- about 275 m of silk thread. Commercial silk thread is made
sults in the formation of CAMP,which in turn activates a by weaving threads from several cocoons together.
protein kinase that stimulates exocytosis. Diacylglycerol, The production of silk cloth began in China some 4000
formed in the phospholipase C pathway, also promotes ex- years ago. Silk brought to Europe over the Silk Road from
ocytosis of amylase, mucous glycoproteins, and proline- China was used to make robes for Roman emperors. Silk
rich glycoproteins into the lumen. cloth was worn by the horsemen of Genghis Khan for pro-
The primary secretion, which consists of water, sodium tection, because arrows do not penetrate silk easily and ar-
chloride, amino acids, proteins, and glycoproteins, is forced rows can be removed by pulling on the silk thrust into the
into the salivary duct by the formation of more fluid. As it wound. This strong light material was one of several rea-
passes down the duct, potassium bicarbonate is added to sons for the great military success of the Mongols. Silk from
the fluid and some sodium is reabsorbed. Because less the silk moth is still used to produce fabric for today's con-
sodium is reabsorbed at high flow rates, the final product sumers. Although spider silk is stronger than that of the silk
leaving the duct approaches the composition of the primary moth, there has been little or no commercial use of the silk
secretion during heavy salivation. In contrast, the bicar- made by spiders; certainly European knights never went
bonate level in the final secretion does not fall, but in fact into battle covered by spider silk for protection!
Spider silk and webs Spiders are a very prolific group. It
has been estimated that an acre of meadow in England may
contain over two million spiders. One reason for the suc-
cess of this group is that the over 30,000 species of spiders
spin silk. Spider silk is made by spinneret glands on the un-
derside of the abdomen, which exude a liquid that hardens
into silk threads once it leaves the gland. These silk threads
are used as a dragline and to make a variety of webs, silken
egg cases, and silk-lined tunnels. The main, but not the
only, function of webs is to capture prey, such as insects and
other small animals, which become entangled in, or stuck
to, the web. The webs vibrate easily, and the spider can de-
tect the position and the nature of the animal in the web by
the pattern of vibration. Different behavioral responses are
evoked by different patterns of vibration of the web. A
male, wishing to be recognized as a potential mate rather
than food, vibrates the web in a species-specificpattern to
evoke the appropriate response from the female.
Not all spiders make webs. The large, nonpoisonous
tarantula spider, Lycosa tarantula, relies instead on its
speed to capture prey. Most spiders bite their captured prey,
using their fangs to inject poisons to subdue and digest their
victims, after which the spider sucks out the digested juices.
Only a few species of spider are dangerous to humans. The
male black widow spider, Lactrodectus mactans, is harm-
less, but the much larger female has a venomous bite; al-
though a bite causes pain and fever, human victims usually
survive. Female black widow spiders are about 1.3 cm long
and have a large red patch below the abdomen.
Spiders continually produce silk thread, which can trail
behind them as a dragline and is fastened at intervals to the
substratum. Spiders can become aerial by swinging from
their dragline attached to a bush or tree; bungie-jumping is
a common event in the daily life of many spiders. Spider
webs undoubtedly evolved from this dragline, with the sim-
plest webs being sticky threads hanging in the breeze to
catch insects. More complex webs are constructed in either
two or three dimensions and have intricate designs for snar-
ing prey. Webs are set up in the flight paths of insects and
other small animals. Some webs are vertical, whereas oth- Figure 8-19 Different species of spider build webs with a characteristic
ers are horizontal to catch insects flying up from the design. The common names of spiders often are based on the appear-
ground. In many webs, some of the threads break as the in- ance of their webs. [Adapted from "Spider Webs and Silks," by Fritz
sect hurtles into the web; the more the insect struggles, the Vollrath. Copyright O 1992 by Scientific American, Inc. All rights re-
served.]
more tangled it becomes in the web. In others, portions of
the web are sticky and the prey becomes attached to the
web through these sticky strands. Webs are designed so that a single spiral thread starting at the center and gradually
an insect flying into a web does not bounce out due to elas- spiraling outwards.
tic recoil, as in a trampoline.
Obviously the tensile properties of the threads and the
pattern of the web determines the characteristics of the he largest object caug
web. There are many web designs, and spider species are are the limitations to
often named for the type of web they construct: ladder-web
spiders, funnel spiders, filmy dome spiders, and mesh-web
spiders, to name just a few (Figure 8-19).A familiar web in
temperate climates is that of the garden cross spider, Ara- Silk production by spiders The abdominal silk glands are
neus diadematus, which spins a two-dimensional web with large and open via modified appendages, the spinnerets,
spokes radating from the center that are joked together by each of which has several spigots. The spinnerets are like
Y
4 Nanometers
conical gun towers and are very mobile. The threads, pro- valving mechanism of the spigot or by changing to another
duced in abdominal glands and extruded through spigots, gland, which enables spiders to produce silk for a wide va-
consist of alpha-keratin crystals embedded in a rubber-like riety of purposes (Figure 8-21). Spiders also coat the
matrix of amino acid chains, which are not cross-linked to threads with fungicides and bactericides, which prevent mi-
the crystalline alpha-keratin structures (Figure 8-20). In croorganisms from consuming webs. The presence of these
many cases the extruded thread is dry and may remain so substances probably accounts for the use of spider webs in
because of an oily lipid covering. These dry threads are elas- folk medicine to heal cuts and abrasions to the skin.
tic and have great strength, but they can only be extended Spiders expend a great deal of energy constructing
to about 25% of their length before breaking. The silk webs, which often are damaged rather quickly. Spiders will
threads are stiff and brittle when dry, but become pliable eat their own damaged webs, an important source of amino
when wet. acids, and construct new webs daily, often overnight. The
Dry threads in three-dimensional webs break when an garden cross spider, for example, can construct a web in less
insect collides with the web, thereby snaring the prey by than an hour using about 20 m of thread.
tangling it in a network of threads. In two-dimensional
webs, like that of the garden cross spider, dry threads form
the spokes of the web, but wet threads form the sticky con-
ENERGY COST OF GLANDULAR
tinuous spiral that forms the shape of the web. The spiral
ACTIVITY
thread has glue droplets surrounding glycoprotein dough- Glands can have very high rates of secretory activity. For
nuts at intervals along its length; these make the thread ad- example, the extra energy expended by a nursing mother
hesive so that insects stick to the web. The cribellate spi- for milk production can be equivalent to the energy expen-
ders, on the other hand, produce an adhesive dry thread by diture of a long-distance runner. In litter-bearing animals
covering it with a loose network of entangling amino acid the energy cost of lactation is even greater. Breast milk is the
chains, rather like Velcro. sole source of nourishment for newborn mice until they are
Each spider has several different silk glands, each of almost half the size of the mother mouse. For a litter of
which produces a unique silk characterized by the compo- eight, the total weight of the babies at weaning is four times
sition of its amino acid matrix. Spiders probably can adjust that of the mother. Thus the mother must eat enough food
the valving mechanism at the exit of the gland to create to supply all the nutrients for four times her body weight,
thicker threads. But if threads with a different amino acid 75% of which is diverted to lactation to supply the litter.
matrix are required, they use a different gland. Thus, the Food intake by lactating mice increases with litter size as il-
quality of the silk can be changed either by altering the lustrated in Figure 8-22.
Figure 8-21 Different silks having different functions can be produced switching from one gland to another, a spider can produce the silk ap-
by the same spider. The garden cross spider, A. diadematus, has seven propriate to the task at hand. [Adapted from "Spider Webs and Silks," by
different abdominal glands each of which produces silkwith a character- FritzVollrath. Copyright 0 1992 by Scientific American, Inc. All rights re-
istic amino acid matrix composition. The various glands open into com- sewed.]
mon spigots, but silk from only one gland is extruded at one time. By
Litter size The graphs in Figure 8-23 show that on average food
12 pups intake doubles in lactating ground squirrels during the
300 r Body mass of which is associated with sweating; the remaining one
third is associated with respiration. Evaporative heat loss is
increased greatly by sweating. The prodigious volume of
sweat produced during vigorous exercise or when it is hot
can be sufficient to prevent excessive heating of the body.
Humans can survive in a sauna at temperatures high enough
to cook meat because of the activity of sweat glands, which
produce enough sweat and, therefore, sufficient evaporative
200 1
0
I I
10
I I
20
I I
30
I I
40
cooling to maintain body temperature below 40°C. If the
humidity of the sauna is increased by pouring water on the
Age of pups (days)
heated rocks, evaporative cooling is decreased, leading to an
immediate rise in body surface temperature.
SUMMARY
Glands are organs composed of specialized cells that act as
a unit. Secretions are synthesized by cells that form the se-
cretory part of the gland. In response to an appropriate
stimulus, glandular secretions are released into the blood-
stream or onto the surface of an internal cavity or the body.
The nature and extent of secretions and the form of the
(basal rnetabol~sm) stimulus varies greatly among glands. There are a multitude
1 I I
0 10 20 30 40 of glands that vary not only among species but also during
Age of pups (days) development.
Most glandular secretions contain mucus, which is
Figure 8-23 Most of the increased energy intake by lactating ground
packaged in vesicles along with other secretory products
squirrels is stored as potential energy in the milk produced and thus is
transferred to the pups. Data are mean values for females producing an
and subsequently released from the gland by exocytosis.
average litter of four pups. (A) Maternal body mass as function of age of The contents of these vesicles often are released into a pri-
pups. Note that mothers did not gain weight despite their increasedfood mary fluid formed by the active transport of ions (e.g.,
consumption during the nursing period. (8) Comparison of maternal NaCI) followed by water into the lumen of the gland. Se-
food consumption, milk production, and basal metabolism, expressed as
cretory vesicles are formed in the Golgi complex and are di-
daily energy equivalents in kilojoules (k].d-'), during the nursing period.
Food consumption and basal metabolism of nonlactating control squir-
rected to either the apical or basal membranes of the secre-
rels are indicated by shaded bars. Note that milk production accountsfor tory cell by the trans-Golgi network. Exocytosis of vesicles
about 75% of the energy intake by mothers. [Adapted from Kenagy, usually is triggered by an increase in intracellular calcium
Stevenson, and Masman, 1989.1 levels resulting from neural or hormonal stimulation of the
secreting cell. In some cases secreting cells are stimulated by
environmental changes.
Glandular secretions often are critical for survival of an- Many secretions function in communication between
imals. For example, the sweat secreted onto the body sur- cells. Such secretions are classified into four types based on
face of mammals and some other mammals is important in the distance at which they exert an effect. An autocrine se-
temperature regulation. Evaporation of sweat from the cretion affects the secreting cell itself. A paracrine secretion
body surface dissipates heat and is one mechanism of heat has an effect on neighboring cells. An endocrine secretion
loss in mammals. The American statesman, politician, is released into the bloodstream and acts on a distant target
diplomat, newspaper publisher, and scientist, Benjamin tissue. Exocrine secretions used for communication, called
Franklin (1706-1790), who was elected a Fellow of the pheromones, are released through a duct onto the epithe-
Royal Society for his work on electricity, was also interested lial surface of the body; these secretions permit one animal
in temperature regulation of the body. He realized that skin to communicate with another. Some secretions can act both
temperature was lower than deep body temperature due to locally and at a distance and, therefore, have autocrine,
evaporative cooling of sweat. About 20% of the heat loss paracrine, and endocrine effects. Cells that produce au-
GLANDS: MECHANISMSAND COSTS OF SECRETION 299
...............................................................................
tocrine and paracrine secretions may be, but are not al- REVIEW QUESTIONS
ways, collected together to form glands. Cells that produce
endocrine secretions or pheromones are almost always col- 1. What role does mucus play in exocytosis? Explain
lected together into glandular structures. the rapid expulsion of mucus from vesicles associated
Glands can be characterized as either endocrine or ex- with exocytosis.
ocrine glands. Because endocrine glands lack any charac- 2. What is the role of the trans-Golgi network in deter-
teristic morphologic markers, a variety of techniques have mining cell polarity?
been used to identify such glands. The development of ra- 3. Discuss the differences between autocrine, paracrine,
dioimmunoassays (RIAs) and recombinant DNA tech- neurocrine, and endocrine, secretion. What are
niques has led to rapid developments in endocrinology over pheromones?
the past several decades. Exocrine glands are easier to rec- 4. What criteria must be met before a tissue can be
ognize than endocrine glands because they all possess a unequivocally identified as having an endocrine
duct and secrete material onto the body surface. function?
The adrenal medulla, an endocrine gland, secretes the 5. What is the significance of having the adrenal
catecholamines epinephrine and norepinephrine into the medulla and cortex collected together in a single or-
bloodstream. In the mammalian adrenal medulla the cate- gan? How does the circulatory pattern in the adrenal
cholamine-producing chromaffin cells are associated with glands affect the relative secretion of epinephrine and
steroidogenic tissue and most cells produce epinephrine. norepinephrine?
However, in some species (e.g., dogfish),chromaffin cells are 6. Explain how catecholamines can have so many dif-
not associated with steroidogenic tissue as in sharks and as ferent actions.
a result norepinephrine production dominates. Cate- 7. Explain how differential activation of sympathetic
cholamines have a large number of effects, both on the cir- and parasympathetic nerves can influence the com-
culation and metabolism. They act through both a- and position of saliva.
p-adrenoreceptors, which are linked to the inositol tris- 8. How does a spider alter the composition of silk pro-
phosphate and adenylate cyclase second-messengersystems, duced for making a web and other structures?
respectively. ATP, neuropeptide Y and adenosine can modu- 9. Describe the differences between dry and wet silk
late the release and activity of catecholamines. In addition, threads.
the density of adrenoreceptors on target tissues can be up- or 10. How much of the total energy budget of an animal is
down-regulated, thereby modulating catecholamine action. spent on secretion? How could this be estimated?
The products of exocrine glands flow through ducts 11. Discuss from an energetics point of view why many
onto the body surface. The surface may be enclosed as in the mammals give birth to young in the spring.
case of the mouth or gut. Salivary glands are exocrine glands
that secrete saliva into the mouth. Saliva is about 99.5% wa-
ter and contains a number of ions. Except for some hydrol-
SUGGESTED READINGS
ysis of polysaccharides into disaccharides by salivary amy-
lase, digestion in the mouth is minimal. Saliva serves as a Edgar, W. M. 1992. Saliva: Its secretions, compositions and
lubricant, assisting in eating, swallowing, and talking. It also functions. Br. Entomol. J. 172:305-312. (A concise de-
has an antibacterial action, which helps reduce tooth decay. scription of the functional organization of the mam-
Spiders have abdominal exocrine glands that produce malian salivary gland.)
silk threads, which often are used to form webs. These webs Hadley, M. E. 1992. Endocrinology. 3d ed. Chapters 1,2,
are made from both dry and wet silk threads and can have and 14. Englewood Cliffs, N.J.: Prentice Hall. (A use-
very complex designs. The tensile properties of the threads ful general text on endocrinology.)
and the pattern of the web determines the characteristics of Matsumoto, A., and S. Ischii, eds. 1992. Atlas of Endocrine
the web. Spider species are often named for the type of web Organs. Heidelberg: Springer-Verlag. (Beautiful dia-
they produce. Silk threads are extruded through spigots and grams of the structure of vertebrate endocrine organs.)
consist of alpha-keratin crystals embedded in a rubber-like Pimplakar, S. W., and K. Simons. 1993. Role of heterotri-
matrix of amino acid chains, which are not cross-linked to meric G proteins in polarized membrane transport.
the crystalline alpha-keratin structure. Different silk glands 1. Cell Sci. 17(Suppl):27- 32. (Plenty of information,
in a spider produce silk with a different amino acid compo- but not for the beginner.)
sition. A spider can change the characteristics of the silk pro- Verdugo, P. 1994. Molecular biophysics of mucin secretion.
duced either by altering the valving mechanism of the spigot In T. Takishima and S. Shimura, eds., Airway Secretion:
or by changing to another gland. Physiological Basis for the Control of Mucous Hyper-
Glands are critical for survival, but they can be expen- secretion. New York: Marcel Dekker, pp. 101- 121. (A
sive to maintain and operate. A lactating mouse must eat good review of the subject.)
enough food to supply nutrients for herself and her off- Vollrath, F. 1992. Spider webs and silks. Sci. Am.
spring, which may have a collective mass of four times that 266(3):70-76. (All you wanted to know about spider
of her own body before weaning. webs.)
,p,t:,::,,,; ,;: ~,:,~~~,~h1,1~~.iiI;~~~~:;I";~~tjr;;.1~~,~~~~~~,,~~,~,;:~,. . . * . . . ...I ir!;,3~!?:7~:~g4:.,.:,j.3~:1~::l;
:~~!;,{::t~... .. 'ii;i+:s~.li . ' Y .,.
: ~ :..,,..,, . ;?.. 'i..:,,'2.:.:?e.t,
:::
~:,j:~"r;~t,v:,
4
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b ,{it:
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,* , . .
1, , ,$:,
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9
HORMONES: REGULATION
1
I
AND ACTION
L
ment, the milieu interieur, that bathes the cells of the body.
0 ne of the great advances in evolutionary history was
the appearance of metazoa-multicellular organisms
in which different tissues specialized in different functions.
He concluded that animals became more independent of
the surrounding environment as they became more able to
This division of labor required that each type of tissue be control the composition of the internal environment
able to communicate with other types so as to coordinate bathing the cells. Walter Cannon (1871 - 1945), who
their activities to promote the survival of the organism. taught at Harvard University, coined the term homeosta-
(Table 9-1 summarizes the various types of messenger and sis to describe the tendency of the normal body to maintain
regulatory molecules found in metazoa.) steady states, especially the constancy of the milieu interieur
The French physiologist Claude Bernard (1813 - 1878) (see Chapter 1).Homeostasis is achieved by coordination
emphasized the differences between the external environ- of a complex set of physiological processes via chemical
ment that surrounds an animal from the internal environ- and/or electrical communication between tissues that
TABLE 9-1
Classification of chemical messengers and regulators
CAMP
lnositol triphosphate (InsP,)
Diacylglycerol(DAG)
Neurotransm~tters Nerve cells Synapt~ctransm~ss~on: transported short distances; Acetylchol~ne
brief durat~onof act~vity Seroton~n
Norep~nephr~ne
Neuromodulators Nerve cells Alteration of responses of ion channels to stimuli Norepinephrine
Neuropeptides
Neurohormones Nerve cells Endocr~nefunct~ontransported by c~rculat~on; Vertebrate neurohypophyseal
troplc effect common hormones
Arthropod developmental
hormones
Glandular hormones Nonneural endocrine Endocrinefunction: transported throughout body Epinephrine
tissues to distant target organs Ecdysone
Juven~lehormone
lnsulln
Local hormones Various tissues Endocrinefunction: paracrine actions on nearby Prostaglandins
targets H~stam~ne
Pheromones Glands openlng to lntraspec~f~c
commun~cat~on
between ~nd~v~duals Bombykol
the env~ronment
302 P H Y S I O L O G I C A L PROCESSES
........................................
elicits appropriate responses. Hormones play a central role Hormones are synthesized by specific tissues or glands.
in this communication and thus are critical to homeostasis. Hormones are secreted into the bloodstream, which
As discussed in Chapter 8, chemical signaling can in- carries them to their site(s)of action.
volve autocrine, paracrine, endocrine, or exocrine secre-
tions (see Figure 8-1). In each type of signaling, the target Hormones change the activities of target tissues or
cells bind the signaling molecules via special proteins called organs.
receptors, which are specific for a particular molecule. This
binding initiates the response of the target cell. We saw in Although hormone molecules come into contact with all
Chapter 6 that neurotransmitters are released from nerve the tissues in the body, only cells that contain receptors spe-
cells and act over short distances to activate receptors on cific for a particular hormone are affected by it. Binding of
postsynaptic cells, an example of paracrine action. In con- many hormones to their receptors triggers a cascade of two
trast, hormones released from various glands travel or more intracellular signaling molecules, called second
through the bloodstream to act on distance target cells, the messengers, that lead to a specific response in the target
prototype of simple endocrine action. tissue.
It should be noted that chemical regulation of cellular The amount of hormone produced by an endocrine
processes can be found in even the most primitive plants gland generally is small, and it is diluted in the blood and
and animal species, and undoubtedly preceded the origin of interstitial fluid. Thus hormones must be effective at very
the metazoa. For example, individual amoebas of the cel- low concentrations (typically between and 10-l2 M).
lular slime molds exhibit aggregating behavior in response By way of comparison, if human taste buds could detect
to CAMP, which is liberated by individual amoebas. (In sugar at 10-l2 M, we would be able to taste a pinch of
higher organisms, CAMPis a ubiquitous regulatory mole- sugar dissolved in a large swimming pool full of coffee or
cule, generally involved in intracellular signaling.) An even tea. [In contrast, the local concentrations of synaptic neu-
more primitive kind of chemical regulation occurs in the rotransmitters are much higher (-5 x M), and these
freshwater coelenterate Hydra. The water from a crowded substances are effective only at such concentrations.] The
culture of Hydra induces the differentiation of reproductive high sensitivity of hormonal signaling is due to the high
tissues in that animal. This effect is mediated by the ele- affinity of target cell receptors for hormones. As discussed
vated concentration of CO,, which accumulates as a nor- later, binding of a hormone molecule to its receptor leads to
mal by-product of metabolism. Thus, chemical regulatory a cascade of enzymatic steps that amplify the effect; thus
agents include relatively nonspecific molecules (e.g., NO, just a few hormone molecules can influence thousands or
CO,, H+, O,, and Ca2+)and the more complex messenger millions of molecular reactions within a cell.
molecules, produced specifically for mediating cellular
communication and regulation. Chemical Types and General Functions
This chapter focuses on the actions of glandular hor- of Hormones
mones and neurohormones. Hormones coordinate the Although hormones exhibit diverse chemical structures,
functions of animal tissues and organs on a time scale of most of the known hormones found in metozoans belong
minutes to days. The functions under hormonal control in- to one of four structural categories (Figure 9-1):
clude growth, maintenance, osmoregulation, reproduction,
and behavior, among others. Amines, including the catecholamines epinephrine and
norepinephrine, as well as the thyroid hormones are
small molecules derived from amino acids.
Prostaglandins (or eicosanoids) are cyclic unsaturated
hydroxy fatty acids synthesized in membranes from 20-
carbon fatty acid chains.
Steroid hormones (e.g., testosterone and estrogen) are
cyclic hydrocarbon derivatives synthesized in all in-
stances from the precursor steroid cholesterol.
Peptide and protein hormones (e.g., insulin) are the
ENDOCRINE SYSTEMS: OVERVIEW largest and most complex hormones.
William Bayliss and Ernest H. Starling described the first In contrast to neurotransmitters, which signal rapidly
hormone to be discovered, secretin, a substance liberated over short distances, hormones signal more slowly over
from the mucosa of the small ~ntestinethat causes Increased longer distances. Thus endocrine systems are well suited for
flow from the pancreas (see Chapter 15).Starling (1908)in- regulatory functions that are sustained for minutes, hours,
troduced the term hormone, derlved from the Greek for "I or days. These include the maintenance of blood osmolar-
arouse." He proposed that three characteristic properties ity (antidiuretic hormone) and of blood sugar (insulm),reg-
define hormones: ulation of metabolic rates (growth hormone and thyrox-
A Arnine B Prostaglandin
Prostaglandin PGE,
C Steroid D Peptide
S
1 Np NH2 NH2
I
NH2
? " Chain A
Gly Ile Val GluGlu Cys CysAlaSer Val CysSer LeuTyrGlu Leu Glu AspTyrCys Asp
1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21
s
I
PheValAspGlu HIS Leu CysGlySer HIS Leu Val Glu AlaLeuTyr Leu Val Glu Arg GlyPhePheTyrThr Pre Lys Ala
0 Chain 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 2 6 2 7 28 29 30
Testosterone Insulin (bovine)
Figure 9-1 Most hormones belong to one of four structural categories. hormones are lipid insoluble, whereas steroid hormones and pros-
Amine hormones (with the exception of thyroid hormones) and peptide taglandins are lipid soluble.
ine), control of sexual activity and the reproductive cycles hormone. Such feedback can involve either a short loop or
(sex hormones), and modification of behavior (various hor- long loop. In short-loop feedback, the concentration of the
mones). In fact, the rapid-acting activity of the nervous sys- hormone itself, or an effect produced by it, acts directly
tem and the slower, more-sustained activity of the endocrine back on the endocrine tissue to reduce secretion, thereby
system complement one another in the overall integration keeping hormone secretion in check. Long-loop feedback
of physiological and metabolic functions in a body. A given operates on similar principles, but it includes more elements
molecule may serve as a neurotransmitter in some in- in series.
stances, and the same or a closely related molecule may act When an extremely rapid response is required, the en-
as a hormone in other instances. In fact, there is an exceed- docrine tissue may be subject to positive feedback; that is,
ingly close and overlapping relationship between the ner- the secretion of a hormone leads directly or indirectly to its
vous and the endocrine systems. Indeed, in many respects increased secretion. Positive feedback is most common in
the nervous system can be viewed as perhaps the most im- the early phases of the response. For instance, positive feed-
portant endocrine organ, for it produces certain hormones back occurs early in the reproductive cycle of some verte-
that regulate the activity of many endocrine tissues. brates (and perhaps also invertebrates) when responses
(e.g., increase in level of luteinizing hormone) must peak
Regulation of Hormone Secretion relatively quickly. Ultimately, of course, this must be coun-
The secretion of hormones occurs generally at a resting level tered by a process that ends the rapid increase.
that is modulated up or down by signals acting on the en-
docrine tissue. These signals often are neurohormones,
which are released from specialized neurons and act directly
on the endocrine tissue, as discussed in the next section. In
some cases, the endocrine tissue responds directly to condi-
tions of the extracellular environment (e.g., changes in os-
molarity). Endocrine tissues are part of either feedforward
or feedback circuits. In a feedforward circuit, secretion is
not modulated by any consequences of the secreted hor-
mone, whereas in a feedback circuit, secretion is modulated
by one or more consequences of the secreted hormone.
The secretory activities of endocrine tissues generally
NEUROENDOCRINE SYSTEMS
are modulated by negative feedback (Figure 9-2 on page
304). That is, the increasing concentration of the hormone As discussed earlier, secretion of hormones from some en-
itself, or a response to the hormone by the target tissue (e.g., docrine tissues is regulated by neurohormones, which are
reduced blood glucose levels in the insulin loop), has an in- produced by specialized nerve cells called neurosecretory
hlbitory effect on either the synthesis or release of the same cells. Some neurohormones secreted from neurosecretory
Endocrine
Short loop
A A +
T~ssue
S~gnalfrom
tissue B
Signal from
tissue C
Figure 9-2 Most endocrlne tlssues are subject to negat~ve-feedback nal from secondarytarget t~ssues(C) control secretory actcvlty In an open
control In short-loopfeedback, the response of the primary target tlssue loop there IS no feedback
(B) feeds back onto the endocrrne gland. In long-loop feedback, a s~g-
cells in the hypothalamus regulate secretion of various and are carried in the bloodstream to target endocrine tis-
glandular hormones from the nonneural anterior pituitary sues or other target tissues.
gland. In contrast, the neurohormones released from the
posterior pituitary gland act directly on various target Of the Anterior
tissues; these hormones are produced in neurosecretory =land
cell bodies located in the anterior hypothalamus. This closk The axons of some hypothalamic neurosecretory cells have
relation between neural and endocrine systems is the basis their endings in the median eminence at the floor of the
of the neuroendocrine reflex (Figure 9-3). The neurosecre- hypothalamus (Figure 9-5). These cells secrete at least
tory cells in the hypothalamus respond to sensory input seven hormones that control secretion of various hor-
from various parts of the body. The pituitary gland, also mones by the anterior pituitary gland (Table 9-2). All but
termed the hypophysis, is a small appendage lying below one of these hypothalamic releasing hormones (RHs)and
the hypothalamus. Because it secretes at least nine hor- release-inhibiting hormones (RIHs)are peptides (Spotlight
mones, the pituitary gland has been called the "master 9-1). The discovery of these hypothalamic hormones has
gland. " proved to be one of the most important developments in
Although ordinary nerve cells and most neurosecretory vertebrate endocrinology, opening investigations into the
cells are generally similar, they exhibit several differences. orchestration of virtually the entire vertebrate endocrine
First, the secretory vesicles containing neurosecretory hor- system.
mones typically are 100-400 nm in diameter, whereas the As early as the 1930s, studies revealed that capillaries
presynaptic vesicles containing neurotransmitters in ordi- within the median eminence converge to form a series of
nary nerve cells are much smaller, 30-60 nm in diameter portal vessels that carry blood directly from the neurose-
(Figure 9-4).Second, although ordinary nerve cells use both cretory tissue of the median eminence to the glandular se-
slow and fast axonal transport systems, neurosecretory cretory tissue of the anterior pituitary gland. There they
cells appear to use only fast axonal transport, moving neu- break up again into a capillary bed before finally recon-
rohormones at rates of up to 2800 mm a day. Third, ordi- verging to join the venous system. This portal system en-
nary nerve cells form synapses with other cells at their hances chemical communication from the hypothalamus to
terminals, whereas neurosecretory axons generally termi- the anterior pituitary gland by carrying the hypothalamic
nate in clusters in a bed of capillaries, forming a discrete RHs and RIHs directly to the interstitium of the anterior
neurohemal organ (see Figure 9-3). The neurohormones re- pituitary gland, also known as the adenohypophysis (see
leased into the interstitial space diffuse into the capillaries Figure 9-5). Here these hypothalamic hormones come into
HORMONES: REGULATION A N D A C T I O N 305
...............................................................................
Stimuli acting
on nervous system
Hormone synthesis
contact with the glandular endocrine cells that secrete Glandular Hormones Released from the Anterior
adenohypophyseal hormones, either stimulating or in- Pituitary Gland
hibiting their secretory activity. Because of the direct portal The anterior lobe of the pituitary gland consists of the pars
connection from the hypothalamus to the anterior pituitary distalis, pars tuberalis, and pars intermedia. Six hormones
gland, very low concentrations of the RHs and RIHs can are released from the pars distalis and one from the pars in-
produce effects on the anterior pituitary gland. Once these termedia in mammals (see Figure 9-5). Although all the
hormones enter the general circulation, they are diluted to glandular secretory cells of the adenohypophysis are gen-
ineffective concentrations and are enzymatically degraded erally similar in appearance, they can be classified into two
within several minutes. histochemically distinct types:
ialar
\L Paraventricular
k Portal vessels
Hypothalamo-hypophyseal
neurosecretory tract
Posterior lobe
(neurohyphysis,
pars nervosa)
Anterior lobe
(adenohypophys~s)
pars distalis
7
,/
I
T-
vein
Growth hormone
Antidiuretic hormone
Prolactin pars (from supraoptic nucleus)
Thyrotropin intermedia oxytocin (from para-
Follicle-stimulating hormone
ventricular nucleus)
Luteinizing hormone
Adrenocorticotrooin
I Melanocyte-stimulating hormone
Figure 9-5 Hormonal secretion from the primate pituitary gland (hy- ing or release-inhibitinghormones secreted by hypothalamic neurose-
pophysis) is controlled by the hypothalamus.The anterior lobe of the pi- cretory endings in the median eminence are carried via the portal vessels
tuitary gland (adenohypophysis)consists of the pars distalis, pars inter- (hypothalamo-hypophyseal portal system)to the anterior pituitary gland,
media, and pars tuberalis. (The pars tuberalis, not shown, consists of a where they stimulate (or inhibit)secretion of several glandular hormones.
thin layer of cells surroundingthe pituitarystalk.)The posterior lobe (neu- Two neurohormones produced in hypothalamic cell bodies are released
rohypophysis), an extension of the brain, consists of neural tissue, from terminals of the neurosecretory cells in the posterior pituitary
whereas the anterior lobe consists of nonneural glandular tissue. Releas- glands.
Acidophils, which stain orange or red with acid dyes, cyte-stimulating hormone (MSH),luteinizinghormone
secrete growth hormone (GH; also termed somato- (LH),and follicle-stimulating hormone (FSH).
tropin) and prolactin (PRL).
Like ACTH, TSH, LH, and FSH are primarily tropic
Basophils, which stain blue with basic dyes, secrete in their actions (Table 9-3).That is, they act on other en-
ACTH, thyroid-stimulating hormone (TSH),melano- docrine tissues (e.g., thyroid, gonads, and adrenal cortex),
HORMONES: REGULATION A N D ACTION 307
TABLE 9-2
Hypothalamic hormones that stimulate or inhibit release of adenohypophyseal hormones*
Hormone Structure Primary action in mammals Regulation
Stirnulatory
Corticotropin-releasing Peptide Stimulates ACTH release Stressful neural input increases secretion;
hormone (CRH) ACTH inhibits secretion
GH-releasing hormone (GRH) Peptide Stimulates GH release Hypoglycemia stimulates secretion
Gonadotropin-releasing Peptide Stimulates release of FSH In the male, low blood testosterone levels
hormone (GnRH) and LH stimulate secretion; in the female, neural
input and decreased estrogen levels
stimulate secretion; high blood FSH or
LH inhibits secretion
TSH-releasing hormone Peptide Stimulates TSH release and Low body temperatures induce secretion;
(TRH) proladin secretion thyroid hormone inhlbits secretion
Inhibitory
MSH-inhibiting Peptide Inhibits MSH release Melatonin stimulates secretion
hormone (MIH)
Prolactin-inhibiting Amine Inhibits prolactin release High levels of prolactin increase secretion;
hormone (PIH) estrogen, testosterone, and neural stimuli
(suckling) inhibit secretion
Somatostatin Peptide Inhibits release of GH and many Exercise induces secretion; hormone is rapidly
(GH-inhibiting hormone, other hormones (e.g., TSH, inactivated in body tlssues
GIH) insulin, glucagon)
*ACTH = adrenocorticotropinhormone; FSH = follicle-stimulatinghormone; GH = growth hormone; LH = luteinlzlng hormone; MSH = melanocyte-
stimulating hormone; TSH = thyroid-stimulatinghormone.
regulating the secretory activity of these target glands. LH crease the synthesis and dispersal of melanin, leading to
and FSH, which act on the gonads, often are referred to as darkening of the skin.
gonadotropins. Thus the effect of these tropic hormones The relations between the hypothalamus and the ante-
on nonendocrine somatic tissues is indirect, operating rior pituitary gland are summarized in Figure 9-6. The three
through the hormones released from their target glands. release-inhibiting hormones from the hypothalamus sup-
The remaining adenohypophyseal hormones-growth press the release from the anterior pituitary gland of MSH,
hormone, prolactin, and MSH-are direct-acting hor- prolactin, and growth hormone. Growth hormone is also
mones; that is, they act directly on somatic target tissues under the control of a releasing hormone. Note the short
without the intervention of other hormones. The actions and long feedback loops, involving ACTH, TSH, FSH, and
of growth hormone and prolactin are discussed in later LH, which control the hypothalamo-anterior pituitary sys-
sections. MSH, whose release is regulated by MIH from tem and the long feedback loop, involving growth hormone,
the hypothalamus, acts on pigment cells in the skin to in- prolactin, and MSH, which controls the hypothalamus.
TABLE 9-3
Tropic hormones of the anterior pituitary gland
Adrenocorticotropin Peptide Adrenal cortex Increasessynthesis and CRH stimulates release; acth
(ACTH) secretion of steroid hormones slows release of CRH
by adrenal cortex
Follicle-stimulating Glycoprotein Ovarina follicles (female) In female, stimulates GnRH stimulates release; inhibln
hormone (FSH) Seminiferous tublues maturation of ovarian follicles and steroid sex hormones inhibit
male In male, increases sperm release
production
Lutelnlzlng hormone Glycoproteln Ovarlan lnterstltlal cells In female, ~nducesf~nal GnRH stimulates release, inhlbln
(LH) (female) maturat~onof ovarlan follicles, and sterold sex hormones lnhlblt
Testlcular lnterstlt~alcells estrogen secretion, ovulation release
(male) corpus luteum format~on,and
progesterone secretlon
In male, Increases synthesls
and secretlon of androgens
Thyrold-stimulating Glycoproteln Thyroid gland Increases synthes~sand TRH lnduces secretion, thyrold
hormone (TSH) secret~onof thyrold hormones hormones and somatostatin
slow release
Growth hormone
MSH (pars intermedia)
ACTH
TSH
FSH
Somatic
hormones
Prolactin
LH II
Somatic
nonendocrine
target
metabolites
Metabolic
Neurohormones Released from the Posterior The amino acid sequences of mammalian oxytocin
Pituitary Gland and arginine vasopressin differ at only positions 3 and 8 in
The posterior lobe of the pituitary gland, also called the the peptide chain. Likewise, the sequences of the neurohy-
neurohypophysis and pars nervosa, stores and releases two pophyseal hormones from different vertebrate groups
neurohormones, antidiuretic hormone and oxytocin. These exhibit variations at positions 3 , 4 and 8 (Table 9-4). The
neurohypophyseal hormones are synthesized and packaged sequence of amino acid residues in each pituitary nona-
in the cell bodies of two groups of neurosecretory cells that peptide is, of course, genetically determined. Substitution
comprise the supraoptic and paraventricular nuclei in the of amino acid residues at positions 3 , 4 , and/or 8 during
anterior portion of the hypothalamus (see Figure 9-5). Af- evolution has resulted in several forms of these peptide
ter their synthesis, the hormones are transported within the hormones. The residues that are highly conserved (never
axons of the hypothalamo-hypophyseal tract to nerve ter- undergo substitution) in these hormones are presumably
minals in the neurohypophysis, where they are released into necessary for function; those that are not conserved (po-
a capillary bed. This was the first neurosecretory system sitions 3, 4, and 8) seem to be functionally neutral and
discovered in the vertebrates. probably serve only to place the essential residues in the
Both antidiuretic hormone (ADH),also known as vaso- positions appropriate for the biological activity of these
pressin, and oxytocin are peptides containing nine amino neuropeptides.
acid residues. Both are mildly effective in fostering con- Within their respective neurosecretory cells, the
tractions of the smooth-muscle tissue in arterioles and the neurohypophyseal hormones are covalently linked in a 1:1
uterus (Figure 9-7). In mammals, however, oxytocin is best ratio to cysteine-rich protein molecules termed neuro-
known for stimulating uterine contractions during parturi- physins, which exist in two major types, neurophysin I and
tion and for stimulating release of milk from the mammary neurophysin 11. Oxytocin is associated with type I
gland; in birds, it stimulates motility of the oviduct. The and vasopressin with type 11. Neurophysins have no hor-
foremost function of ADH is to promote water retention in mone activity, even though they are secreted along with the
the kidney, as discussed in a later section. neurohypophyseal hormones. It is conjectured that
Neural stimuli:
1 stress \
Figure 9-7 The two neurohormonesreleasedfrom the mammalIan pos- the neurosecret~onof ant~d~uret~c
hormone (ADH) H~ghplasma-solute
gland function pr~marllyIn reprodualon (oxytoc~n)
ter~orp~tu~tary and reg- concentrat~onand low blood pressure result~ng
from low plasma volume
ulat~onof water balance (ADH) Osmoreceptors In the hypothalamus, st~mulateADH output Oxytocln IS released durlng labor and nurslng
baroreceptors In the aorta, and exterocept~vesensory Input all Influence
TABLE 9-4
Variant forms of neurohypophyseal nonapeptide hormones
Pos~t~ons
of ammo ac~dres~dues*
I
[ Arglnlne vasopressln
! Oxytoc~n Mammals
Some teleosts
t
Ir
lsotocln
I Some elasmobranchs
GnRH
Vasoactive intestinal
peptide
Cholecystokinin
(brain)
Peptide hormones range in length from as few as three amino acid brain-gut hormones.The circles represent individual residues identified
residues to several dozen residues.Of the representative peptide hor- with the three-letter amino acid code (see Table 3-7). TRH = TSH-
mones shown here, the upper three are releasing or release-inhibiting releasing hormone; GnRH = gonadotropin-releas~ng hormone.
hormones produced by hypothalam~cneurons and the lower four are
the hormone-neurophysin molecules are enzymatically sengers, which amplify the signal and mediate rapid,
cleaved upon release into the blood, yielding the neurohy- short-lived responses via various effector proteins.
pophyseal hormone and the neurophysin moiety. Thus neu-
rophysins appear to act as storage proteins, serving to re- The prostaglandins are the exception proving the rule that
tain the hormones in the secretory granules until release. the nature of the receptor, not the hormone, determines the
mode of action. Although lipid-soluble, prostaglandins
bind to cell-surface receptors and have a rapid, short-
CELLULAR MECHANISMS OF lasting effect, similar to that of lipid-insoluble hormones.
HORMONE ACTION Table 9-5 summarizes some properties characteristicof the
As noted already, hormones produce their specific effects major types of lipid-soluble gndlipid-insoluble hormones.
on their target tissues via specialized receptor proteins lo-
cated either inside the cell or on the surface of the cell. Most Lipid-Soluble Hormones and
lipid-soluble (hydrophobic)hormones, such as the steroid Cytoplasmic Receptors
and thyroid hormones, readily penetrate the plasma mem- The lipid-soluble steroid and thyroid hormones are carried
brane and bind to receptors in the cytoplasm of target cells. in the bloodstream complexed with carrier proteins.
In contrast, lipid-insoluble (hydrophilic)hormones cannot Without these carriers, only small amounts of these hor-
penetrate the plasma membrane and therefore bind to re- mones could dissolve in the blood, which is an aqueous so-
ceptors on the cell surface. lution, and they would be taken up completely by the first
The intracellular mechanism of action of a hormone encountered lipids in the circulation. The association of
depends on whether it binds to cytoplasmic or cell-surface lipid-soluble hormones with carriers thus markedly in-
receptors (Figure 9-8): creases the amounts of these hormones that can be carried
in the blood. The binding constants of different carriers
Lipid-soluble hormones bind to cytoplasmic receptors, varies, ensuring adequate rates of hormone delivery to all
forming hormone-receptor complexes that translocate target tissues.
to the nucleus and act directly on the DNA of the cell to Once steroid and thyroid hormones dissociate from
effect long-term changes lasting hours or days. their carrier proteins, they can readily enter and leave neigh-
boring cells by diffusing across the plasma membrane. Ini-
Lipid-insoluble hormones bind to cell-surface receptors, tially, nearly all the hormone appears in the cytoplasm, but
often leading to production of one or more second mes- - in target cells hormone-receptorcomplexes form and move
Llp~d-solublehormone
(Flrst messenger)
Proteln
Llp~d-~nsolublehormone
Second messenger
(Flrst messenger)
Effector proteln
Cell membrane
Figure 9-8 Lipid-soluble and lipid-insoluble hormones differ in their pri- face receptors, triggering an intracellular signaling pathway that may in-
mary intracellular mode of action. (A) Most I~pid-solublehormones move volve a second messenger, which in turn combines with another molecule
through the plasma membrane and combine with ~ntracellularreceptor to produce a metabolically active complex. Although they are lipid sol-
proteins, forming active complexes that act on the genetic machinery to uble, the prostaglandins bind to cell-surface receptors. [Concept by
modulate gene expression. (8) Lipid-insolublehormones bind to cell-sur- M. J. Berridge.]
312 P H Y S I O L O G I C A L PROCESSES
.............................................................................
TABLE 9-5
Comparison of lipid-soluble and lipid-insoluble hormones
Llpld soluble L~pldinsoluble
Source: Adapted from Smith et al., 1983, p. 358. Used with permission of McGraw-Hill.
into the nucleus, so that in time more and more hormone thereby regulating the transcription of specific genes and ul-
appears in the nucleus (Figure 9-9A). Although diffusion of timately production of their encoded proteins. Since the
lipid-soluble hormones in and out of cells is a random lipid-soluble hormones act on the cell's DNA to stimulate
process, these hormones exert effects only on specific target or inhibit production of particular proteins, their effects
cells. Understanding how target and nontarget cells differed persist for hours to days, whereas the effects of lipid-insol-
came from several types of evidence. First, autoradiographic uble hormones usually last only minutes to hours.
studies in the 1960s showed that steroid hormones accu-
mulate In the nuclei oftheir target itlls, but not in thCnuclei
of other cells. This specific aici~n~ulation occurs v c n rapidly , What are the differences in the regulation and
and persists for some time at'tcr the labeled steroid is re- action of lipid-soluble and lipid-insoluble hor-
mo\rc.d from the circulation. These f ndings indic,~ttdthat mones?
rarget cclls must contain steroid hormone-specific receptors,
which are absent from nontarget cells.
Such receptor molecules were found by fractionating
a target tissue incubated with a radiolabeled hormone Lipid-Insoluble Hormones and
and separating the components with different molecular lntracellular Signaling
weights by sucrose density-gradient centrifugation. The re- As noted already, binding of hormone to many cell-surface
ceptor-hormone complex then could be identified by the ra- receptors triggers production of second messengers, which
dioactivity of the labeled hormone. Roger Gorski (1979)and transduce the extracellular hormonal signal into the cell's
associates identified the estradiol receptor in this way, using response. Despite the large number of known hormones
labeled estradiol and rat uterus as the target tissue. They that stimulate second-messenger formation, the most im-
found that the receptor, a protein with a molecular weight of portant second messengers fall into only three distinct
about 200,000, binds estradiol very strongly, and is present groups (Figure 9-10):
in uterine tissues but not in other tissues. Most significantwas
the observation that substances that mimic the hormonal ac- Cyclic nucleotide monophosphates (cNMPs),such as
tion of estradiol in the uterus are all bound by this same re- adenosine 3',S1-cyclicmonophosphate (CAMP)and the
ceptor protein. Other receptor proteins have since been iden- closely related guanosine 3',5'-cyclic monophosphate
tified in target tissues of other lipid-solublehormones. (cGMP)
All the cytoplasmic receptors that bind lipid-soluble
hormones share a highly conserved DNA-binding domain Inositol phospholipids, including inositol trisphosphate
(Figure 9-9B). In the absence of hormone, these receptors (InsP,) and diacylglycerol (DAG)
are bound to an inhibitor protein that blocks the DNA- Ca2+ions
binding domain of the receptor, making it inactive. Binding
of hormone to the receptor causes the inhibitor protein to We'll first examine intracellular signaling systems em-
dissociate, thereby activating the receptor by exposing its ploying each of these second messengers as well as mem-
DNA-blnding site. Once the receptor-hormone complex brane-bound enzyme signaling systems, which don't in-
moves into the nucleus, the DNA-binding domain of the re- volve second messengers. Then we'll see how multiple
ceptor binds specific regulatoty sequences within the DNA, systems can interact to produce complex tissue responses.
HORMONES: REGULATION A N D ACTION 313
...............................................................................
Figure 9-9 Steroid and thyroid hormones move from the bloodstream
into target cells, where they can bind to specific receptors that have a
common domain structure. (A) Mechanism of action of lipid-soluble hor-
mones with cytoplasmic receptors. These hormones diffuse randomly
into and out of nontarget cells without interaction or binding (1). Target
cells for a particular hormone contaln cytoplasmic receptors, consisting
of two subunits, specific for that hormone. Formation of hormone-re-
ceptor complexes preferentiallyretains hormone molecules within target
cells (2). These complexes then accumulate in the nucleus, where they
bind to regulatory elements in the DNA, in most cases stimulating tran-
scription of specific genes into the corresponding messenger RNA (3).
The resulting messenger RNA is then translated into protein by the ribo-
Protein somes (4).(B) Model of common domain structure of cytoplasmic recep-
tors for lipid-soluble hormones. In its inactive state the receptor is bound
to an inhibitor protein that blocks the DNA-binding domain of the re-
ceptor. The binding of hormone to the receptor causes the inhibitor pro-
tein to dissociate, thereby activating the receptor by exposing the DNA-
binding site. [Part A adapted from O'Malley and Schrader, 1976.1
Cyclic nucleotide signaling systems mogenate into fractions and found that the adenylate cy-
The advance of science generally depends on two forms of clase activity disappeared if the cell-membrane fragments
- -
progress. One is the everyday growth of scientific knowl- of the homogenate were removed. It was subsequently dis-
edge by the slow but steady accumulation of data in covered that adenylate cyclase is intimately associated with
thousands of laboratories. Such small-scale incremental a hormone receptor in the membrane. Note that the hor-
progress represents by far the major effort expended by the mones that stimulate adenylate cyclase activity do so with-
entire community of scientists. This type of progress gen- out entering the cell; moreover, neither ATP nor cAMP
erally builds upon the infrequent and often unanticipated readily penetrate the plasma membrane when placed in the
breakthroughs that provide revolutionary new insights or extracellular fluid.
' points of departure. Such breakthroughs open new paths of The discovery of the hormonal stimulation of adenylate
inquiry, which are then explored in detail by the slow step- cyclase provided the first evidence for a link between ex-
by-step mode of progress until at some unexpected time an- tracellular hormones and intracellular regulatory agents
other major breakthrough provides new insight and again and led to the second-messenger hypothesis. The hormone
alters the course of daily investigation. acts on the outer surface of the cell membrane, whereas
An example of such a giant leap occurred in the mid- cAMP is produced enzymatically from ATP at the inner
1950s when the late Earl W. Sutherland and associates dis- surface of the membrane. The hormone conveys its signal
covered the role of cyclic AMP (CAMP)as an intracellular through the surface membrane without having to penetrate
regulatory agent. In his initial studies on CAMP,Sutherland the membrane. Sutherland's findings opened the way for a
noted that the activity of adenylate cyclase, which catalyzes totally new understanding of regulatory processes in many
conversion of ATP to CAMP,is enhanced when certain hor- areas of biochemistry and cell biology. Other researchers
mones are added to cell-free liver homogenates or prepara- subsequently accumulated vast amounts of data that con-
tions of intact cells. He then separated the cell-free ho- firmed the role of CAMPas an intracellular regulatory agent
314 P H Y S I O L O G I C A L PROCESSES
Control of cellular responses Another problem with ducer proteins, GSand G,, are linked to R, and RI, respec-
second-messenger systems is how to reduce or terminate tively. Thus, the activity of adenylate cyclase can be in-
the signal induced by binding of the hormone so that only creased by a stimulatory signal (R, through G,) or reduced
an appropriate response is elicited for an appropriate by an inhibitory signal (R, through GI).Both stimulation
amount of time. Three control mechanisms operate in the and inhibition of adenylate cyclase can occur in the same
CAMPpathway. As we've seen, there are two kinds of re- cell, the end result depending on the intensity of each sig-
ceptors, R, and R,, that bind stimulatory and inhibitory nal. For example, lipid breakdown in fat cells is accelerated
hormones, respectively. Of course, the two types of trans- in response to binding of epinephrine to the stimulatory
316 P H Y S I O L O G I C A L PROCESSES
...............................................................................
Cell exterior I
Regulato
subunit
Proteln k~naseA
I
I
Figure 9-12 Hormone-stimulated regulation of adenylate cyclase (AC)
within the membrane leads to an increase or decrease in cytosolic CAMP
Phosphoprotein
phosphatase
/
t o and removes the regulatory subunit of protein kinase A. The catalytic
subunit, once free ofthe inhibitory regulatory subunit, can phosphorylate
level. Binding of hormones or other ligands t o their stimulatory or in- various intracellular effector proteins, yielding activated phosphoproteins
hibitory receptors (R, and R , respectively) induce binding of GTP to the that mediate cellular responses. In time, CAMP is degraded t o AMP by a
respective transducer proteins, G, and GI.The GTP-activated G proteins phosphodiesterase (PDE), and the phosphorylated effector proteins of-
are then able to either activate or inhibitthe catalytic activity of adenylate ten are dephosphorylated t o their inactive forms. Both of these mecha-
cyclase until the GTP is dephosphorylated enzymatically t o guanosine nisms reduce or terminate the effects of the external signal. [Adapted
diphosphate (GDP) and the effect on the cyclase ceases. Activated from Berridge, 1985.1
adenylate cyclase catalyzes the conversion of ATP t o CAMP, which binds
P-adrenoreceptors, whereas it is decreased in response to A second control mechanism involves the intracellular
binding of either epinephrine or adenosine to the inhibitory level of CAMP,which depends not only on its rate of syn-
a-adrenoreceptors or adenosine receptors. thesis from ATP, but also on its rate of degradation to
adenosine 5'-phosphate (AMP):
chapter?
ATP - - 1
CAMP
2
AMP
2 ATP 3
-
f
cyclic AMP Regulatory
3
Protein
f - subunit
b :
1
kinase A
Protein CAMP
Inhibited
kinase A
Active
ATP
Phosphorylase
kinas; 1
lnactive Phosphorylase
kinase-PO,
Cap+ * Active
ATP
Phosphorylase b
Inactive 1
Phosphorylase a-PO,
Active
Glucose-6-PO, -+Glucose
Glycolysis and
Krebs cycle
Figure 9-13 Epinephrine and glucagon stimulate breakdown of glyco- which several enzymes are converted from an inactive to active form. As
gen to glucose (glycogenolysis) in muscle and liver, respectively. Hor- a result of this enzyme cascade the original signal IS greatly amplified. See
mone binding to P-adrenoreceptorstriggers a sequence of reactions in text for discussion. [Adapted from Goldberg, 1975.1
enzyme, phosphorylase kinase (step 4), thereby activating The CAMP-dependentprotein kinase A that stimulates
it. Phosphorylase kinase-PO, in turn catalyzes phosphory- formation of phosphorylase a also acts in an indirect way
lation of phosphorylase b to form the active form, called to inhibit glycogen synthetase, the enzyme that catalyzes
phospho;rylase a (step 5). It is the latter enzyme that the polymerization of glucose into glycogen. Thus a
cleaves glycogen with addition of PO-: to form glucose 1- hormone-stimulated increase in intracellular cAMP stim-
phosphate (step 6). In cells, glucose 1-phosphate is read- ulates breakdown of glycogen and inhibits its synthesis.
ily converted to glucose 6-phosphate, which enters the This synergistic effect is important, for it keeps the rise in
glycolytic pathway or is dephosphorylated to glucose, glucose from driving by mass action the resynthesis of
which is transported across the plasma membrane into the glycogen from glucose. Conversely, a decrease in CAMPin-
bloodstream. hibits glycogen breakdown and stimulates glycogen syn-
thesis. This example illustrates that multiple CAMP-medi- tance as important second messengers in transducing many
ated effects can occur simultaneously within a single cell. hormonal and other extracellular signals into a wide vari-
Spotlight 9-2 describes the amplification of the hormone ety of cellular responses.
signal during glucose mobilization.
Figure 9-14 Binding of hormones by some G protein-linked receptors activation of guanylate cyclase by PLC (dashed pathway) is not fully es-
induces formation of the phospholipid-derived second messengers di- 'tablished. Note that Ca2+mobilized from intracellularstoresmay activate
acylglycerol (DAG) and inositol trisphosphate (InsP,). (A) Genericscheme troponin C (TnC), form a complex with calmodulin (CaM) that activates
of inositol phospholipid pathway, which is nearly identical with that of the Ca2+/calmodulin-dependent kinase (Ca2+/CaMkinase), promote activa-
CAMPpathway (see Figure 9-1IA). (B) Condensed outline of the inositol tion of protein kinase C, or increase cGMP production by stimulating
phospholipid second-messenger system. The amplifier enzyme in this membrane-bound guanylate cyclase.
pathway is phosphoinositide-specific phospholipase C (PLC). The direct
and activates troponin C (TnC)and calmodulin (CaM),as act as an effector protein, or bind to and activate a number
well as a number of other regulator and effector molecules of enzymes and other effector proteins, of which the most
(see Figure 9-14).As we'll discuss in Chapter 10, Ca2+/TnC studied is Ca2+/calmodulinkinase. These proteins induce
stimulatesmuscle contraction directly. Ca2+/calmodulinmay various cellular responses through different mechanisms.
HORMONES: REGULATION A N D ACTION 321
...............................................................................
II - Protein (inactive)
+ ADP
Figure 9-15 In the inositol phospholipid system, one second messenger of the membrane-bound protein kinase C; this activation also requires
acts in the membrane and the other in the cytosol. Hormone binding in- Ca2+ and phosphatidylserine (PS), another membrane phospholipid.
ducesformation of the coupled GTP-activated G protein, which activates InsP3promotes the liberation of Ca2+from intracellular storage depots
phosphoinositide-specific phospholipaseC (PLC).PLCthen catalyzes hy- like the endoplasmic reticulum. The free Ca2+has numerous regulatory
drolysis of membrane phosphatidylinositol4.5-biphosphate (PIP,) into di- functions, including stimulation of Ca2+/calmodulinkinase (Ca2+/CaMki-
acylglycerol (DAG),which remains in the membrane, and inositol trispho- nase). [Adapted from Berridge, 1985.1
sphate (InsP,), which diffuses into the cytosol. DAG promotes activation
The actions of DAG, the other second messenger in the cytosol. Thus, the activation of protein kinase C requires
IP system, occur in the plasma membrane, in which DAG two intracellular messengers, DAG and Ca2+, both of
molecules can move laterally by diffusion. DAG has two which can be induced by the same extracellular signal.
potential signaling roles. First, it can be cleaved to release Among the tissue-specific responses induced by hor-
arachidonic acid, a precursor in synthesis of the pros- mones via the inositol phospholipid pathway are the fol-
taglandins and other biologically active eicosanoids. Sec- lowing (Berridge, 1985):
ond, and more important, DAG activates membrane-
bound protein kinase C by a mechanism analogous to Breakdown of liver glycogen stimulated by vasopressin
activation of protein kinase A by CAMP.Although protein DNA synthesis in fibroblasts stimulated by growth
kinase C occurs in both the cytosol and in the inner portion factors
of the cell membrane, it can only be activated when asso-
ciated with the membrane. The activation of protein kinase Secretion of prolactin from anterior pituitary gland
C by DAG depends on Ca2+and phosphatidylserine (PS), stimulated by thyrotropin-releasing hormone
another phospholipid constituent of the membrane. Bind-
ing of DAG and PS to protein kinase C, located in the cy- Ca2+ signaling systems
toplasmic half of the membrane, increases the affinity of the The concentration of free Ca2+in the cytosol can be in-
enzyme for Ca2+;as a result, protein kinase C can be acti- creased in two ways: (1)release of Ca2+from intracellular
vated at the usual low concentrationsof Ca2+present in the calcium stores such as the endoplasmic reticulum (called
322 PHYSIOLOGICAL P R O C E S S E S
...................................
S P O T L I G H T 9-2 active) molecule into an active enzyme. The result is a progres-
sive amplification through four steps. In addition, each molecule
of the final effector in this pathway, phosphorylase a, can itself
AMPLIFICATION BY
convert many glycogen molecules into glucose I-phosphate. If
ENZYME CASCADES it is assumed, conservatively, that each activated enzyme mole-
cule catalyzes the activation of 100 molecules in the next step,
The cAMPsignaling pathway stimulated by binding of epineph- then the five amplification steps would produce an overall
rine or glucagon and leading t o glycogen breakdown exempli- amplification of 10lO.That is, the interaction of one molecule
fies the amplification of hormone action by enzyme cascades. of glucagon or epinephrine with its membrane receptor in
The pathway between hormone binding and the breakdown of liver or muscle cells can result in the mobilization of about
glycogen is rather complex (see Figure 9-13). It would be simpler, 10,000,000,000 or more molecules of glucose.
of course, for CAMPto activate the final enzyme in the sequence The basal intracellular concentration of CAMP, the key t o
directly, thereby saving several steps in this multistep sequence. transmitting the signal from the membrane to the cytosol, isvery
O n the other hand, the large number of steps makes sense if we low (10-l2 t o M).Thus the hormone-induced production of
consider the need for amplification-that is, for producing a only a small absolute number of cAMP molecules will represent
large effect in response t o a few hormone molecules. a large percentage increase in cAMP concentration. Binding of
The diagram shows the steps in this pathway at which bio- just a few hormone molecules therefore can produce significant
chemical amplification occurs. Steps 1,2,4, and 5 involve an ac- changes in cAMP levels and ultimately mobilize large amounts
tivating reaction that converts a catalytically inactive (or weakly of glucose from glycogen in a short time.
(1) Ampllficatlon
/I'
through G prote~n
cAMP
(3)
Protein kinase A
(4) Amplification / 1
e e .4 Phosphorylase kinase-PO,
Amplification
Phosphorylase a
(6) Amplification
f
/ /
77 1
Glycogen
0 0 0 0 OGlucosel-phosphate
Enzyme cascades greatly amplify hormone action. In the pathway of hor- cading of amplifying steps in the form of a series of enzyme-activating re-
rnonal stimulation of glycogenolysis, biochemical amplificationoccurs at actions can explain the extremely high potencies of many hormones. See
several steps, so binding of one molecule of ep~nephrineor glucagon can Figure 9-13 for reaction sequence. [Adapted from H. D. Lodish et al.,
lead to production of 10'' molecules of glucose I-phosphate. Such a cas- 1995.1
sarcoplasmic reticulum in muscle) and (2)influx of Ca2+ stimulated by InsP,, phosphorylation of the Ca2+channel
from the cell exterior through Ca2+channels in the plasma by CAMP-dependentkinase, electrical stimulation, or re-
membrane. As described in the previous section, InsP, stim- ceptor activation itself, which we will discuss later.
ulates release of calcium from intracellular calcium stores. In recent decades it has become clear that Ca2+plays
The entry of Ca2+ from the cell exterior, through Ca2+ an important and ubiquitous role as both an intracellular
channels in the plasma membrane, has been shown to be regulatory agent and a messenger linking external signals
to cellular responses. Two important characteristics of cells Ca2+in cells. The cell has two primary mechanisms for re-
permit Ca2+to function effectively in cellular regulation moving excess Ca2+from the cytosol, thereby keeping the
and signaling: (1)the ability of cells to increase and de- free Ca2+level low: primary and secondary active transport
crease the intracellular Ca2+level over a wide concentra- of Ca2+ across the plasma membrane to the exterior (see
tion range and (2)the presence within cells of numerous Chapter 4) and movement of Ca2+ions into the endoplas-
protelns-whose activity is modulated by the binding of mic reticulum via a Ca2+pump in the reticulum membrane.
Ca2+.We'll first discuss these aspects of calcium's role in Two additional mechanisms help keep the intracellular
the cell and then look at how Ca2+functions as a second Ca2+level from transiently becoming too high. First, vari-
messenger. ous cytosolic proteins bind Ca2+when the Ca2+ level in-
creases and release Ca2+when the level decreases. In effect,
Modulation of intracellular Ca2+ concentration Most these proteins "buffer" the Ca2+ concentration, limiting
Ca2- ions that enter the cell from the exterior are rapidly perturbations in free Ca2+ levels, just as p H buffers limit
bound to anionic sites on protein molecules in the cytosol; perturbations in free H+ levels. Second, when the cytoso-
only a small percentage of the ions remain ionized and lic Ca2+level becomes abnormally high, the mitochondria
free to diffuse. As a result, although the total Ca2+content may import Ca2+in exchange for H+.
of most cells is about 1 mM M), the concentration Certain technical advances have been essential in
of free, ionized Ca2+in the cytosol is maintained at extra- studying the physiological effects of changes in the intra-
ordinarily low levels, usually below lo-' M. (Note that cellular Ca2+concentrations. One of these advances was
unless indicated otherwise, references to intracellular lev- the discovery in 1963 of the jellyfish protein aequorin,
els of Ca2+ and other ions refer to free, unbound ions.) which emits light when it complexes with Ca2+. Since
The advantage of this very low intracellular Ca2+concen- light can be measured with very sensitive instruments, in-
tration is quite simple: influx of quite small amounts of jection of aequorin into cells has provided a means of
Ca2+from the extracellular space produces a very large in- detecting minute changes in the intracellular free Ca2+
crease in the concentration of free Ca2+ in the cytosol. level. More recently, calcium-sensitive dyes (e.g., arsenazo
This concept is illustrated by comparing the relative 111) and calcium-sensitive fluorescent molecules (e.g.,
changes in the intracellular concentration of Ca2+ and quin-2 and fura-2) have opened up new possibilities for
Na+ that result from the entry of equal quantities of these sensitive optical measurement of Ca2+levels within single
two ion species in response to a transient increase in the living cells.
permeability of the plasma membrane to both ions (Fig-
ure 9-16). Likewise, release of small amounts of Ca2+ Ca2+-bindingproteins The other important feature typical
from intracellular calcium stores causes a relatively large of Ca2+-mediatedintracellular control and signaling is the
increase in the concentration of free Ca2+ in the cytosol. presence of multiple Ca2+-bindingsites in certain enzymes
Thus, the cell maintains the cytosolic Ca2+level extremely and various regulatory proteins. These specialized binding
low, permitting it to increase as much as tenfold as a con- sites have a very high affinity for Ca2+,allowing tight bind-
sequence of Ca2+flow into the cell or out of the intracel- ing of the cation at very low concentrations of free Ca2+.
lular calcium stores. The Ca2+-bindingsites in all these proteins consist of acidic
Since the extracellular Ca2+concentration is typically amino acid residues, which are negatively charged and rich
about M, the electrochemical gradient favors entry of in oxygen atoms. The oxygen atoms, carrying full or
10-8~
+
1 influx
1o-2 M
+
1 influx
1 0 - 6 ~- 1.0001 X 10-z M
-
[Na+],,,,, = 1.0001 X lo-' = 1 x initial [Na+]
[Na+llnlt A[Na+I [Na+I11na1 [Na+Ilnlt lo-'
Figure 9-16 The ~ntracellularconcentrat~on of free Ca2+IS elevated many fold by a transient ~nflux,A[Ca2+],equivalent to a M Increment (B)
fold by lnflux of small amounts of Ca2+ (A) In this example, the low inl- ,,
Slnce [Na+] 1s already 10 M, a 10 M increment,A[Nai], produces vr
tlal lntracellular concentratlon of Ca2+,[Ca2+],,,,, of lo-* M IS rased 100- tually no change In the lntracellular Na+ concentratlon
Binding site Figure 9-17 Calmodulin, a cytosolic protein with four Ca2+-bindingsites,
forms the Ca2+/calmodulincomplex, an important intracellular regulator.
ca2+ (A) Amino acid sequence of the Ca2+-bindingsite at the C-terminus of
/
calmodulin. Each binding site contains aspartate, glutamate, and as-
paragine residues, shown in orange, whoseside chains form ionic bonds
with a Ca2+ion, forming a loop in the backbone. Other binding sites also
contain threonine and serine residues, whose side chain oxygen atoms
also associate with the Ca2+ion. (B) Model of the calmodulin molecule
with four bound Ca2+ions (blue spheres). (C) Diagram illustrating Ca2+-
induced conformational change in calmodulin. Calmodulin undergoes a
conformation change when all four Ca2+-bindingsltes are occupied. The
resulting Ca2+/calmodulin complex can bind to numerous target pro-
teins, modulating their activity. [Part B courtesy Y. S. Babu and W. J. Cook;
parts A and C adapted from Lodish et al., 1995.1
partial negative charges, occur in a loop of the peptide eukaryotic tissue examined thus far. It functions as a mul-
chain, so that six to eight oxygen atoms form a cavity of tipurpose intracellular regulatory protein, mediating most
just the right size to harbor the positively charged calcium Ca2+-regulatedprocesses. The single polypeptide chain of
ion (Figure 9-17A).In fact, about 70% of the entire amino calmodulin, consisting of 148 amino acid residues, con-
acid sequences of various Ca2+-bindingregulatory proteins tains four Ca2+-bindingsites (Figure 9-17B). Binding of
are homologous. Ca2+to all four sites produces a Ca2+/calmodulincomplex
Binding of Ca2+to these proteins generally leads to that can bind to and activate numerous enzymes and ef-
a conformational change in the molecule. This change fector proteins (Figure9-17C). For example, Ca2+/calmod-
in conformation can produce an allosteric effect that al- ulin binds to the regulatory subunit of Ca2+/calmodulinki-
ters the properties of the molecule. For example, binding nase. Once freed of its regulatory subunit, the catalytic
of Ca2+to troponin C, which is found only in striated subunit of Ca2+/calmodulinkinase can phosphorylate ser-
muscle, causes a conformational change in the molecule ine and threonine residues on various effector proteins,
that initiates a series of steps leading to contraction and which induce cellular responses (see Figure 9-14). Other
is important in regulating contraction of vertebrate enzymes and cellular processes regulated by Ca2+/calmod-
striated muscle. We'll discuss troponin C, the first Ca2+- ulin are shown in Figure 9-18. Note that Ca2+/calmodulin
binding regulatory protein to be discovered, in detail in activates myosin light-chain kinase, a protein that regu-
Chapter 10. lates contraction in vertebrate smooth muscle; this func-
Calmodulin, a Ca2+-bindingprotein closely related to tion is somewhat analogous to that of troponin C in ver-
troponin C, is present in relatively large amounts in every tebrate striated muscle.
HORMONES: REGULATION A N D ACTION 325
...............................................................................
Adenylate
Myosin llght 'yclase
cham kinase Phosphodiesterase
f l Phosphol~pases
-
Phosphorylase k~nase
K
Guanylate cyclase- Calcium-
Calmodulin Ca2+-ATPase
Ca2+-dependent / \ disassembly?
Microtubule
protein kinase I, II
Neurotransmitter Membrane
release? phosphorylation
Others
Figure 9-18 Calcium/calmodulin regulates many processes or enzymes which catalyze formation of the nucleotide second messengers.
within cells. Among these are adenylate cyclase and guanylate cyclase, [Adaptedfrom Cheung, 1979.1
Second-messenger role of Ca2+ Earlier we learned that pathways. Since the receptor guanylate cyclase has a ligand-
Ca2+acts as a third messenger in the inositol phospholipid binding domain, it can be activated directly by hormone
system. Stimulation of other receptor systems leads to an binding. The cGMP produced by activation of this recep-
influx of Ca2+,which then can act as a second (and only) tor can function as a second messenger to mediate cellular
messenger, as illustrated in Figure 9-19. Various signals can responses as in other pathways.
activate Ca2+second-messenger pathways. For example,
activation of a-adrenoreceptors by epinephrine in the Second-messenger networks
mammalian liver and salivary gland stimulates Ca2+influx It is important to note that a single hormone can trigger
through opening of Ca2+ channels in the plasma mem- several second-messenger systems by activating different
brane, whereas membrane depolarization causes opening types of receptors, even in the same cell. Binding of epi-
of Ca2+channels in muscle. nephrine to a - and p-adrenoreceptors in the mammalian
salivary gland is an example of a divergent pathway in
Membrane-enzyme signaling systems which the two second messengers-namely, intracellular
Some cell-surface receptors seem to signal the cell directly free Ca2+and CAMP,respectively-mediate different cell
through their intrinsic enzyme activity. Such receptors have responses (Figure 9-21A). In mammalian liver, however,
a ligand-binding domain on the extracellular surface of the binding of epinephrine to a- and P-adrenoreceptors leads
plasma membrane and a catalytic domain on the intracel- to the same cell response, an example of a convergent path-
lular surface. Binding of an external signal to this type of re- way (Figure 9-21B). In this case, both second messengers,
ceptor triggers a conformational change that causes the cat- Ca2+and CAMP,activate phosphorylase kinase, which in
alytic domain to become activated. The activated catalytic turn stimulates glycogenolysis as discussed earlier.
domain, in turn, induces further intracellular changes that A more complicated example of second-messenger net-
result in the cellular responses. works involves serotonin (5-hydroxytryptamine, 5-HT),a
To date, cell-surface receptors with intrinsic protein ki- lipid-insoluble amine that functions as both a neurotrans-
nase or guanylate cyclase activity have been identified. The mitter and an endocrine hormone regulating gastric secre-
best-studied of these in animal cells are receptor tyrosine tion and smooth muscle contraction in blood vessels. As
kinases (RTKs), which are known to bind insulin and shown in Figure 9-22, serotonin binds to several receptor
a number of growth factors including platelet-derived subtypes, which are linked to various second-messenger
growth factor (PDGF).When activated by external signal pathways or ion channels; some of these converge, while
binding, RTKs transfer the phosphate group from ATP to others diverge. Like other lipid-insoluble hormones, sero-
the hydroxyl group on a tyrosine residue of selected pro- tonin binds to cell-surface receptors; however, its mode
teins in the cytosol (Figure 9-20A). In all cases studied, of action, which ultimately affects gene transcription, dif-
RTKs also phosphorylate themselves when activated; this fers from that of most lipid-insoluble hormones (see
autoph~sphor~lation enhances the activity of the kinase- Figure 9-8B).
an example of positive feedback regulation. Atrial natri- As we saw earlier, activation of different receptors may
uretic peptide (ANP)has been shown to activate a receptor stimulate or inhibit the same signaling system; for exam-
guanylate cyclase (Figure 9-20B).A glance back at Figures ple, norepinephrine stimulates, but neuropeptide Y in-
9-14 and 9-19 shows that membrane-bound guanylate cy- hibits, the inositol phospholipid system. Another possible
clase can be activated by Ca2+generated in other signaling pattern is a single receptor capable of coupling to two
326 PHYSIOLOGICAL PROCESSES
...............................................................................
Chem~cal
troponin C,
and other
i
Ca2+
Ca-dependent
protelns
Prote~nk~naseG
ATP + ATP +
I
I-
Phosphoprotein (active)
protein (inactive)
+ ADP
protein (inactive)
Figure 9-19 St~mulat~on of receptors that function as calc~urn-select~ve cytosol The resulting local Increase In cytosol~cfreeCa2+from a rest~ng
Ion channels causes an ~nfluxof Ca2+,wh~chacts as a second messenger level of M to M can act~vateseveral d~fferentlntra-
E~thermembrane depolar~zat~on or b~ndingof a chem~calmessenger cellular s~gnal~ng
pathways, lead~ngto varlous cell responses CaM =
(e g , extracellular hormone) can "open" an ion channel, permlttlng Ca2+ calmodul~n
to move through the channel down ~ t selectrochem~calgradient ~ n t the
o
ATP ADP /
Substrate protein Phosphorylated
substrate protein
,
A Divergent pathway B Convergent pathway
Epinephrine Epinephrine
/ \ \
Adenylate cyclase
I Fluid secretion
DAmylase release
Mammalian liver
Figure 9-21 Asingle hormone may bind to different receptors, initiating messengers both induce activation of phosphorylasekinase, which cat-
convergent and/or divergent s~gnalingpathways. Binding of epinephrine alyzes the breakdown of glycogen to glucose (glycogenolysis)(see Fig-
to a- and P-adrenoreceptors leads to increases in intracellular Ca2+and ure 9-13). Thus, binding of the same hormone to different receptors trig-
CAMP,respectively. In the mammalian salivary gland (A) these two sec- gers convergent pathways leading to the same end response.There is
ond messengers mediate divergent pathways, leading to different, in- growlng evidence that epinephrine is not unlque in having mult~plere-
dependent end effects-fluid secretion and amylase secretion by se- ceptors in the same animal, or even in the same cell.
cretory cells in the gland. In mammalian liver (B), these two second
Figure 9-22 Serotonin binds to multiple receptors, which are linked to represent subclasses of the serotonin-receptor family (dro = Drosophila).
convergent and divergent second-messenger pathways. Binding of sero- GI = inhibitory G proteins; G, = stimulatory G proteins; G, = pertussis
tonin, also known as 5-hydroxytryptamine(5HT), to some receptors leads toxin-insensitive G proteins; AC = adenylate cyclase; PLC = phospho-
to production of CAMP,diacylglycerol (DAG), or inos~toltrisphosphate lipase C; ER = endoplasmic reticulum; PKA = protein kinase A; PKC =
(InsP,), which all can mediate the same cellular responses in cells of dif- protein kinase C; Ca2+/CaMkinase = Ca2+/calmodulin-dependentpro-
ferent tissues, or even in the same cells. The various receptors illustrated tein kinase. [Adapted from Saudou and Hen, 1994.1
PHYSIOLOGICAL EFFECTS marizes the properties of the major metabolic and devel-
OF HORMONES opmental hormones.
As noted earlier, most hormones produce tissue-specific Glucocorticoids and catecholamines
physiological effects. That is, a given hormone generally The adrenal gland, which is situated close to the kidney, is
induces responses only in selected tissues, and may in- actually composed of two developmentally and function-
duce different responses in different tissues. This speci- ally unrelated glandular tissues: an outer cortex, derived
ficity in hormonal action depends partly on the restricted from nonneural tissue, and an inner medulla, derived from
distribution of the components of hormone-triggered the neural crest (see Figure 8-12). As described in Chapter
signaling pathways (especially receptors) and partly 8, the adrenal medulla synthesizes and secretes the cate-
on the preferential expression of effector proteins in cholamines epinephrine and norepinephrine, which can
different tissues. In the following sections, we'll examine bind to a- and P-adrenoreceptors. Stimulation of a-adreno-
the physiological effects of four major categories of receptors may cause a decrease in CAMPvia a coupled in-
hormones. hibitory G protein (G,);may trigger the inositol phospho-
lipid pathway, leading to release of Ca2+from intracellular
Metabolic and Developmental Hormones stores; or may activate an associated Ca2+channel, leading
Several different hormones regulate metabolism and vari- to an influx of extracellular Ca2+into the cell. Stimulation
ous developmental processes. Produced in various en- of p-adrenoreceptors usually is coupled, via a stimulatory
docrine tissues, these hormones have diverse structures G protein (G-),to an increase in CAMP.The catecholamines
(e.g., steroids, catecholamines, peptides). Table 9-7 sum- affect contraction of smooth muscle, induce vasoconstric-
TABLE 9-7
Metabolic and developmental hormones
tion, and stimulate glycolysis and lipolysis. Their physio- the ACTH-secreting cells of the anterior pituitary gland
logical effects are discussed in Chapter 8 and summarized (Figure 9-24). The basal level of glucocorticoid secretion
in Table 8-2. also undergoes a diurnal rhythm resulting from cyclic
In this chapter, we focus on the hormones produced by variation in CRH secretion, which appears to be influ-
cells of the adrenal cortex. When stimulated by ACTH, the enced by an endogenous biological clock. Basal glucocor-
adrenal cortex synthesizes and secretes a family of steroids ticoid levels in humans are maximal during the early
derived from cholesterol (Figure 9-23). These hormones fall hours of the morning prior to waking. This is adaptively
into three functional categories: useful because of the energy-mobilizing actions of these
hormones. In addition to such endogenous regulation of
Reproductive hormones secretion, the adrenal cortex is stimulated to secrete glu-
cocorticoids in response to stress of various types (includ-
Mineralocorticoids, which regulate kidney function
ing starvation). Stress, acting through the nervous system,
Glucocorticoids, which have widespread actions, in- causes an elevation in ACTH and hence stimulation of
cluding mobilization of amino acids and glucose and the adrenal cortex.
anti-inflammatory actions
Progesterone
Aldosterone CH,OH Cortisol
CH,OH I
C=O CH,OH
'-A
-/
0
0
Androstenedione
0
Testosterone
YH
Estradiol
OH
Figure 9-23 Cholesterol is the precursor for three major classes of hor- aldosterone and cort~solare the prlmary ones, respect~vely,In mammals
mones: mineralocorticoids, glucocort~coids,and reproductive hormones. The adrenal cortex 1s the prlmary s~tefor secretion of these hormones
Modificationsto the cholesterol structure, shown in boldface, yield a large The reproductive hormones (progesterone,testosterone, estrone, estra-
number of related steroid hormones and intermediates. (Some interme- d ~ o lare
) secreted mostly from the gonads, although they also are se-
diates have been omitted in the synthetic pathway shown here.) Several creted from the adrenal cortex
steroid hormones have mineralocorticoid or glucocorticoid activity, but
into glycogen, which is stored in the liver and muscle. Most maintain adequate blood glucose to sustain energy pro-
of the newly produced glucose, however, is released into the duction in critical tissues such as the brain. The glucocor-
circulation, causing a rise in blood glucose levels, but the ticoids also stimulate mobilization of fatty acids from stores
glucocorticoids also act to reduce uptake of glucose into pe- of fat in adipose tissue. These can be used as substrates for
ripheral tissues such as muscle. At the same time, the up- gluconeogenesis in the liver or metabolized directly in mus-
take of amino acids by muscle tissues is decreased by glu- cle to provide energy for contraction. All these actions thus
cocorticoids, and amino acids are released from muscle increase the availability of quick energy to muscle and ner-
cells into the circulation. This release increases the quantity vous tissue. The glucocorticoids have numerous other ac-
of amino acids available for deamination and conversion tions including stimulation of gastric secretion and inhibi-
into glucose in the liver under glucocorticoid stimulation. tion of immune responses.
This mechanism is especially important during starvation, As discussed earlier, the glucocorticoids, like other
the end result being the degradation of tissue proteins to lipid-soluble steroid hormones, bind to specdic receptors in
Stress; neural input
Circadian rhythm
Figure 9-24 Secretion of glucocorticoids, and hence their effects on tar- cretion of glucocorticoids by the adrenal cortex. These steroids produce
get tissues, is regulated by neural stimuli and negative feedback. Neural an increase in blood glucose and liver glycogen by stimulating conver-
stimuli induce release of corticotropin-releasinghormone (CRH)from hy- sion of amino acids and fats to glucose. Negative feedback by the glu-
pothalamic neurosecretory cells. Subsequent release of adrenocorti- cocorticoids to both the pituitary and the hypothalamusmay limit ACTH
cotropin hormone (ACTH)from the anterior pituitary gland stimulates se- release.
the cytosol, forming hormone-receptor complexes that en- regulation, however, is the stimulation of the hypothalamus
ter the nucleus and regulate the transcription of specific by stress; a low skin temperature, for example, will stimu-
genes (see Figures 9-8 and 9-9). late the release of hypothalamic TRH.
The thyroid hormones act on the liver, kidney, heart,
Thyroid hormones nervous system, and skeletal muscle, sensitizing these tis-
The follicles of the thyroid tissue are stimulated by thyroid- sues to epinephrine and stimulating cellular respiration,
stimulating hormone (TSH) to synthesize and release two oxygen consumption, and metabolic rate. The acceleration
major thyroid hormones-3,5,3'-triiodothyronine (T3) of metabolism stimulated by thyroid hormones leads to a
and thyroxine (T4)-from two iodinated tyrosine precur- rise in heat production. This is of major importance in the
sors (Figure 9-25). Iodine is actively accumulated by the thermoregulation of many vertebrates (see Chapter 16).
thyroid tissue from the blood. The secretion of thyroid hor- The thyroid hormones also significantly affect the de-
mones is regulated by negative feedback of these hormones velopment and maturation of various mammalian verte-
on the hypothalamic neurons that secrete TSH-releasing brate groups. The developmental effects of thyroid hor-
hormone (TRH) and on the TSH-secreting cells of the an- mones occur only in the presence of growth hormone
terior pituitary gland (Figure 9-26). Superimposed on this (GH),and vice versa. The synergistic actions of the thyroid
P H Y S I O L O G I C A L PROCESSES
.........................................................................
Tyrosine
CH2-CH-COOH
I
NHz
Triiodothyronine (T,)
HO CH2-CH-COOH
I
NH2
Tetraiodothyronine
(Thyroxine, T,)
Figure 9-25 The thyroid hormones are produced from iodinated deriv- hormone are linked by an ether bond. T, is also produced by removal of
atives of the amino acid tyros~ne.Condensation of the tyrosine derivatives one iodide from thyroxine.
yields 3,5,3'-triiodothyronine (T,) and thyroxine (T,); the two rings in each
hormones and growth hormone promote protein synthesis Insulin and glucagon
during development. Hypothyroidism resulting from the Insulin is secreted by the beta cells of the pancreatic islets of
lack of dietary iodine during early stages of development in Langerhans, small patches of endocrine tissue scattered
fish, birds, and mammals results in a deficiency disease throughout the exocrine tissue of the pancreas. High blood
(called cretinism in humans) in which somatic, neural, and glucose acts as the major stimulus to the pancreatic beta
sexual development are severely retarded, the metabolic cells to secrete insulin (Figure 9-27).The release of insulin
rate is reduced to as little as about half the normal rate, and is also stimulated by glucagon, growth hormone, gastric in-
resistance to infection is reduced. Inadequate production of hibitory peptide (GIP, also known as glucose-dependent
thyroid hormones, leads to excessive production of TSH insulin-releasing peptide), epinephrine, and elevated levels
due to decreased negative feedback to the hypothalamus of amino acids.
and the anterior pituitary gland. The resulting overstimu- Insulin has important effects on carbohydrate, fat, and
lation of the thyroid gland by TSH causes hypertrophy of protein metabolism. With regard to carbohydrate metabo-
the gland (goiter). Raising the dietary iodine level increases lism, insulin has two major actions: increasing the rate
thyroid hormone production, thereby establishing normal of uptake of glucose into cells of liver, muscle, and adipose
feedback control on TSH production. Thus, the incidence tissue and stimulating glycogenesis (polymerization of glu-
of both cretinism and goiter have been reduced in areas cose to glycogen). As for lipid metabolism, insulin stimu-
where table salt is routinely "iodized" and the population lates lipogenesis in liver and adipose tissue. In protein
is no longer dependent on natural trace amounts of iodine metabolism, insulin stimulates the uptake of amino acids
in food (mainly in seafood). into liver and muscles and the incorporation of amino acids
The thyroid hormones, like the steroid hormones, are into protein.
lipid soluble and bind to specific receptors within the cy- Diabetes mellitus in humans, which occurs in two ma-
tosol. Both types of hormones exert their effects by regu- jor forms, is characterized by an absolute or relative defi-
lating the transcription of specific genes and ultimately the ciency of insulin. Type I diabetes mellitus is associated with
production of the proteins encoded by these genes. For this a loss of pancreatic beta-cell mass, which leads to dimin-
reason, the effects of these hormones develop slowly. For ished or decreased insulin production and secretion (i.e.,
example, it may take up to 48 hours after a rise in blood absolute insulin deficiency). Type I1 diabetes mellitus, on
levels of thyroid hormones before their effects are seen. the other hand, is associated with defective insulin recep-
HORMONES: REGULATION A N D ACTION 333
...............................................................................
Neural stimuli from
temperature-regulating centers
Hypothalamus
Thyroid hormones
+
+ ' +
MUSCLE HEART LIVER KIDNEY
Figure 9-26 The thyroid hormones, which regulate metabolism in vari- cause increased metabolism in skeletal and cardiac muscle, liver, and kid-
ous tissues, are regulated by neural stimuli and negativefeedback. A low ney and hence lead to the metabolic generation of heat. Feedback inhi-
skin temperature and stress stimulatesTSH-releasinghormone (TRH) re- bition by thyroid hormones apparently occurs at the levels of both the
lease from hypothalamic neurosecretory cells; TRH then stimulates se- anterior pituitary gland and the hypothalamus.The follicle shown super-
cretion of thyroid-stimulating hormone (TSH)from the anterior pituitary imposed on the thyroid gland is drawn at a disproportionately large scale.
gland. The thyroid responds hy secreting the thyroid hormones, which
/
glucose
/ I p-cel
(insulin)
Glucaaon Insulin
Figure 9-27 The pancreatic hormones insulin and glucagon play a ma- cells to secrete insulin, which stimulates glucose uptake in all tissues.
jor role in regulating blood glucose levels. High levels of blood glucose Glucagon, secreted by pancreatic a-cells, exerts an action that is antag-
and glucagon andlor gastrointestinalhormones signaling food ingestion onistic to that of insulin in the liver, where it stimulates glycogenolysis and
(e.g.,gastrointestinal inhibitorypeptide,G1P)stimulatethe pancreaticp- glucose release. Insulin has several other effects.
cyclase and activate cAMP phosphodiesterase. This dual releasing hormone (GRH) and GH-inhibiting hormone
action has the effect of lowering intracellular cAMP levels. (GIH),otherwise known as somatostatin (seeTable 9-2).In
Glucagon is secreted by the alpha cells of the pancreatic addition, the release of GRH and GIH is regulated by such
islets in response to hypoglycemia (low levels of blood glu- factors as blood glucose levels (Figure 9-28). Reduced glu-
cose). This hormone has the opposite effects of insulin, cose levels, for example, indirectly stimulate release of
stimulating glycogenolysis in the liver; it also stimulates growth hormone by increasing the secretion of GRH.
lipolysis, providing lipids for gluconeogenesis (see Figure Growth hormone exerts both metabolic and develop-
9-27). The antagonistic actions of insulin and glucagon are mental effects. Many of its diverse metabolic effects are op-
important in maintaining an appropriate blood glucose posite to those of insulin. For example, growth hormone in-
level, so that adequate glucose is available for all tissues. duces the mobilization of stored fat for energy metabolism,
Like epinephrine, which also promotes breakdown of whereas insulin induces the breakdown of stored fat. The
glycogen, glucagon binds to receptors linked to the CAMP fatty acids released from adipose tissue into the bloodstream
second-messenger pathway. in response to growth hormone are converted in the liver to
ketone bodies for release into the circulation. Growth hor-
mone also stimulates fatty acid uptake in muscle, further
Growth hormone promoting the utilization of fatty acids as an energy source.
The production and release of growth hormone (GH)in the By increasing the utilization of fatty acids, growth hormone
anterior pituitary gland is under the direct control of GH- helps conserve muscle glycogen stores (see Figure 9-28).
HORMONES: REGULATION A N D A C T I O N 335
...............................................................................
Figure 9-28 Many of the actions of growth hormone are antagonistic to for ketone body synthesis.The GH-induced general depression of glu-
those of insulin. Output of insulin from pancreatic beta cells occurs in re- cose uptake (except in the central nervous system) leads to a rise in
sponse to high blood glucose, as after a meal. Growth hormone (GH) is plasma glucose, which then stimulates insulin secretion. The insulin
released, usually several hours after a meal or after prolonged exercise, stimulates glucose uptake into cells and thus counteracts GH-induced
in response to insulin-induced hypoglycemia. Growth hormone causes hyperglycemia.
lipolysis and fatty acid uptake by muscle tissue for energy and by the liver
In contrast to insulin, which causes a decrease in blood amino acids, and fatty acids) have begun to decrease.
glucose levels, growth hormone causes an elevation of Furthermore, growth hormone stimulates insulin secretion
blood glucose. Therefore, growth hormone counteracts both directly, through its action on the pancreatic beta
hypoglycemia, whereas insulin counteracts hypeglycemia. cells, and indirectly, through its effect in elevating plasma
Growth hormone elevates blood glucose by three mecha- glucose levels.
nisms: it stimulates gluconeogenesis from fat, blocks Growth hormone also stimulates RNA and protein
glucose uptake by tissues other than the nervous system, synthesis, which may account for its developmental effects
and promotes the utilization of fatty acids in place of glu- in promoting the growth of tissues-in particular, cartilage
cose. Thus both glucagon, which stimulates glycogen and subsequently bone. GH-stimulated tissue growth oc-
breakdown in the liver, and growth hormone act to main- curs by an increase in cell number (i.e., cell proliferation)
tain adequate blood glucose levels. Growth hormone rather than an increase in cell size. As noted above, thyroid
reaches its peak plasma level several hours after a meal, hormones and growth hormone work synergistically to
when immediate energy supplies (e.g., blood glucose, promote tissue growth during development. The growth-
enhancing effects of growth hormone depend very much on Mammals produce arginine vasopressin, but other ver-
the stage of development of the animal: the neonatal mam- tebrates produce slightly different nonapeptides with simi-
mal is relatively insensitive to growth hormone but be- lar actions, as noted earlier. Reptiles, fish, and birds pro-
comes more sensitive as it grows. GH not only stimulates duce a related peptide, called arginine vasotocin, which
proliferation of cells directly, it also stimulates the liver to exerts effects similar to those of vasopressin and oxytocin
produce other growth-promoting factors, called insulin-like (see Table 9-4). Like vasopressin, vasotocin promotes wa-
growth factors, that also act directly on cells to promote ter resorption by the animal. In addition, this hormone may
growth. Disturbances in the secretion of growth hormone play a role in sexual behavior and is associated with expul-
lead to several patterns of abnormal growth and develop- sion of eggs from the oviduct in turtles (somewhat analo-
ment in humans: gous to oxytocin action). Both vasopressin and vasotocin
are known to cause smooth muscle contraction. ADH and
Gigantism: excessive size and stature caused by hyper- its analogs exert their effects through the cAMP pathway.
secretion of growth hormone during childhood (before The mineralocorticoids, in particular aldosterone, en-
puberty hance resorption of sodium (and, indirectly, chloride) by
Acromegaly: enlargement of the bones of the head and the distal tubules and the collecting tubules of the kidney,
of the extremities caused by hypersecretion of growth thereby increasing the osmolarity of the blood. Aldosterone
hormone beginning after maturity is one of the steroid hormones secreted by the adrenal
cortex under the stimulation of ACTH (adrenal corti-
Dwarfism: abnormal underdevelopment of the body cotrophic hormone). Secretion of aldosterone is stimulated
caused by insufficient secretion of growth hormone by angiotensin I1 and high blood K+ and is subject to neg-
during childhood and adolescence ative feedback by the hormone on the CRH-secreting neu-
rons of the hypothalamus and on the ACTH-secreting cells
Little is known about the cell-surface receptors that of the anterior pituitary gland (see Figure 9-6). The miner-
bind growth hormone or the intracellular signaling path- alocorticoids, like other steroid hormones, mediate their
ways stimulated by hormone binding. However, applica- effects by binding to intracellular receptors and modifying
tion of growth hormone to tissues from young animals has gene expression.
been shown to inhibit adenylate cyclase activity and hence Atrial natriuretic peptide (ANP) acts on the kidney to
lead to a decrease in cAMP levels. reduce sodium and, therefore, water resorption, leading to
an increase in urine production and sodium excretion in the
Hormones That Regulate Water and kidney. Thus the effects of this hormone counteract those
Electrolyte Balance of aldosterone and ADH. ANP is released by the atrium of
The major organs involved in regulation of water and elec- the heart into the blood in response to an increase in venous
trolyte balance in vertebrates are the kidney, intestine, and pressure. Its mechanism of action is not clear.
bone, and in fish, the gills. Since epithelial cells are respon- As we saw earlier, Ca2+plays a key role as a second
sible for the uptake or excretion of water and electrolytes, messenger and regulatory agent in the cell. Thus careful
most of the hormones that regulate water and electrolyte regulation of the concentration of Ca2+in the blood and
balance act on these epithelial tissues. The processes for the extracellular fluid is critical. This ion is actively ab-
maintaining water and electrolyte balance are described sorbed through the intestinal wall into the plasma and is de-
in more detail in Chapter 14. Here we consider the hor- posited in bone, the major depot for storage of Ca2+.Elim-
mones that play a major role in regulating these processes ination of Ca2+from the body occurs through the kidney.
(Table 9-8). The balance of these processes, which determines the blood
Antidiuretic hormone (ADH),also called vasopressin, Ca2+ concentration, is influenced by three hormones:
regulates water turnover in the mammalian kidney. Secre- parathyroid hormone, calcitonin, and calcitriol.
tion of this neurohormone from the posterior pituitary Parathyroid hormone (PTH),also known as parathor-
gland is stimulated by high blood osmolarity, acting on 0s- mone, is secreted from the paired parathyroid glands in
moreceptors in the anterior hypothalamus (seeFigure 9-7). response to a drop in plasma Ca2+ levels. It acts to in-
By increasing the water permeability of the kidney collect- crease plasma Ca2+by promoting Ca2+mobilization from
ing duct, ADH stimulates resorption of water from the bone, increasing Ca2+uptake from the forming urine in
forming urine; the end result is a reduction in urine volume kidney tubules, increasing renal P043- excretion, and en-
and increased water retention by the body. Increases in ve- hancing intestinal Ca2+ absorption (Figure 9-29). PTH
nous blood pressure, reflecting increases in blood volume, works in conjunction with calcitriol, a steroid-like com-
stimulate atrial stretch receptors in the heart; these then pound produced from vitamin D ingested with some
send an inhibitory signal to the hypothalamus that de- foods and from vitamin D,, which can be synthesized
creases ADH release and, therefore, enhances urine pro- from cholesterol in the skin. Conversion of these precur-
duction, leading to a reduction in blood volume. ADH also sors into calcitriol involves reactions in the liver and
enhances the release of ACTH and TSH from the anterior kidneys. The actions of calcitriol are similar to those of
pituitary gland. parathyroid hormone.
HORMONES: REGULATION A N D A C T I O N 337
...............................................................................
TABLE 9-8
Mammalian hormones.involved in regulating water and electrolyte balance
Hormone Tissue of origin Structure Target tissue Primary action Regulation
in renal tubules
Mobilization of Absorption of Ca2+
Ca2+from bone Excretion of Pod3- from intestine
mediated by calcitriol
Figure 9-29 Calcitonin and parathyroid hormone (PTH) have opposite of Ca2+in the blood stimulate parafollicular cells in the thyroid gland to
effects on plasma Ca2+levels in mammals. Low levels of plasma Ca2+ release calcitonin, which acts to increase plasma Ca2+.Calc~triol, the ac-
stimulate the cells of the parathyroid glands to release PTH, which has tive hormonal form of vitamin D, also increases intestinal absorption of
several actions all tending to Increase plasma Ca2+.High concentrations Ca2+.
338 P H Y S I O L O G I C A L PROCESSES
. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
TABLE 9-9
Important mammalian reproductive hormones
Other Hormones
Oxytocin Neurohypophysis Nonapeptide Uterus, Promotes smooth muscle Cervical distention and suckling
mammary glands contraction and milk ejection stimulate release; high
progesterone inhibits release
Prolactin (PL) Adenohypophysis Peptide Mammary glands Increases synthesis of milk Continuous secretion of
(alveolar cells) proteins and growth of PL-inhibiting hormone (PIH)
mammary glands; elicits normally blocks release;
maternal behavior increased estrogen and
decreased PIH secretion permit
release
Calcitonin is secreted from the parafollicular, or C, cells dition to the steroid sex hormones, two peptide hormones
in the thyroid gland in response high plasma Ca2+levels. produced in the pituitary gland function in parturition and
It rapidly suppresses Ca2+loss from bone, qulckly coun- lactation. Table 9-9 summarizes the properties of the
tering the effects of PTH. Although calcitonin and PTH steroid and peptide reproductive hormones.
have opposing actions on bone metabolism, there is no The production and secretion of the steroid sex hor-
feedback interaction between them. Each hormone, how- mones in both males and females are promoted by folli-
ever, exerts negative feedback on its own secretion. The cle-stimulating hormone (FSH) and luteinizing hormone
dominance of calcitonin prevents hypercalcemia and ex- (LH), which are synthesized in the anterior pituitary
tensive dissolution of the skeleton. Essentially, then, bone (see Table 9-3). These tropic hormones are released from
acts as a large reservoir and buffer for Ca2+and also PO:-. the anterior pituitary gland in response to the hypothala-
The plasma Ca2+and PO:- levels are held within narrow mic gonadotropin-releasing hormone (GnRH). The
limits by the opposing actions of PTH and calcitonin, steroid sex hormones exert negative feedback on the
which regulate the flux of these minerals between plasma GnRH-secreting neurons of the hypothalamus and on
and bone. the anterior pituitary endocrine cells that produce FSH
PTH and calcitonin are both peptide hormones and and LH.
bind to cell-surface receptors. Little is known about the
intracellular signaling pathways mediating their effects. Steroid sex hormones in males
Calcitriol is lipid soluble and presumably binds to an intra- The seminiferous tubules of the mammalian testes are lined
cellular receptor. with germ cells and Sertoli cells (Figure 9-30). Binding of
FSH to receptors on Sertoli cells stimulates spermatogene-
Reproductive Hormones sis in the germ cells after sexual maturity, either continu-
In vertebrates several steroid hormones that affect repro- ously or seasonally, depending on the species. The cells of
duction (the estrogens, the androgens, and progesterone) Sertoli support development of the sperm and are respon-
are produced in the gonads (testis or ovary) and adrenal sible for synthesis of androgen-binding protein (ABP)and
cortex of both sexes from cholesterol (see Figure 9-23). inhibin. Lying between the seminiferous tubules are inter-
Cholesterol is first converted to progesterone, which is then stitial cells, called Leydig cells, which produce and secrete
transformed into the androgens (androstenedione and sex hormones, particularly testosterone. Both testosterone
testosterone). These are then converted into the estrogens, itself and inhibin provide inhibitory feedback to the hypo-
of which estradiol-17P is the most potent. The steroid sex thalamic centers controlling GnRH production and hence
hormones, like other steroid hormones, bind to intracellu- diminish release of the gonadotropins FSH and LH from
lar receptors and modify expression of specific genes. In ad- the anterior pituitary gland.
HORMONES: REGULATION AND ACTION 339
...............................................................................
Low blood testosterone; neural input
feedback
Semln~feroustubule
(spermatogenes~s)
Cells of Sertoll
Figure 9-30 Testosterone, the primary sex hormone in males, has nu- follicle-stimulating hormone (FSH) and luteinizing hormone (LH). Some
merous actions and is regulated by neural stimuli and feedback control. of the actions of testosterone are ind~catedat the bottom of the figure.
A decrease in blood testosterone stimulates the secretion of go- High testosterone and inhibin, which also is secreted by the testes, in-
nadotropin-releasinghormone (GnRH),which promotes the release of hibits FSH secretion both directly and indirectly.
The estrogens and androgens are important in both Steroid sex hormones in females: regulation of the
sexes in various aspects of growth, development, and mor- menstrual cycle
phologic differentiation, as well as in the development and Unlike androgens, which stimulate prenatal differentiation
regulation of sexual and reproductive behaviors and cycles. of the embryonic male genital tract, the estrogens play no
However, androgens predominate in the male, whereas es- such role in early differentiation of the female tract. How-
trogens predominate in the female. The androgens trigger ever, estrogens stimulate later development of primary sex-
development of the primary male sexual characteristics (e.g., ual characteristics such as the uterus, ovary, and vagina.
the penis, vas deferens, seminal vesicles, prostate gland, epi- The estrogens also are responsible for development of the
didymis) in the embryo and the secondary male sexual char- secondary female sex characteristics such as the breast and
acteristics (e.g., the lion's mane, the rooster's comb and for regulation of reproductive cycles (Figure 9-31).
plumage, and facial hair in men) at the time of puberty. The Simultaneous reproduction within an entire popula-
androgens also contribute to general growth and protein tion can be of obvious survival value to a species. The
synthesis-in particular, the synthesis of myofibrillar pro- gathering of large numbers of individuals of both sexes for
teins in muscle, as evidenced by the greater muscularity of mating, bearing young, and parenting the young during
the males relative to the females in many vertebrate species. this period of high vulnerability can be timed to coincide
Neural input
High
levels
only
Figure 9-31 Estrogens and progesterone, the primary steroid sex hor- ulate release of luteinizing hormone (LH), which triggers ovulation and
mones in females, mediate reprodudive cycles and other effects under subsequent development of the corpus luteum. The corpus luteum se-
complex regulation. In mammals a decrease in progesterone and estro- cretes primarily progesterone and estrogens, which are needed to maln-
gen levels, as well as neural inputs, stimulates release of gonadotropin- tain pregnancy. Eventually, the high levels of FSH and LH, as well as pro-
releasing hormone (GnRH).This acts on the anterior pituitary gland, stim- gesterone, inhibit the activity of the hypothalamic neurosecretory cells,
ulating secretion of follicle-stimulating hormone (FSH), which promotes leading to a decrease in gonadotropin secretion. This prevents menstrual
development of the primordial follicles in the ovary. Estrogens secreted cycling during pregnancy.
by the follicles and by the interstitial cells eventually reach levels that stim-
Ovulation Ovulation
4
Follicular phase
--
F
Luteal phase
0 14 28 14 28
(0)
Time (days)
Figure 9-32 The primate menstrual cycle is regulated by periodic peak and then fall, initiating menstruation. The subsequent decrease in
changes in the levels of the gonadotropins, estrogens, and progesterone. estrogen, progesterone, and inhibin levels allows pituitary secretion of
Before ovulation, follicle-stimulating hormone (FSH) promotes matura- FSH and LH to increase again, thus initiating a new cycle. If implantation
tion of ovarian follicles, which secrete estrogen. High estrogen levels and pregnancy occur (right), secretion of chorionic gonadotropin (CG) by
cause a surge of luteinizing hormone (LH),which triggers ovulation from the placenta "rescues" the corpus luteum, which maintains secretion of
one follicle. LH promotes development of the corpus luteum, and in- estrogen and progesteronefor the first two to three months of pregnancy
duces the corpus luteum to secrete progesterone and some estrogen. In in the human. Thereafter, the placenta itself secretes estrogens and pro-
the absence of implantation (left),the progesterone and estrogen levels gesterone. [Adapted from McNaught and Callander, 1975.1
During the luteal phase, which begins with ovulation, cle, but in the nonprimate mammals the luteal phase is
estrogen secretion declines and LH transforms the ruptured much shorter. The number of cycles per year also varies
follicle into a temporary endocrine tissue, the corpus luteum. among species. The human menstrual cycle of approxi-
The corpus luteum secretes estrogen and progesterone, mately 28 days normally occurs 13 times a year. Among
which exert negative feedback on GnRH release by the hy- nonprimate mammals, some have only one cycle per year
pothalamus, leading to decreased secretion of FSH and LH. (usually in spring); others, such as the laboratory rat, have
The ovarian hormone inhibin, which is released along with multiple cycles throughout the year.
the ovum, acts on the anterior pituitary, inhibiting FSH (but During gestation progesterone and estrogen, secreted
not LH) release. Progesterone stimulates secretion of en- from the corpus luteum or placenta, initiate growth of the
dometrial fluid by the endometrial tissue, preparing it for mammary tissues in preparation for lactation. Prolactin
implantation of a fertilized ovum. In the absence of fertil- and placental lactogen, a hormone produced in the pla-
ization and implantation of an ovum, the corpus luteum de- centa, also aid in preparing the mammary glands for lac-
generates after a 14 ? 1-day period (in humans), and secre- tation, but synthesis of milk is inhibited by progesterone
tion of estrogen and progesterone subsides. In humans and during pregnancy. The negative feedback of estrogen and
some other primates, this precipitates the menses, or shed- progesterone on the hypothalamus and anterior pituitary
ding of the uterine lining. With the reduction in estrogen, gland prevent release of FSH and LH during pregnancy,
progesterone, and inhibin levels, FSH and LH secretion by thereby preventing ovulation. Birth control pills contain
the pituitary increases again, initiating a new cycle. small amounts of progesterone and estradiol or their syn-
If the released ovum is fertilized as it travels down the thetic analogs. Taken daily, these steroids mimic the earli-
ciliated fallopian tube and the fertilized ovum becomes est stages of pregnancy, preventing ovulation and also act-
implanted in the endometrium of the placental mammal, ing on the endometrium, thereby providing a highly
the developing placenta begins to produce chorionic effective means of avoiding conception.
gonadotropin (CG) (see Figure 9-32, right). This hormone,
whose action is similar to LH, induces further growth of Hormones involved in parturition and lactation
the active corpus luteum, so that estrogen and progesterone As pregnancy nears term, cervical distention stimulates re-
secretion continues. The placenta begins secreting CG lease of oxytocin from the posterior pituitary gland (see
within about a day of implantation of the ovum and effec- Table 9-9). This hormone induces contractions of the
tively takes over the gonadotropic function of the pituitary smooth muscle in the uterine wall, which are critical to the
during early pregnancy by maintaining the corpus luteum. normal birth process (parturition). Certain prostaglandins
Pituitary FSH and LH are not secreted again until after par- also may stimulate uterine contractions during childbirth.
turition (birth of the fetus). In many mammals, including After parturition, a decrease in progesterone levels relieves
humans, the corpus luteum continues to grow and to se- inhibition of the milk-synthesizing machinery, permitting
crete estrogen and progesterone until the placenta fully lactation to begin. Milk production is mediated by pro-
takes over the production of these hormones, at which time lactin, along with the glucocorticoids, and the release of
the corpus luteum degenerates. In other mammals, such as milk is induced by oxytocin. Both prolactin and oxytocin
the rat, continued secretion by the corpus luteum, stimu- are released during suckling as a result of neural input to
lated by prolactin, is essential to the maintenance of preg- the hypothalamus arising from stimulation of the nipples.
nancy throughout its term.
The durations of the follicular and luteal phases of the Prostaglandins
reproductive cycle vary among different mammalian The long-chain, unsaturated, hydroxy fatty acids called
groups. They are about equal in the primate menstrual cy- prostaglandins were first discovered in the 1930s in semi-
TABLE 9-10
Selected prostaglandins
n-
Pupa 1
Cement
Glass
tube
Pupa 2
Figure 9-33 Parabiosis, the joining of body parts from different individ-
uals, is a useful experimental method in insect endocrinology. Insect tis-
sues readily survive such radical surgery as transection and decapitation.
EFImm -+P-Ecdysone
Target cells
In this example, the abdomen of one pupa is joined to another pupa
through a glass tube. Glass windows at either end permit visual inspec- Figure 9-34 Of the five major insect developmental hormones, three are
tion of the developing tissues. produced by neurosecretory cells and two by endocrine tissues. Neu-
rosecretory cells in the brain synthesize prothoracicotropic hormone
(PTTH) and eclosion hormone, which are stored in nerve terminals until
their release into blood sinus spaces in the corpus cardiaca and corpus
Ecdysone, produced by the prothoracic glands, is syn- allata, two paired neurohemal organs. A third neurohormone, bursicon,
thesized from cholesterol. It is structurally similar to is released primarily from nerve terminals in the nerve cord. The corpus
allatum also contains nonneural cells that elaborate juvenile hormone
vertebrate steroid hormones but contains more hy-
(JH). Under the stimulus of PTTH, the prothoracic gland produces and se-
droxyl groups (Figure 9-35B). cretes a-ecdysone, which is converted to the active molting hormone P-
Eclosion hormone, a peptide neurohormone, is released ecdysone. [Adapted from Riddiford and Truman, 1978.1
from neurosecretory cells whose terminals are in the
corpora cardiaca, which are paired neurohemal organs
immediately posterior to the brain. factor, a-ecdysone. Insects require cholesterol in their diets
Bursicon, also a neurohormone, is produced by other to synthesize this steroid hormone. It is now thought that
neurosecretory cells in the brain and nerve cord. It is a a-ecdysone is a prohormone converted to the physiologi-
protein with a molecular weight of about 40,000. cally active form, 20-hydroxyecdysone (P-ecdysone), in
several peripheral target tissues (see Figure 9-35B).
PTTH is shipped by axoplasmic transport along the ax- Juvenile hormone, acting in association with p-
ons of the neurosecretory cells to storage depots, or neuro- ecdysone, promotes the retention of the immature ("juve-
hemal organs, formed by the terminals of the axons (see nile") characteristics of the larva, thereby postponing meta-
Figure 9-34). The corpus cardiacum was thought to be the morphosis until larval development is completed. The
neurohemal organ that stores and releases PTTH, but more presence of juvenile hormone in the early nymphal instar
recent evidence from the tobacco hornworm moth, Man- was demonstrated in the mid-1930s in experiments by V. B.
duca sexta, indicates that the axons of the PTTH-produc- Wigglesworth in which parabiotic coupling of the early in-
ing neurosecretory brain cells actually pass through the car- star to a final instar prevented the latter from becoming an
pus cardiacum and end within the corpus allatum, which is adult. The circulating concentration of juvenile hormone is
located at the posterior end of the corpus cardiacum. Thus, highest early in larval life, dropping to a minimum at the
the corpus allatum appears to be the site at which the neu- end of the pupal period (Figure 9-36). Metamorphosis to
rosecretory endings release PTTH into the blood. It re- the adult stage occurs when juvenile hormone disappears
mains to be determined if this is true of all insects. from the circulation. The concentration then rises again in
After its release into the blood, PTTH activates the pro- the reproductively active adult. In the males of some insect
thoracic gland to synthesize and secrete the molt-inducing species, juvede hormone promotes development of the
HORMONES: REGULATION A N D ACTION 345
TABLE 9-1 1
Insect developmental hormones
Juvenile hormone o
c
.-
+-
m
Stage of development
Figure 9-35 Juvenile hormone and pecdysone play key roles in regu- Figure 9-36 Normal progression through the insect life cycle depends
lating insect development. (A) The structure of juvenile hormone from on changes in the level of juvenile hormone. Metamorphosis of the juve-
the cecropia moth Hyalophora cecropia. This hormone promotes the re- nile larval form to the pupa occurs when the concentrationof juvenile hor-
tention of iuvenile characteristicsin larvae and induces reproductive mat- mone falls below a certain threshold level. After the adult insect emerges
uration in adults. Several hornologs of juvenile hormone occur naturally and feeds, secretion of juvenile hormone beglns agaln, regulating ovar-
in insects. (B) The structure of P-ecdysone, the physiologically active ian activity and stimulating development of male accessory organs.
molt-inducing hormone. The prohormone a-ecdysone, which lacks the [Adapted from Spratt, 1971.]
hydroxyl group on C-20 (red), is synthesized from cholesterol in the pro-
thoracic glands of insects. After its release, a-ecdysone is converted in
certain target tissues into the active hormone pecdysone.
accessory sexual organs; in many female insects, it induces Epidermal
yolk synthesis and promotes maturation of the eggs. cells
Thus, the normal development of an insect depends on Ecdysone \ Cuticle
precisely adjusted concentrations of juvenile hormone at
each stage. The role of this hormone is somewhat analo-
gous to that of thyroid hormones in the regulation of
amphibian development. In both cases, disturbance of the High JH Bursicon
relationship between hormone concentration and devel-
opmental stage leads to abnormal development. Because of
its potency in preventing maturation in insects, juvenile
Ecdysone
-
--- -- - Thickened cuticle
\ \pupa
hormone and its synthetic analogs are promising as poten-
tial nontoxic, ecologically sound means of combating insect
pests, and one against which the insect would find it diffi- Low JH Bursicon
cult to develop resistance.
During growth and development of insects, the epider-
-
Ecdysone Bristles
mis undergoes conspicuous changes, including production
of the cuticle, the chitinous, horny outer covering. There-
fore, considerable attention has been given to the produc-
tion of new cuticle, its tanning, and the shedding of the old
m-m+.B-m-m
No JH
ICU
Eclosion Bursicon
cuticle during molting. PTTH, juvenile hormone, and P- hormone
ecdysone are all involved in the initiation of molting (Figure Apolysis Cuticle Ecdysis Tanning
9-37). Ecdysone, secreted by the prothoracic glands in re- formation
sponse to stimulation by PTTH, acts on the epidermis to Figure 9-37 The cuticularchangesinvolved in molts leading to larval, pu-
initiate production of the new cuticle, which begins with pal, and adult stages are controlled by the level of juvenile hormone (JH).
apolysis, the detachment of the old cuticle from the under- Ecdysone initiates productionof new cuticle, beginningwith detachment
of the old cuticle (called apolysis). Shedding of the old cuticle (called ec-
lying epidermal cells. The epithelial cells then begin synthe-
dysis) is triggered by eclosion hormone. Although bursicon regulates
sizing the materials for the new cuticle, while the old cuticle hardening and darkening (tanning)of the new cuticle, the JH concentra-
is partially digested from beneath by enzymes in the molt- tion determines whether it has larval, pupal, or adult characteristics (top
ing fluid secreted by the epidermis. At high concentrations to bottom).[Adaptedfrom Riddiford and Truman, 1978.1
of juvenile hormone a larval-type new cuticle is formed,
whereas at low concentrations levels an adult-type cuticle is
produced and other events of metamorphosis ensue.
Two additional hormones, eclosion hormone and bur-
SUMMARY
sicon, are responsible for promoting the terminal phase of The physiological and biochemical processes occurring in
the molting process. Shedding of the cuticle of the pupae, cells, tissues, and organs are controlled and coordinated in
termed ecdysis, is triggered by eclosion hormone at least in animals in large part by special blood-borne messenger
some holometabolous species. The pale, soft cuticle of a molecules termed hormones, which are released from en-
newly molted insect is expanded by respiratory movements docrine secretory tissues. In vertebrates these hormones fall
of the insect to the next size before it hardens, or tans, un- into four chemical categories: (1) amines, (2) prosta-
der the influence of bursicon (see Figure 9-37). glandins, (3)steroids, and (4) peptides and proteins. After
Figure 9-38, on the opposite page, outlines the hor- a hormone is released from its site of origin, it circulates at
monal interactions that regulate metamorphosis of low concentration in the bloodstream throughout the body.
Hyalophora cecropia, a holometabolous insect. PTTH re- The selective actions of hormones on specific target tissues
lease initiates larval ecdyses (molts)and stimulates the pro- depends on the preferential distribution among tissues of
thoracic gland to secrete the molting hormone ecdysone. hormone-specific receptors and various effector proteins
Growth continues through a series of instars, which remain that mediate hormone-induced cellular responses.
larval as long as the concentration of juvenile hormone re- The secretion of hormones from endocrine tissues is
mains above a minimum. This process of growth and molt- stimulated either by hormones released from other edocrine
ing is usually completed in four or five instars, during which tissues or by neurohormones released from specialized neu-
the concentration of juvenile hormone progressively de- rons; the latter form the basis for neuroendocrine reflexes.
clines. Once the juvenile-inducing effects of juvenile hor- In addition, some endocrine tissues respond directly to con-
mone are removed, the larva molts to the pupal stage. The ditions of the extracellular environment. The secretory ac-
pupa is the over-wintering stage of the cecropia moth, caus- tivities of most endocrine tissues are modulated by negative
ing it to undergo obligatory diapause. Prolonged exposure feedback; that is, the increasing concentration of the hor-
to cold stimulates release of PTTH in the pupa, inducing mone itself, or a response to the hornione by the target tis-
the release of ecdysone; in the absence of juvenile hormone, sue (e.g., reduced blood glucose levels in the insulin loop),
ecdysone induces pupal development into the adult moth. has an inhibitory effect on the synthesis or release of the
HORMONES: REGULATION AND ACTION 347
...............................................................................
BRAIN
(neurosecretory cells)
\
cardiacum
Corpus
allaturn
Low concentration JH
a PTTH stimulates
High concentration JH
causes larval molting
Ecdysone (ec)
LARVA
PUPA (d~apause) EGGS @$
Low temperature
stimulates P l T H
Figure 9-38 Interactions of juvenile hormone and ecdysone regulate velopmental sequence of the cecropia moth. See text for discussion.
metamorphosis in holometabolousinsects. This example shows the de- [Adapted from Spratt, 1971.]
same hormone. Positive feedback occurs in some systems; to convert ATP into the second messenger CAMP.Binding
occasionally hormone secretion is feedforward (i.e., not of some hormones stimulates guanylate cyclase to produce
regulated by any consequences of the secreted hormone). the related second messenger cGMP, in most cases by a
To exert their effect, all hormones must bind to specific somewhat different sequence of steps. Once formed, cyclic
receptors; this binding initiates intracellular mechanisms nucleotide activates a specific protein kinase, which then
leading to a cellular response(s).Steroid hormones and thy- phosphory-lates various effector proteins mediating cellu-
roid hormones, being lipid soluble, enter cells freely and lar responses.
bind to receptor proteins present in the cytosol. The result- In the inositol phospholipid signaling system, hormone
ing hormone-receptor complexes translocate into the nu- binding to a G protein-linked receptor activates phospho-
cleus, where they bind to regulatory elements in the DNA inositide-specific phospholipase C, which then hydrolyzes
thereby stimulating (and in a few cases inhibiting) tran- PIP, into two major second messengers, InsP3 and DAG.
scription of specific genes. All other hormones bind to cell- InsP, induces release of Ca2+from intracellular stores. In
receptors located in the plasma membrane of target cells. addition, InsP, is converted to InsP4, which promotes entry
This binding triggers one or more intracellular signal-trans- of Ca2+from the cell exterior into the cell. The resulting in-
duction pathways leading to the cell's responses. crease in free cytosolic Ca2+regulates the activity of a va-
In the CAMPsignaling system, hormone binding acti- riety of cellular proteins. DAG, on the other hand, remains
vates a G protein, which then stimulates adenylate cyclase in the membrane and activates membrane-bound protein
kinase C. This kinase then phosphorylates various effector REVIEW QUESTIONS
proteins, which are responsible for the cellular responses.
In the Ca2+signaling system, hormone stimulation of 1. Give three examples of chemical regulation that
the receptor directly activates Ca2+channels in the plasma do not involve the secretion of specific hormone
membrane, thereby stimulating Ca2+ influx. Hormone- molecules.
induced changes in intracellular Ca2+levels regulate diverse 2. What criteria must be met before a tissue can be
cellular processes. In membrane-enzyme signaling systems, unequivocally identified as having an endocrine
hormone binding activates intrinsic enzyme activity in the function?
cytosolic domain of the receptor. The activated enzymes, in 3. Give examples of short-loop and long-loop negative
turn, induce intracellular responses. feedback in the control of hormone secretion.
Even in the same cell, a single hormone may bind to dif-
4. Discuss two examples that illustrate the intimate
ferent cell-surface receptors linked to different second
functional association of the nervous and endocrine
messengers, thereby inducing the same cell response (con-
systems.
vergent pathways) or different responses (divergent path-
ways). A particular receptor may be coupled to two differ- 5. Explain how it is possible that the actions of epi-
ent G proteins, each linked to its own second-messenger nephrine and glucagon are similar but their actions
pathway or both linked to the same second-messenger are confined to different tissues.
pathway. Other variations in signaling systems, some in- 6. How can a single second messenger (e.g., cAMP or
volving third messengers, also are possible. Clearly, intra- InsP3), induced by binding of different hormones,
cellular signaling pathways typically interact with each mediate different cellular responses in different
other in various ways to control hormone-induced intra- tissues?
cellular responses. 7. Explain how a small number of hormone molecules
Although most hormones have multiple actions, they can elicit cell responses involving millions of times as
can usefully be grouped into several functional classes plus many molecules.
the very diverse prostaglandins. The production and secre-
tion of several direct-acting hormones are regulated indi- 8. What is the significance of protein phosphorylation
rectly by hypothalamic releasing and inhibiting hormones in intracellular signaling systems?
and directly by tropic hormones produced in the anterior 9. How can a working muscle mobilize glycogen
pituitary gland. stores in the absence of epinephrine-induced glyco-
The following hormones have major roles in regulating genolysis?
metabolism and developmental processes: glucocorticoids 10. Describe two ways in which the concentration of free
and catecholamines, which are produced in the adrenal cytosolic Ca2+becomes elevated. Discuss the role of
glands and affect energy metabolism; thyroid hormones, Ca2+as a second and third messenger.
which regulate metabolic rate; insulin and glucagon, which
are produced in the pancreas and have opposite effects 11. Describe the similarities and differences that charac-
on blood glucose levels; and growth hormone, which is terize four intracellular signaling systems described in
produced in the anterior pituitary and works synergisti- this chapter.
cally with the thyroid hormones to promote growth and 12. Describe the interactions between CAMPand the in-
development. ositol phospholipid pathway.
The reproductive hormones include the androgens (in 13. Describe the interrelations between Ca2+and cAMP
males) and estrogens (in females),which promote develop- in the mammalian salivary gland and liver to illus-
ment of sexual characteristics and gametes (sperm or trate convergent and divergent second-messenger
oocytes). In females, progesterone acts to prepare the en- pathways activated in response to a single first mes-
dometrium for implantation and helps prepare breast tis- senger such as epinephrine.
sue for lactation; oxytocin stimulates uterine contractions 14. How do the glucocorticoids (also growth hormone
during birth and milk ejection after birth; and prolactin and glucagon) combat hypoglycemia?
promotes formation of milk and maternal behavior.
The hormones most responsible for regulating water and 15. What role do the thyroid hormones play in amphib-
electrolyte balance are antidiuretic hormone (ADH),which ian development?
increases water resorption in the kidney; mineralocorticoids, 16. How does insulin produce its hypoglycemic effects?
promotes Na+ resorption in the kidney; atrial natriuretic 17. What factors influence secretion of growth hormone?
peptide (ANP),which reduces Na+ and water resorption in
18. Discuss the endocrine control of the menstrual cycle.
the kidney; parathyroid hormone and calcitriol (derived
from vitamin D or cholesterol), which act to increase the 19. Explain how birth control pills prevent conception.
plasma Ca2+concentration; and calcitonin, which has the 20. Discuss the role of juvenile horinone in the develop-
opposite action, decreasing the blood Ca2+concentration. ment and metamorphosis of an insect.
HORMONES: REGULATION A N D ACTION 349
....................................... ......................................
SUGGESTED READINGS Robb, S., T. R. Cheek et al. 1994. Agonist-specific coupling
of a cloned Drosophila octopamineltyramine receptor
Alberts, B., D. Bray et al. 1994. Molecular Biology of to multiple second messenger systems. EMBO J.
the Cell. 3d ed. New York & London: Garland 13:1325-1330.
Publishing. Truman, J. W. 1992. The eclosion hormone system of in-
Hadley, M. E. 1992. Endocrinology. 3d ed. Englewood sects. Prog. Brain. Res. 92:361-374.
Cliffs, N.J.: Prentice Hall.
Raymond, J. R. 1995. Multiple mechanisms of receptor-G
protein signaling specificity.Am. J. Physiol. 269 (Renal
Fluid Electrolyte Physiol. 38):F141-F158.
A nimal movements-such as locomotion, eating, and
copulation-are generated by three fundamentally
different mechanisms: amoeboid movement, ciliary and fla-
Recent research on comparative and integrative aspects
of muscle function has uncovered unexpected diversity
among skeletal muscles and revealed an elegant matching
gellar bending, and muscle contraction. In addition, sound between the design of a muscle and its biological function.
production and nearly all other forms of communication Several examples of these varied deslgns are discussed in
that d o not rely on specific chemical signals are based on thls chapter.
muscle contractions. Most muscles contract when neurons
send signals to them, initiating a series of events that cause
STRUCTURAL BASIS OF MUSCLE
the muscles to generate force and become shorter. Muscle
CONTRACTION
contractions are the most apparent and dramatic macro-
scopic signs of animal life, and they have excited the imag- The general organization of skeletal muscle tissue is de-
ination of many people since ancient times. In the second picted in Figure 10-1. Muscles can move parts of an animal
century A.D., Galen hypothesized that "animal spirits" flow because each end of the muscle is attached to a bone or to
from nerves into muscles, inflating the muscles and in- some other structure, and when the muscle shortens, the
creasing their diameter at the expense of their length, caus- physical relationship between the anchor points changes.
ing them to shorten. Typically a muscle is anchored at each end by a tough strap
Even as recently as the 1950s, it was suggested that of connective tissue called a tendon. Each muscle consists
/
muscles shorten because linear molecules of "contractile of long, cylindrical, multinucleate cells (or muscle fibers),
proteins" within the muscles are caused to shorten. The hy- which are arranged in parallel with one another. This
pothesis stated that these molecules are helical in shape and arrangement allows all of the fibers in a muscle to pull in
that changes in the pitch of the helix produce changes in parallel with one another. Striated muscle fibers range from
length. This hypothesis was short-lived, however, because 5 to 100 p m in diameter, and may be many centimeters in
the development of new techniques in the 1950s led to dra- length. (Consider the length of the calf muscle fibers of a
matic advances in our understanding of muscle function. professional basketball player.) One reason for this extra-
Through evidence from electron microscopy, biochemistry, ordinary size is probably that each fiber arises from many
and biophysics, we have learned how the contractile mech- single embryonic muscle cells, called myoblasts, which fuse
anism of muscle is organized and how it produces force and during embryonic development to form multinucleated
shortening. It is also becoming clear how the process of units called myotubes. Each myotube contains many nuclei
contraction is initiated by electrical activity in the mem- within a single plasma membrane and differentiates into an
brane of muscle fibers. adult muscle fiber, sometimes called a myofiber. Each mus-
Muscles are classified, on both morphological and cle fiber is, in turn, composed of numerous parallel subunits
functional grounds, into two major types, smooth muscle called myofibrils, which consist of longitudinally repeated
I and striated muscle. One muscle type-vertebrate striated units called sarcomeres. The sarcomere is the functional
k. : muscle (primarily frog and rabbit skeletal muscle)-is the unit of striated muscle. The myofibrils of a muscle fiber are
best understood, and in this chapter we will consider it in lined up with the sarcomeres in register, so the fiber looks
i*
detail. Striated muscle itself can be subdivided into skele- banded, or striated, when it is observed with a light micro-
tal muscle and cardiac muscle. However, the mechanism by scope. This banded appearance gave rise to the name stri-
t
t
which all muscles contract is nearly identical, and the ma- ated muscle.
jor differences between the classes are found in their cellu- The structure of striated muscle fibers provides an ele-
lar organization. gant example of structure as the basis of function. The
352 PHYSIOLOGICAL PROCESSES
...............................................................................
Muscle Figure 10-1 All vertebrate skeletal muscles are
organized in a stereotyped hierarchy. The organ
called a muscle consists of parallel multinucleate
fibers, each of which contains many myofibrils.
Muscles are attached to bones or other anchor
points through tough connective tissue bands
called tendons. Each muscle fiber is derived em-
bryonicallyfrom a group of myoblasts that fuse to
form myotubes. A myotube then synthesizes the
proteins characteristic of muscle fibers and differ-
entiates into its adult form. The myofibrils are
made up of sarcomeres, arranged end-to-end.
Each sarcomere contains thin filaments of actin
and thick filaments of myosin, which Interdigitate
in a precise geometric relationship (see Figure 10-
3). The thin filaments are anchored in regions
called Z disks. [Adapted from Lodish et al., 1995.1
One myofibril
Z disk
I/ Sarcomere ,,
I M Ane I 1 band
I ~dtin ~ybsin I
disk thin filament thick filament Z disk
r
electron micrograph in Figure 10-2 shows a longitudinal ergy metabolism (e.g., creatine kinase). The portion of the
Y
section of several myofibrils. Each sarcomere is bounded at sarcomere between two A bands is called the I band (so
either end by a Z line (or Z disk) which contains a-actinin, named because this region is isotropic and does not polar-
one of the proteins found in all motile cells. Extending in ize light).
both directions from the Z line of a myofibril are numerous If cross-sections are made through the various regions
thin filaments consisting largely of the protein actin. These of a single sarcomere, the precisely arranged geometric re-
thin filaments interdigitate with thick filaments made up of lationship between thick and thin filaments is revealed
the protein myosin. Interdigitated thick and thin filaments (Figure 10-3).Only actin-containingthin filaments are seen
make up the densest portion of the sarcomere, the A band when the section is made through an I band, and only
(SO named because this band is anisotropic and strongly po- myosin-containing thick filaments are seen in a section
larizes visible light). The lighter portion in the center of the through an H zone. In the region of overlap, each myosin
A band is called the H zone, which contains only thick fil- filament is surrounded by six thin filaments, and it shares
aments. In the middle of the H zone is the M line, which has these actin filaments with surrounding thick filaments. Each
been shown to contain enzymes that are important in en- actin filament is surrounded by three myosin filaments.
I
MUSCLES A N D A N I M A L MOVEMENT 353
..............................................................................
,Glycogen Figure 10-2 The sarcomeres within different myofibrils are
aligned in register producing the characteristic bands of skele-
tal muscle. Electron micrographs not only reveal these bands
but also the sarcomere components giv~ngrise to them. This
electron micrograph of a longitudinal section through a frog
skeletal muscle includes two complete sarcomeres of three my-
ofibrils. The various bands and the Z disks (which appear as
lines in longitudinal section) are labeled. Dark granules be-
tween fibrils are glycogen. [Courtesyof L. D. Peachey.]
When a section through a sarcomere is examined at filament occur in groups of three; in each group, the cross-
high magnification with an electron microscope, small bridges are spaced about 14.3 nm apart and the angular dis-
projections, called cross-bridges, are visible; these pro- placement around the filament between successive cross-
jections extend outward from the myosin filaments and bridges is 120" (Figure 10-4B).This arrangement results in
make contact with the actin filaments during contraction each thick filament having nine rows of cross-bridges, with
(Figure 10-4A).The cross-bridges along the axis of the thick adjacent cross-bridges in a row sepapted by 43 nm.
B
Actin Troponin Tropomyosin
I complex
I I
I I
40 nrn
I
s
8
Thick be formed between actin and myosin. Tension
I '
myofilarnent , drops off with increased lenqth,
*
I rnyofilament
I - because the thick
and thin filaments overlap less and fewer cross-
bridges can be formed. It also drops off with de-
creased length, because thin filaments begin to col-
lide with one another, preventing further shortening.
Skeletal muscle rarely operates over such a broad
range of lengths, because the structure of the bones
and joints limits the range of movement, so sarcom-
ere length normally never departs significantlyfrom
the plateau region of this curve. [Adapted from Gor-
don et al., 1966.1
lengths. As the length of the sarcomeres is changed, so is the In fact, it was possible to predict some of the properties
amount of overlap between the actin and myosin filaments of the length-tensioncurve before the experiments had been
(Figure 10-8B).The sarcomere length was adjusted with the carried out. As noted above, the sliding-filament hypothe-
aid of an electromechanical system that controlled the ini- sis assumes that the force generated by a sarcomere is pro-
tial tension, allowing the sarcomeres to be held at any de- portional to the number of cross-bridges binding myosin
b sired constant length. Then the tension generated when the filaments to actin filaments. It also assumes that cross-
fiber was stimulated to contract was measured and plotted bridges are evenly distributed along each thick filament, ex-
as a function of sarcomere length. When the fiber was cept in the bare zone where no cross-bridges are present.
stretched until there was no overlap at all between thick (This second assumption has been experimentally verified.)
and thin filaments, stimulation produced no tension be- From these assumptions and the dimensions of the fila-
yond the passive elastic tension required to stretch the fiber ments given in Figure 10-9A, it is possible to predict the
to that length. When the fiber was held so that the actin fil- shape of the sarcomere length-tension curve.
aments overlapped completely with the segment of the
myosin filament bearing the cross-bridges, the tension gen- At what sarcomere length will the filaments be pulled
erated was maximal. When the fiber was so short that the beyond overlap and hence generate no force? To de-
actin filaments in the two halves of the sarcomere collided, termine the sarcomere length for a given amount of
the fiber-generated tension decreased with further shorten- overlap between filaments of known lengths, imagine
ing. Tension decreased still further if the fiber was so short a very tiny ant trying to crawl along the filaments
that the myosin filaments crumpled up against the Z Isks. from the center of one Z disk to the center of the next
358 PHYSIOLOGICAL PROCESSES
...........................................
I
I
I
! Thick filament (a)
I
I
, a = 1.6 urn
b= irn
actin-binding sites on the thin filaments (Figure 10-9B,
condition 2). As the sarcomere shortens farther, no
more cross-bridges are added to the number that can in-
teract with the thin filaments, so the force generated re-
mains the same. The end of the plateau occurs when the
thin filaments meet in the center of the sarcomere
(Figure 10-9B, condition 3).
Why does the force fall as the sarcomere continues to
shorten? The sliding-filamenthypothesis does not make
quantitative predictions about muscle force below the
point of maximal overlap, so it has been necessary to
answer this question experimentally. From one per-
spective, the force might be expected to remain con-
stant because all the cross-bridges overlap with actin
sites on the thin filaments and can, at least in theory,
generate force. However, two effects could reduce the
force generated. First, when the thin filaments overlap
at the middle of the sarcomere, binding of myosin
cross-bridges with the thin filaments could be sterically
hindered (Figure 10-9B, condition 4). Second, cross-
bridges might bind with an inappropriate thin filament
(one projecting from the Z disk at the other side of the
sarcomere) and exert a force that pushes the Z disks
% z + a + YZZ= 1.65 yrn apart, rather than pulls them together. Such a force
+ * would be considered negative and would need to be
subtracted from the force generated by the normal
cross-bridges.
filaments are known, the sar-
Figure 10-9 If the lengths of thlck and t h ~ n Why does force decline steeply a t 1.65 pm and fall
comere length can be predicted for varlous amounts of overlap (A) FII- to zero a t about 1.25 pm? The force generated de-
ament lengths measured from h~gh-resolut~on electron micrographs of
clines steeply when the sarcomere is so short that the
frog musclef~bersa, length of thlckf~lament,b, length of thln filament,
c, length of bare zone that IS free of cross-br~dgesat center of thtck flla-
thick filaments contact the Z disks at both ends of the
ment, z, thickness of Z dlsk (B) Amounts of overlap between thlck and sarcomere (Figure 10-9B, condition 5). At this point,
thln f~bersat lnflectlons In the sarcomere length-tens~on curve Each con- any further shortening of the sarcomere would require
d~tlonIn thlsf~gureIS matched to a polnt on the curve In Flgure 10-86 The that the thick filaments be compressed. The actual
colored ovals on each drawlng lndlcate reglons In whlch the ~nteractlon
slope of this decline and the length of the sarcomere at
between ftlaments IS crltlcal for determlnlng how much force can be gen-
erated by the sarcomere The equatlon accompanying each drawlng cal-
which no force can be produced cannot be predicted
culates the length of a s~nglesarcomere for that condltlon of overlap from the sliding-filament theory because these values
[Part B adapted from Gordon et al ,1966 ] would depend on the elastic modulus of the thick fila-
ments and on how many cross-bridges are generating
force.
Z disk. In a sarcomere that has been stretched just to
the point where the filaments fail to overlap, the ant How will changing the length of the filaments affect
the shape of the length-tenszon curve? Thin filaments
would have to cross one half of the thickness of a Z disk
(0.025 pm), crawl along 1 actin thin filament (1.0 pm), in the muscles of mammals are about 1.2 pm long,
step down to a thick filament and traverse it (1.6 pm), (i.e., about 0.2 pm longer than thin filaments in frogs).
step up to a thin filament on the other side of the sar- From this value and using the calculations shown in
comere and traverse it (1.0 pm), and finally cover the Figure 10-9, we would predict that the plateau of the
rest of the distance to the center of the second Z disk sarcomere length-tension graph of mammalian muscle
(0.025 pm). The total distance traveled would be would occur at sarcomere lengths between 2.45 and
3.65 pm (Figure 10-9B, condition 1). 2.65 pm and that force would fall to zero at 4.05 pm.
The length of the thick filaments can also affect the
Why is there a plateau in maximal force between 2.05 properties of the length-tension curve. In all verte-
and 2.25 pm? When the sarcomere is 2.25 pm long, the brates, the thick filaments are about 1.60 pm long, but
ends of the thin filaments line up with the beginning of they are considerably longer in some invertebrates.
the bare zone on the thick filaments (where there are Not only will the longer thick filaments change the
no cross-bridges). Thus all the cross-bridges on the shape of the sarcomere length-tension relationship, but
thick filaments are optimally aligned to interact with they will also change the absolute force. Longer thick
MUSCLES A N D A N I M A L M O V E M E N T 359
......................................
filaments can have more cross-bridges working in The observation that ATP is required for the dissociation
parallel and, hence, can generate higher forces (see of actomyosin (and for cross-bridge detachment) explains
Spotlight 10-1). a phenomenon well-known to readers of detective novels.
Following death, a human or other animal gradually
In the experiments that tested these predictions, it was becomes rigid and will hold the same position for hours
crucial that the length measurements be made on small o r even for days. This condition, called rigor mortis,
groups of sarcomeres located near the center of the muscle differs from muscle contraction, because in rigor mortis
fiber and that the sarcomeres behave uniformly. Measure- the muscles d o not shorten. Instead, they simply main-
ments made earlier, with less precise techniques, yielded tain the same length for a long time. This rigidity occurs
rounded curves because the thick and thin filaments in because when all of the ATP is used up after a cell dies,
many sarcomeres of a whole muscle-and, indeed, of myosin binds to actin and cannot detach, producing
a single fiber-varied in the amount that the thick and rigidity.
thin fibers overlapped at any given instant. A rounded When ATP binds to myosin, it is rapidly hydrolyzed to
curve would have failed to confirm the predictions of the ADP and PI, but the breakdown products unbind from
sliding-filament theory and have seriously misled muscle myosin only very slowly. Thus the rate of ATP hydrolysis
physiologists. by myosin is very slow, and the rate-limiting step is the
release of ADP and Pi from myosin. However, when actin
Cross-Bridges and the Production of Force binds to myosin, the release of ADP and Pi is greatly
Determining precisely how cross-bridges work is one of the speeded up, presumably through an allosteric change in
greatest challenges facing current researchers studying mus- the conformation of myosin. This actin-induced effect
cle function. According to recent versions of the sliding-fil- greatly increases the rate at which myosin can hydro-
ament theory, the force driving muscle contraction arises lyze ATP:
when several different sites on the myosin head bind se-
quentially to sites on the actin filaments. All bonds between
the head and the actin filaments are then broken, freeing the
M-ATP - M-ADP-PI M + ADP + PI
head for another cycle of sequential binding at a site farther
along the actin filament. We will now consider these
M-ADP-PI + A * AM + ADP + PI
processes in detail. Because the binding of actin to the myosin-ADP-PIcom-
plex releases energy, formation of actomyosin (AM) is ki-
Cross-bridge chemistry netically favored. Combining these reactions produces a cy-
Myosin cross-bridges must attach to binding sites on actin cle of binding and unbinding (Figure 10-10).The net effect
filaments in order to generate force, but cross-bridges must of one turn of the cycle is to split one molecule of ATP into
also be able to detach. If cross-bridges never detached from ADP + P, ,liberating energy.
actin, a muscle could never relax. In addition, if cross-
bridges could not detach, a muscle would be unable to
shorten; attached cross-bridges would prevent filaments
from sliding past one another, locking the muscle at one
length. As a result, the cross-bridges must attach and detach
ATP 7 Myos~n ATP
hydrolysis
Myos~n-ADP-P,
/
Myosin
Such a system poses a challenge to biochemists: How
can myosin first attach to actin, so force can be generated,
/Energyrelease
and then detach, so the filaments can continue sliding or the
muscle can relax? The first hints about the chemistry of the rr
Actornyosin
interaction between myosin cross-bridges and actin fila- + ADP + P, 1
ments came from studies begun several decades ago on
crude and purified extracts of muscle. Several interesting
physical properties were exhibited by semipurified solu-
tions of actin and myosin, which were extracted with con-
centrated salt solutions from freshly minced rabbit muscle
and then precipitated with ammonium chloride.
When actin (A) and myosin ( M ) are mixed in the ab-
sence of ATP, they form a stable complex called actomyosin Figure 10-10 In the presence of ATP, myosin and actin filaments asso-
(AM).However, the addition of ATP to the solution causes ciate and dissociate in a cycle. ATP binds to actomyosin, splitting the mol-
ecule into actin and myosin. The myosin then acts as an ATPase, hy-
rapid dissociation of the complex into actin and myosin-
drolyzing ATP, but the release of the products ADP and PI is slow unless
ATP: actin rebinds to the myosin, increasing the rate of release. The net result
of each turn of the cycle is the hydrolysis of one molecule of ATP, liberat-
AM + ATP = A + M-ATP ing energy, which can b e used to generate force.
SPOTLIGHT 10-1 gether. The force exerted by a thin filament on a thick filament is
equal and opposite to the force exerted by the thick filament on
PARALLEL AND SERIES the thin filament. The opposite polarity of the cross-bridges at
the two ends of a thick filament means that a thin filament on one
ARRANGEMENTS: THE end of a sarcomere exerts a force onto that thick filament that is
just balanced by the force of a matching thin filament on the
GEOMETRY OF MUSCLE
other side. Hence the net force exerted on a thick filament by the
surrounding thin filaments is zero, and the thick filament stays in
Muscle has a highly organized, crystal-like geometry-all the the center of the sarcomere (part A of diagram). For example, if
way from the structure of the filaments to the organization of the cross-bridges on the right side of a sarcomere are generat-
whole muscles. In this arrangement, some structural compo- ing a force of 100, then the cross-bridges on the left side must
nents are arranged in parallel with one another, while others are also be generating a force of 100, so the sum of the force on the
in series. These arrangements strongly affect the mechanics of thick filament is zero and the thick filament will stay in the middle
muscle contraction. of the sarcomere. However, if you were to attach a force trans-
Cross-bridges on one end of a thick filament are arranged in ducer to one of the Z disks and measure the force generated by
parallel with one another, but cross-bridges formed by the two the cross-bridges, you would measure a force of 100, that is, the
ends of a filament oppose each other. Every cross-bridge ex- force generated by the bridges in one-half of the sarcomere.
tends from a thick filament to a thin filament independent of all What would happen if this polarity wasn't built into the thick
other cross-bridges. Because of this arrangement, the forces filament? If all of the cross-bridges along a thick filament were
produced by the cross-bridges along one thick filament are ad- lined up in the same direction, the thin filaments v~ouldexert a
ditive, like the force produced by each person in a tug of war or force in only one direction, and the thick filament would travel
a current that moves through parallel resistors in a circuit. The along the thin filaments toward one of the Z disks (part B of dia-
force in one direction generated by a thick filament is equal to gram).There would be a unidirectional net force on the thickfil-
the force per cross-bridge times the number of cross-bridges on ament (to the right in the diagram), and the thick filament would
one half of the thick filament. What about the cross-bridges on travel toward the right Z disk with unpredictable results. In this
the other half of the thick filament? situation the sarcomere would not be able to generate force by
Hugh Huxley was the first to observe that the myosin shortening.
monomers making up one half of a thick filament are assembled Some invertebrates have long thick filaments with many
with their heads all pointed toward one Z disk, whereas those cross-bridges acting in parallel and the potential for generating
that make up the other half are oriented with their heads toward greaterforce. However, this potential depends on the ratio of the
the other Z disk. This polarized configuration is crucial for the ef- number of cross-bridges to the total length of the filament,
fective generation of force. Each set of cross-bridgesexerts force which in turn depends on the fraction of the length of each thick
on the thin filament directed toward the center of the sarcomere, filament that is populated by cross-bridges (i.e., excluding the
so the force produced by the cross-bridges pull the Z disks to- bare zones).
change in tension. Once it was stretched, the link would goes to the second, third, and fourth sites in succes-
transmit its tension smoothly to the thick filament, gener- sion, forming successively stronger myosin-actin in-
ating force to shorten the sarcomere. One major piece of teractions with lower energy states.
evidence supporting this hypothesis is the observation that 2. This sequential binding causes the myosin head to
the series elasticity of a muscle fiber is proportional to the rock or rotate, pulling on the link that connects the
-amount of overlap between the thick and thin filaments; myosin head to the thick filament. The elasticity of the
hence it is proportional to the number of attached cross- link allows the steplike rocking of the head to progress
bridges. Moreover, sudden small decreases in fiber length without sudden large changes in tension.
are accompanied by very rapid recovery of tension, which 3. Tension in the link is transmitted to the myosin
presumably results from rotation of the cross-bridge heads filament.
into their more stable positions of interaction with actin 4. When the rotation of the head is complete, the myosin
sites (i.e., from the MI to the M,sites). head dissociates from the actin filament and rotates
Not all details of cross-bridge function have yet been back to its relaxed position.
rigorously established. However, our present understand-
ing of the sequence of events in cross-bridge function is The myosin head detaches from actin only if Mg2+-ATP
summarized as follows (Figure 10-llC): binds to the ATPase site of the head region. The ATP is then
hydrolyzed, which is accompanied by a conformational
1. The head of each cross-bridge attaches to an actin fil- change in the myosin head, leaving the head in an energized
ament by the first of a sequence of stable sites. It then state, ready to rebind to a site on the actin that is a little
362 PHYSIOLOGICAL PROCESSES
.......................................
A B
A c t i n filament Actin filament
\
Cross-bridge link
1 Myosin
head
Y . ' l '
I
M y o s i n filament
Myosin filament
Head rotation
\
Myosin head
attaches t o a c t i n
hydrolysis pT '
Head
detaches
Figure 10-11 Thick and thin filaments slide past one another driven by head. These sites, M, to M,, interact in sequence (left to right) with sites
changes in bonds between myosin cross-bridges and actin. (A) Sequence on the actln f~lament.The rotation of the head produced in this manner
of events in the attachment of cross-bridges to actin filaments. In the re- causes the head to pull on the elastic cross-bridge Ihnk, stretching it. This
laxed state (top), myosin heads are not bound to actin. Myosin heads at- tension is transferredto the actin filament, pulling it toward the left in this
tach to actin (rniddle)and through the formation of a series of bonds, the diagram and causing it to slide past the myosin filament. (C) Summary
heads rotate, pulling on the actin filament (bottom)and causing itto slide of cross-bridge cycle. Note that the myosin head detaches only if ATP
past the myosin filament. Although the cross-bridges are shown acting in binds. [PartsA and B adapted from Huxley and Simmons, 1971; part C
unison, they actually act asynchronously. (B) Modeling of cross-bridge adapted from Keynes and Aidley, 1981.]
function suggests that there are four separate binding sites on the myosin
farther along the thin filament. This cycle is repeated over tions have been categorized based on what happens to the
and over, and the filaments slide past one another in small length of active muscles. In an isometric (i.e., "same
incremental steps of attachment, rotation, and detachment length") contraction, the length of a muscle is held fixed,
of the many cross-bridges on each thick filament. preventing it from shortening (Figure 10-12A). The previ-
ous discussion of the length-tension relationship for a sar-
comere was based exclusively on isometric contractions.
MECHANICS OF MUSCLE
Note that although no external shortening is permitted
CONTRACTION
during an isometric contraction, there can be a very small
Many of the mechanical properties of contracting muscle amount of internal shortening (about 1%), which occurs
were elucidated before 1950, when the mechanism of when intracellular and extracellular elastic components-
contraction was not yet understood. It is useful to con- such as cross-bridge links and connective tissue that is in
sider these classic findings and to attempt to explain them series with the muscle fibers-are stretched. In an isotonic
in terms of our current understanding of cross-bridge (i.e., "same tension") contraction, the muscle shortens as
behavior. force is generated (Figure 10-12B). Such contractions are
The term contraction refers to the activation of mus- the familiar ones that produce most movements. Contrac-
cles and the resultant generation of force. Muscle contrac- tion also may occur even when a muscle is lengthened by
Relaxed Active
Tendon
lsometrrc Muscle
Tension
Tendon
Isotonic
Muscle
- Tension Tens~on
Figure 10-12 Muscles can contract isometrically or isotonically. In iso- cause the heavy weight of the car would prevent your arm muscles from
metric contraction, the length of the muscle is held flxed, either by an ex- shortening. In isotonlc contraction, the muscle is allowed to shorten dur-
perimenter or by the physical situation. For example, if you tried to pick ing the tlme when tension is being generated. Isotonic contractions of
up your car with your left arm, the contraction would be isometric, be- muscles move joints when we walk, run, and produce other movements.
externally applied force while it is generating force, for ex- contract, and when the force generated by the muscle be-
ample if a heavy weight is suddenly applied to a contract- comes equal to the force exerted on the weight by gravity,
ing muscle. the muscle begins to shorten at a constant velocity (i.e., the
muscle begins to contract isotonically).
Relation between Force and Shortening Velocity In the example depicted in Figure 10-13B, the maximal
For animals to move, muscles must shorten, so the relation isometric tension that the muscle can produce is 100 g; that
between the production of force and the rate at which a is, if the muscle contracts against a load of 100 g or more,
muscle shortens (the so-called force-velocity curve) is cru- it cannot shorten. If the load is less than 100 g, the muscle
cial for understanding the design of a muscular system. His- will shorten slowly. If a 50-g weight is attached, the muscle
torically, to measure the force-velocity relation a muscle will shorten at a slow velocity; if lighter weights are at-
was attached to a lever with a weight attached to the other tached, the muscle shortens faster. When it shortens against
side of the fulcrum (Figure 10-13A).In more recent exper- no weight at all, it shortens at its fastest speed, which is the
iments, a motor controlled by a feedback circuit (a servo- maximal velocity of shortening, Vmax.Plotting the force
motor) replaces the weight, providing finer control. The generated against the shortening velocity generates a hy-
system is arranged so there is a limit on how much the perbolic curve that Archibald V. Hill, an important pioneer
weight, or the servomotor, can stretch the muscle. In this in muscle physiology, described in the 1930s by the fol-
setup, when the muscle is electrically stimulated, it starts to lowing equation:
A B
Fulcrum
Initial length
Lever
Figure 10-13 The force against which a muscle works and the velocity
Weight c at which it shortens are reciprocally related. (A) Typical setup for measur-
ing the relat~onshlpof force and velocity of a muscle. The muscle works
Stimulator against the welght that 1s hung on the other side of the fulcrum of the
lever. When the stimulated muscle generates a larger force than the
Force
weight, it shortens and pulls down on the lever. A servomotor system
I
trraniducer Time
could be used in this kind of experiment to provide finer control of the ini-
tial muscle length and the load. (B)The rate at whlch the muscle contracts
as it works against four different loads: 100 g, 50 g, 20 g, and 0 g. When
the load is smaller, so less force must be generated, the muscle contracts
more rapidly. The maxlmal isometric tension of this muscle is 100 g, so
when it works against the 100 g weight, it cannot shorten At the start of
the experiment, the length of the muscle would be set to optimize the
overlap between thick and thin filaments in the sarcomeres. (C) Force-
velocity curve based on the data shown in part B. At the maxlmal force of
100 g, the velocity of shortening is zero; that is, contraction is isometric.
Veloc~ty I Velocity I (D) Power-velocity curve calculated by multiplying the force and the ve-
lsometr~c Load locity for each data point in part C. The power is zero if either the force or
contract~on is zero the velocity equals zero.
under each condition. As shown in Figure 10-13D, the
power generated is maximal at intermediate velocities.
Power falls to zero if either the velocity of shortening or the
where V is the velocity of shortening; P, the force (or load); force is zero.
Po, the maximal isometric tension of that muscle; 6, a con- As we will see later in this chapter, it is useful to de-
stant with dimensions of velocity; and a, a constant with di- scribe force production or power production as V/VmaX,
mensions of force (Figure 10-13C). where V is the velocity of shortening in a particular condi-
Equation 10-1 implies that as the load increases, the tion and Vma,is the maximal velocity of shortening. Power
shortening velocity decreases. You probably are familiar production by the muscle shown in Figure 10-13 is maxi-
with this principle from personal experience; you can lift a mal at a V/Vmaxof about 0.15-0.4; this relation appears to
feather from a table much more rapidly than you can lift hold for all muscles, no matter how fast their V, is.
a heavy weight. Notice that the decline in force with in-
Effect of Cross-Bridges on Force-Velocity Relation
creased velocity does not reflect a change in myofilament
overlap. On the contrary, these experiments are purposely From the force-velocity curve described in the last section,
performed at the plateau of the length-tension relationship, we know that the force generated by a muscle drops as its
so the number of cross-bridges that can interact with actin speed of shortening increases. Recall that this relation is not
remains high and unchanged during shortening. caused by a change in the amount of overlap between thin
The relation between power and velocity is as impor- and thick filaments-rather, it is observed at the maximal
tant as the relation between force and velocity. For a fish to overlap. From our previous discussion of the role played by
swim or a frog to jump, its muscles must generate mechan- cross-bridges in isometric contraction, this fall in force with
ical power. The mechanical work performed by a muscle is increased velocity could result if fewer cross-bridges are at-
the product of force times length change (AL).Mechanical tached during rapid shortening, if each of the cross-bridges
power is given by that are attached generate a smaller force, or both. Andrew
Huxley's 1957 model of cross-bridge kinetics, although it
work (force)(AL) has been superseded in some fine details, still provides the
power = -- - = (force)(velocity)
time time basic principles for understanding the overall mechanics
and energetics of muscle contraction.
Hence, multiplying the force generated by the muscle by the According to Huxley's model, cross-bridges are con-
velocity at which it shortens yields the power production sidered to be elastic structures that generate zero force
SPOTLIGHT 10-2 made to contract and relax) if they are placed ~nappropr~atecon-
dltlons In such f~bers,an lnvestlgator can control the composl-
SKINNED MUSCLE FIBERS t ~ o nof the lntracellular f ~ u ~ without
ds any Interference from reg-
ulatory mechanisms that are normally present in an intact muscle
One of the early advances that contributed significantly to mus- fiber.
cle-cell physiology occurred when Albert Szent-Gyorgyi devel- Another method of extracting some substances from cells,
oped a procedure for isolating muscle fibers in which the intra- while leaving the insoluble proteins intact, employs nonionicde-
cellular structure remains intact, but the membrane no longer tergents, such as the Triton X series. These agents, which are
preventsfree exchange of materials between the cytoplasm and used at about O°C, rapidly solubilize the lipid components of the
the extracellular solution. This kind of preparation is called cell membrane, allowing soluble metabolites to diffuse out of
a "skinned" muscle fiber because its outer membrane has been the cell and substances in the extracellular medium to diffuse
entirely removed or rendered so leaky that it is functionally rapidly into the cell. Fibers treated in this way are called "chem-
absent. ically skinned muscle fibers." This process requires only minutes,
In Szent-Gyorgyi's procedure, muscle fibers are soaked for rather than days or weeks as is the case of glycerin extraction,
several days or weeks at a temperature below 0°C in a solution thus saving much time.
made up of equal parts of glycerin and water. Under these con- A final way of producing skinned fibers is to actually dissect
ditions the cell membrane becomes disrupted, and all soluble away the cell membrane using fine forceps. This process, which
substances in the myoplasm are leached out, leaving intact the resembles removing the casing from a link sausage, requires
insoluble molecules that make up the contractile machinery. The great manual dexterity. With practice, however, a person can
glycerin in the solution prevents the formation of ice crystals, prepare fibers that remain structurally intact by this method.
which could break up the structural organization of the fibers, Regardless of which method is used to prepare skinned
and it also helps to solubilize the membranes.Storing the tissue muscle fibers, all ofthem allow experimentersto manipulate the
at low temperature preservesthe enzymes, but slows down cata- chemical environment surrounding the contractile machinery of
bolic processes that would cause the cells to digest themselves. muscle fibers, expanding the opportunities to understand the
These glycerin-extracted muscle fibers can be reactivated (i.e., molecular basis of contraction.
MUSCLES A N D A N I M A L MOVEMENT 365
......................................
when they are at equilibrium. This behavior is similar to a A
Equilibrium
F
piece of spring steel projecting out from a surface. If the Actin, position Z diskl .
piece of steel is deformed by bending it, a restoring force is
created to return it to its original position. Similarly, when
a cross-bridge is bent toward or away from the Z disk, a
restoring force is created that tends to bring it back to its Position
original position; the magnitude of this force is propor-
tional to the displacement of the cross-bridge from the equi-
librium position (Figure 10-14A). If a cross-bridge bent
toward the Z disk were attached to a thin filament, the
restoring force would pull the Z disk toward the center
of the sarcomere; this force is considered to be in the
"positive" direction. By contrast, if a cross-bridge bent
away from the Z disk were attached to a thin filament, Thin filament
the restoring force would push the Z disk away from the
center of the sarcomere; this is viewed as a "negative"
force.
Figure 10-14B illustrates how the forces generated by
cross-bridge displacement cause movement of a thin fila-
ment. When the cross-bridge is at the equilibrium position
(O), the force, Fo, is zero; when the cross-bridge is bent to-
Thick filament
Ca2+ removed
Figure 10-15 Free calcium ions regulate the state of muscle contraction. varies with the concentration of Ca2+in the surrounding medium. Force
(A) Glycerin-extracted muscle fibers generate tension when they are ex- increases with increasing Ca2+concentration , up to some maximum
posed to Ca2+and Mg-ATP. They relax when Ca2+is removed, even if value. [Adaptedfrom Hellam and Podolsky, 1967.1
Mg-ATP is still present.(B) The force generated by a skinned muscle fiber
As discussed earlier, the ATPase activity of myosin heads caused all the attached cross-bridges to be frozen in place.
increases dramatically when the heads bind to actin. Be- Adding Mg-ATP back to the muscle that was in rigor and
cause Ca2+increases the binding of myosin heads, it would bathed in Ca2+-freesolution caused it to relax. Thus, both
be expected to increase the ATPase activity of myosin. In- ATP and Ca2+must be present if the thick and thin filaments
deed, the ATPase activity of skinned fibers has been shown are to interact effectively to produce tension.
to increase with the Ca2+concentration of the surrounding The role of Ca2+in regulating the actin-myosin inter-
solution (Figure 10-17A). The normal cycling of muscle action via troponin and tropomyosin applies to vertebrate
contraction and relaxation depends on the presence of both skeletal and cardiac muscle. The role of Ca2+ differs in
ATP and Ca2+in the cytosol of muscle fibers. This is demon- most other muscles. At least two other calcium-dependent
strated by the data in Figure 10-17B. In this experiment, regulatory mechanisms controlling actin-myosin interac-
when a glycerin-extractedmuscle fiber was initially exposed tions exist. In most invertebrate striated muscles, calcium
to Ca2+in the absence of Mg-ATP, no tension was gener- initiates contraction by binding to the myosin light chains
ated. When Mg-ATP was added, tension developed, and of the cross-bridge heads. Contraction of vertebrate smooth
that tension was then maintained even when Mg-ATP was muscle and of nonmuscle actomyosin depends on a cal-
removed; that is, rigor mortis set in. Once the muscle was in cium-dependent phosphorylation of the myosin head, as
rigor, removal of Ca2+had no effect because the lack of ATP described in the last section of this chapter.
A
Figure 10-16 Tropon~nand tropomyosln reg-
ulate blndlng between myosln cross-br~dges
and actln thin f~laments (A) Three-d~men-
s~onalmodel dep~ctlngthe assoclatlon of
tropomyosln (TM)and the troponln complex,
contalnlng subun~tsC, I and T, w ~ t hthe actln
B filament (B) Mechan~smof Ca2+med~ated
regulat~onof act~n-myos~n ~nteract~on When
the concentration of Ca2+1s low (left), the tro-
ponln complex b~nds with actln and
tropomyosln, sterically preventing myosln
cross-br~dges from blndlng to actin If the con-
centratlon of cytoplasm~cCa2+increases tro-
ponln C b~ndsCa2+ chang~ngthe subun~t
affln~tiesand causlng the tropomyosln mole-
cule t o move away from the myos~n-b~nd~ng
slte on actin Cross-br~dgescan then b ~ n d
cycl~cally,and the th~ckand t h ~ n
f~lamentscan
sl~depast one another unt~lCa2+IS removed
from the tropon~ncomplex [Part A adapted
from Phill~pset al , 1986, part B adapted from
Ebashi et al , 19801
368 P H Y S I O L O G I C A L PROCESSES
...............................................................................
Caz+ present
I - Ca2+ absent i
I
,U I Time
t
Mg-ATP
Time (min)
t
Ca2+
Latent
period
Sarcomere Z disk
A
- - --
Figure 10-20 When frog muscles are stimulated by an extracellular ml- st~mulat~ng
-Sarcomere Z disk
Transverse ---
Sarcoplasmic-
Longitudinal-
t Br~ef
~~~~~~d
'Igh"'
cence, the membrane potent~al,and the pro-
ducton of tens~onby the f~berare recorded
(B) When the muscle f~ber1s st~mulated,an
strmulus AP is propagated along the surface mem-
brane and recorded by the recording mlcro-
electrode A short t ~ m elater, the fluorescence
s~gnalfrom the calc~um-sens~t~ve dye ~ n s ~ d e
the f~ber~nd~cates that the Ca2+concentra-
tion w ~ t h ~the
n f~berhas Increased, and even
later, the tension transducer measures the
produalon of tens~onby the f~berNot~ce that
the tenslon beglns to rlse only after the AP is
over and the lntracellular Ca2* concentratlon
1s decllnlng [Part B courtesy of S M Baylor]
M period
MUSCLES A N D A N I M A L MOVEMENT 373
...............................................................................
revealed three associated tubes or sacks. Two of the sacks voltage-sensitive. Because the ryanodine receptors extend
are always large, and they are located on either side of a most of the way across the cleft, a direct mechanical link-
much smaller tube or sack. We now know that the two age between ryanodine receptors and dihydropyridine re-
sacks are two terminal cisternae of the sarcoplasmic retic- ceptors was proposed (Figure 10-25). So the model pro-
ulum and the smaller central tube is a T tubule. How can poses that when the T tubule depolarizes, the voltage-
an AP in a T tubule be conveyed to the sarcoplasmic retic- sensitive dihydropyridine receptor undergoes a conforma-
ulum, causing it to release Ca2+?Although the details are tional change and either mechanically dislodges the ryan-
not yet completely understood, the general mechanism is odine receptor from calcium-selective channels in the SR
now known. membrane or forces the ryanodine receptor into a confor-
mational change that opens the calcium channels.
Membrane receptors in triads In 1970, electron micro- Interestingly, only about half of the ryanodine receptors
scopy experiments by Clara Franzini-Armstrong revealed are associated directly with voltage-sensitive dihydropyri-
electron-dense molecules in the part of the SR membrane dine receptors in the T-tubule membrane. This finding sug-
that lies adjacent to the T tubule. She called these molecules gests that if there is a direct mechanical linkage between T
"feet;" they have more recently been named the ryanodine tubules and the sarcoplasmic reticulum, only about half of
receptor, because they bind the drug ryanodine. Knox the ryanodine receptors participate. It has been proposed
Chandler and his colleagues then proposed that these pro- that the other, unlinked ryanodine receptors are activated by
teins represent Ca2+channels and incorporated them into the increase in myoplasmic free Ca2' resulting from open-
the "plunger model" for release of Ca2+ from the sar- ing of the mechanically linked channels. Activation of these
coplasmic reticulum (Figure 10-24). In this model, depo- unlinked ryanodine receptors would in turn open more
larization of the T tubule causes a plug to be removed from Ca2+channels in the membrane of the sarcoplasmic reticu-
the Ca2+channels in the SR membrane, allowing Ca2+to lum. Such a mechanism, called Ca2+-inducedCa2+release,
escape into the myoplasm driven down its steep electro- has been found in other tissues including the excitation-con-
chemical gradient. When the T-tubule membrane repolar- traction coupling mechanism in cardiac muscle.
izes, the plug is replaced, preventing further Ca2+release.
The plunger model proposed an explanation of how Time course of calcium release and reuptake The ability to
the T-tubule membrane and the SR membrane might be measure rapid changes in the myoplasmic Ca2+concentra-
coupled, but it failed to identify what caused the plunger to tion, combined with the information about calcium bind-
move in response to depolarization of the T-tubule mem- ing by troponin and the kinetics of the calcium pumps in
brane. Additional electron microscopy studies revealed a the SR membrane, has permitted the Ca2' fluxes during
cluster of proteins in the T-tubule membrane directly across muscle contraction and relaxation to be modeled. Such
from each ryanodine receptor in the SR membrane. These modeling suggests that when T tubules become depolar-
T-tubule proteins, called dihydropyridine receptors, are ized, Ca2+ flows out of the sarcoplasmic reticulum for
@ Dihydropyridine
receptpr Lumen of
0
Figure 10-24 Depolarizationof theT-tubule membrane indiredly causes flow out of the SR lumen into the myoplasm.The free Ca2+binds to tro-
calcium channels in the sarcoplasmic reticulum to open. When the mem- ponin, revealing cross-bridgebinding sites on actin molecules. When the
brane of theT tubule is at rest (I), calcium channels in the SR membrane membrane potential returns to rest (3),the ryanodine receptors again
are blocked by the "feet" of ryanodine receptors. When the T-tubule block the calcium channels. Calclum pumps in the sarcoplasmic reticu-
membrane depolarizes (2), voltage-sensitive d~hydropyridinereceptors lum resequester Ca2+,shifting the equilibrium of Ca2+binding on tro-
convey the signal to the ryanodlne receptors, and the "plungers" block- ponin and concealing the cross-bridge binding sites. [Adapted from
ing calcium channels in the SR membrane are removed, allowing Ca2+to Berridge, 1993.1
374 PHYSIOLOGICAL PROCESSES
............................................
1. The surface membrane of the fiber is depolarized by an
AP or, in some muscles, by synaptic potentials. In an
animal, APs in skeletal muscle fibers are generated by
synaptic potentials, so neuronal input is required for
initiating contraction in skeletal muscle.
The AP is conducted deep into the muscle fiber along
the T tubules.
In response to depolarization of the T-tubule mem-
brane, voltage-sensitive dihydr~p~ridinereceptors in
the T-tubule membrane undergo a conformational
change that-through direct mechanical linkage to
ryanodine receptors in the SR membrane-causes
opening of Ca2+ channels in the SR membrane (see
Figure 10-24, steps 1and 2).
As Ca2+flows out from the lumen of the sarcoplasmic
reticulum, the free Ca2+ concentration of the myo-
plasm increases from a resting value of below lo-' M
to an active level of about M, or higher, within a
few milliseconds. The Ca2+channels in the SR mem-
brane then close.
Most of the Ca2+ that enters the myoplasm binds
rapidly to troponin, inducing a conformational change
in the troponin molecules. This conformational
change causes a change in the position of the tropo-
Figure 10-25 In an intact triad, several molecules contribute to the con- myosin molecule, eliminating steric hindrance and al-
trol of myoplasmic calcium. The voltage-sensitive dihydropyridine re- lowing myosin cross-bridges to bind to actin thin fila-
ceptor and the ryanodine receptor work together linking depolarization
ments (see Figure 10-168).
of the T tubule to opening of calcium-selective channels in the SR mem-
brane through which Ca2+movesfrom the SR lumen into the myoplasm. 6. Myosin cross-bridges attach to the actin filaments and
The feet of the ryanodine receptors extend into the gap between the T go through a series of binding steps that cause the
tubules and sarcoplasmic reticulum.A calcium ATPase (calcium pump) in
the SR membrane resequesters Ca2+from the myoplasm, and calse-
myosin head to rotate against the actin filaments,
questrin inside the SR binds Ca2+,reducing the concentration of free pulling on the cross-bridge link (see Figure 10-11A,B).
ionic Ca2+inside the SR. [Adapted from Block et al., 1988.1 This pulling produces force on, and in some cases ac-
tive sliding of, the thin filaments toward the center of
the sarcomere, causing the sarcomere to shorten by a
several milliseconds and then the calcium channels close.
small amount (see Figure 10-8A).
The mechanism for the closure is not fully understood.
Most of the Ca2+that leaves the sarcoplasrnic reticulum ATP binds to the ATPase site on the myosin head caus-
binds very quickly to troponin. The concentration of tro- ing the myosin head to detach from the thin filament.
ponin in muscle fibers is about 240 pM, which represents ATP is then hydrolyzed, and the energy of the hydro-
a large buffer for Ca2+ions. Thus, a very small amount of lysis is stored as a conformational change in the
the released Ca2+remains free in myoplasm, and it is only myosin molecule, which then reattaches to the next
this small amount of unbound Ca2+that is detected by a site along the actin filament as long as binding sites are
calcium-indicator dye in experiments such as the one still available, and the cycle of binding and unbinding
shown in Figure 10-23. Both during and following the re- is repeated (see Figure 10-11C). During a single con-
lease of Ca2+ from the sarcoplasmic reticulum, the free traction, each cross-bridge attaches, pulls, and de-
Ca2+in the myoplasm is pumped back into the SR lumen, taches many times as it progresses along the actin fil-
lowering the myoplasmic level of free Ca2+.As the concen- ament toward the Z disk.
tration of free Ca2+ in the myoplasm becomes very low, Finally, calcium pumps in the SR membrane actively
8.
Ca2+bound to troponin is released back into the myoplasm
transport Ca2+ from the myoplasm back into the
and then is subsequently pumped back into the sarcoplas- SR lumen (see Figure 10-24, step 3). As the concentra-
mic reticulum where it binds to calsequestrin.
tion of free Ca2+in the myoplasm drops, Ca2+bound
Contraction-Relaxation Cycle to troponin is released, allowing tropomyosin again
Starting with a relaxed skeletal muscle, the following se- to inhibit cross-bridge attachment, so the muscle re-
quence of events leads to contraction and then relaxation laxes. The muscle remains relaxed until the next
of a skeletal muscle fiber: depolarization.
MUSCLES A N D A N I M A L M O V E M E N T 375
...............................................................................
THE TRANSIENT PRODUCTION Series Elastic Component
OF FORCE A muscle can be modeled as a contractile element that
is arranged in parallel with one elastic component and
Up to now, we have considered the mechanics of myosin in series with another elastic component as depicted in
cross-bridges that have been activated maximally. As Figure 10-26A. (According to Hooke's law, the length
we've seen already, there is a delay, or latent period, of an object with ideal elasticity increases in propor-
be-tween the action potential in a muscle fiber and the gen- tion to the force applied.) The parallel elastic compo-
eration of force by the fiber (see Figures 10-18B and nent in this model represents the properties of the
10-23B). The latent period includes all of the time that is plasma membrane of the muscle fibers and connec-
required for initiation of an action potential in the muscle tive tissues that run in parallel with the muscle fibers.
fiber, propagation of the AP along the T tubules into the The series elastic component, also called the series
fiber, release of Ca2+from the sarcoplasmic reticulum and elastic elements, represents tendons, connective tis-
diffusion of the Ca2+ions to troponin molecules, binding sues that link muscle fibers to the tendons, and per-
of Ca2+ to troponin, activation of myosin cross-bridges haps the Z disks of the sarcomeres. An additional
and their binding to actin thin filaments, and generation of important constituent of the series elastic elements appears
force. The time required for all of these processes is short: to be the myosin cross-bridge links themselves, which ap-
The delay from the peak of the AP to the first sign of ten- pear to undergo some stretch in response to tension (see
sion in some muscles can be as short as 2 ms. Now we con- Figure 10-11B).Representing all of the elastic components
sider the mechanical properties of muscle fibers as they by only two components greatly simplifies the model,
become activated and generate tension and then relax in making it easier to manipulate mathematically while
the body. maintaining sufficient accuracy that the model can
component
Time (msec)
High-frequency APs by which individual twitches fuse can vary, reaching a max-
(tetanus) imal value in tetanus (Figure 10-28).
ENERGETICS OF MUSCLE
CONTRACTION
In muscle contraction two major processes require the ex-
penditure of energy. The most obvious one is the hydroly-
sis of ATP by myosin cross-bridges as they cyclically attach
to and detach from actin thin filaments (see Figure 10-10).
The other major process that uses energy is the pumping of
- Ca2+ released into the myoplasm during excitation-con-
Time
traction coupling back into the sarcoplasmic reticulum
Figure 10-28 Twitches fuse when the stimulating APs arrive in rapid suc-
against a Ca2+concentration gradient (see Figure 10-24,
cession. A single AP produces a single twitch. If a set of low-frequency
APs IS conducted along the muscle fiber, each successive twitch begins
step 3).
before the muscle has time to return to its relaxed condition following the Biochemical studies have determined that 2 molecules
previous twitch. At a maximum frequency, the twitches fuse with one an- of ATP are required to pump each Ca2+ion into the sar-
other, producing a long, strong contraction called tetanus. coplasmic reticulum. During a twitch, a certain amount of
Ca2+is released in a few milliseconds after the AP, and ex-
actly that amount of Ca2+must eventually be pumped back
centration of Ca2+remains high in the myoplasm, and the into the SR if the muscle fiber is to relax and remain
active state is prolonged. With the active state prolonged, healthy. During tetanus (just as in a twitch), the calcium
isometric tension continues to increase with time until the pumps immediately start to resequester the Ca2+released
tension produced by the internal shortening of the con- during- the first stimulus; however, they do not have time to
tractile components and the stretching of the series elastic remove all of it from the myoplasm, and each successive AP
component is just sufficient to cause cross-bridges to slip causes more Ca2+to be released. The buildup of Ca2+in the
and prevent further shortening of the contractile compo- myoplasm keeps the troponin saturated with Ca2+until the
nents. The muscle has then reached full tetanus tension. De- APs cease; at that point, the calcium pumps eventually can
pending on the repetition rate of muscle APs, the amount resequester all of the released Ca2+back into the SR lumen.
378 PHYSIOLOGICAL PROCESSES
.......................................
To maintain the condition of tetanus, ATP is steadily hy- amount of ATP hydrolyzed by the muscle is best done by
drolyzed by both the myosin ATPase and the calcium measuring either the drop in the concentration of creatine
pumps in the membrane of the SR. phosphate or the rise in the concentration of PI.
Beyond the technical issue of accurately measuring
ATP Usage by Myosin ATPase and Calcium Pumps the rate of ATP hydrolysis, the creatine phosphoki-
The relative consumption of ATP by myosin ATPase and nase reaction is extremely important for effective muscle
the calcium pumps in the SR membrane was determined in function. If a muscle runs out of ATP, it goes into rigor (see
experiments with tetanized frog muscle. In these experi- Figure 10-17B). It is, therefore, essential for survival that
ments, the muscle preparations were stretched to different the concentration of ATP in muscles be buffered. During
degrees producing different amounts of overlap between endurance activities, oxidative and anaerobic metabolism
thick and thin filaments. As the muscle is stretched more can generate ATP rapidly enough to maintain a sufficient
and more, fewer cross-bridges can interact with actin, re- ATP concentration to power muscle contraction. However,
ducing both the amount of force that can be produced and during high-intensity, short-duration activity (e.g., when an
the amount of ATP hydrolyzed by the myosin ATPase. (Re- animal sprints to run down prey or to avoid becoming
member that myosin can hydrolyze ATP on its own, but the prey), the concentration of ATP within muscles is main-
resulting ADP and PI are released much more slowly than tained constant by continuous rephosphorylation of ADP
they are when the myosin is bound to actin. As a result, if by the creatine phosphokinase reaction (Figure 10-29).
myosin cross-bridges have few available sites where they The concentration of creatine phosphate in muscle
can bind to actin, the ATPase activity of the filaments will fibers (20-40 mM) is many times larger than the reserve of
be low.) By contrast, stretching the muscle should have lit- ATP (about 5 m M in muscle fibers). As a result, an animal
tle or no effect on the rate at which Ca2+is released from can use the large reserve of high-energy phosphate in crea-
and resequestered by the sarcoplasmic reticulum, because
these processes are mediated by membrane proteins whose
activity is unrelated to the amount of myofilament overlap.
Thus, as the muscles become increasingly stretched, the to-
tal ATPase activity will decline, and at the length where the
myofilaments no longer overlap any ATPase activity mea-
sured will be due entirely to the calcium pumps.
Using this experimental approach, researchers found
that the calcium pumps accounted for about 25% -30% of
total ATPase activity during muscle contraction. It is gen-
erally assumed that this percentage is constant for all mus-
cles; that is, muscles with a higher maximal contraction ve-
locity also have faster calcium pumps in their SR. It is
possible, however, that in the very fast sound-producing
muscles discussed later in this chapter, pumping of Ca2+
may account for a larger fraction of the overall energy us-
age of the muscle.
TABLE 10-1
Properties of twitch (phasic) fibers in mammalian skeletal muscles
Slow oxidative Fast oxidative Fast glycolytic
Property (type 1) (type Ila) (type Ilb)
Length-tension relation
To examine the length-tension relation of muscles during
a jump, Lutz and Rome measured the length and changes
in length of the semimembranous muscle, a hip extensor.
The measurements were made both while an intact frog Moment arm
jumped and in an isolated limb whose position was
manipulated to match the shape of a jumping frog's leg
(Figure 10-32). By plotting the length changes versus hip
angle, they determined the moment arm of the muscle. Pelvis flexed
(From physics, the moment arm is the distance separating
a fixed fulcrum from a point at which a force is exerted that
will tend to rotate a mass around the fixed point, as illus-
trated in the inset in Figure 10-32.)In this case, the moment
arm-that is, the distance between the hip joint and the lo-
cation where the muscle attaches to the pelvis-is crucial
because it determines both the mechanical advantage of the
muscle and the length changes it must make to produce any
given change in the angle of the hip joint.
The length of sarcomeres in the hip flexor was deter-
mined when the hip was in the crouched position and the
extended, jumping position. During a jump, the sarcomere
length goes from 2.34 pm in the crouched position to 1.82 Pelvis extended
p m at the point of take-off. To determine where these
lengths fall in the sarcomere length-tension relation, they
were compared with a length-tension curve from a closely
related species of frog (Rana temporaria) as shown in Figure
10-33A. The measured lengths of the hip flexor sarcomeres
fell along the plateau of the sarcomere length-tension curve;
thus, as expected for a power muscle, the fibers of the hip
flexor operate very near their optimal power production
during a jump. It has been calculated that this muscle gen-
- Muscle
Figure 10-32 When the hip flexor muscle contracts, the hip joint rotates
erates at least 90% of its maximal tension throughout the around the attachment pont between the pelvis and the femur. (A) When
jump. In addition, because of more subtle properties, if the the frog is in a crouched position, the angle of the hip jo~nt(8,) is small
initial sarcomere length were either longer or shorter than and the extensor muscle (the semimembranosus muscle) IS relaxed. (B)
When the frog jumps, the angle of the hip joint increases (O,), because
the measured length, the muscle would do less work.
contractlor- ofthe muscle acts through the moment arm indicated on the
A number of factors must be matched to produce this diagram and pulls on the pelv~s.Inset: Schemat~cdiagram illustrat~ngthe
optimal behavior. The lengths of the myofilaments and the mechanical components contributing t o jumping. [Adapted from Lutz
number of sarcomeres per muscle fiber must combine so and Rome, 1996.1
that there is optimal overlap of the thick and thin filaments
when the frog is in the crouched position. In addition, given matched to allow the hip flexor muscle t o shorten at a
the change in the angle of the hip joint that occurs during VIV,,,,, appropriate for maximal generation of power.
jumping, the moment arm of the hip joint must allow the
muscle and its sarcomeres to undergo the appropriate State of activation
changes in length. Even if the hip flexor muscle begins to contract at the opti-
mal sarcomere length and shortens at an optimal rate al-
Value of VIV,, lowing maximal power to be generated, it also must be
The Vmaxof the hip flexor muscle is about 10 muscle maximally activated to generate maximal power. If the mus-
lengths per second, and it generates maximal power at 3.44 cle started to shorten before it became fully activated, it
muscle lengths per second (Figure 10-33B). The mean rate would generate a force lower than the maximum possible at
at which the muscle shortens during a jump is 3.43 muscle that velocity (i.e., the actual force would lie below the force-
lengths per second, that is, at a VIV,,, of 0.32, precisely the velocity curve) and the power would also be lower than the
rate at which the muscle produces maximal power. Thus maximum. As discussed earlier, the time it takes for activa-
the frog's muscles, joint configuration, and mass are all tion to occur depends on the rate at which Ca2+is released
Figure 10-33 The mechanics of the hip flexor muscle of a frog operate
optimally during a jump. (A) At the beginning of a jump, the sarcomeres
in the semimembranous muscle are 2.34 p m long and they shorten to
1.83 p m during the jump (red portion of curve). Even at the shortest sar-
comere length, the muscle still generates over 90% of its maximal ten-
sion. (B) At the velocity used during jumping, the muscle operates in the
portion of the power curve (red) in which at least 99% of maximal power
is generated. The velocity of shortening is expressed in terms of muscle-
lengths per second to take into account the difference in length en-
countered among muscles. [Adapted from Lutz and Rome, 1994.1
- 2 1 - i
O
-
Lag
Figure 10-34 Electrical actlvlty patterns and
length changes recorded from muscles In an
~ntactfrog can be Imposed upon Isolated
muscles to study the force generated (A)
E
-
E
\ -5
E
E
Electromyogram recorded from the seml-
membranous muscle of a frog durlng a Jump
g
C
4-
\ 0
C
4 (left), and the abstracted pattern of st~mulat~on
-al
\
2 that was ~mposedon an lsolated muscle
4 4 (rlght) (B) Rate atwh~chthe muscle shortened
6- 6
b - In an Intact frog durlng jumplng and the
change In length Imposed on the Isolated
muscle whlle ~twas belng electr~callystlmu-
8 5 b 215 20 5; l b 0 1L5
8
lated as shown ~npart A (C)Force produced
by the Isolated muscle (solld Ihne) durlng the
T ~ m e(ms) experimental manlpulatlon mlrnlcklng the
!:;;
stlmulatlon and shorten~ngrate observed dur-
Figure 10-36 Swimming movements and the escape response in fish dif- lated from the shape of the fish at each time-point in a high speed
fer greatly in magnitude and time course, reflecting differences in motion picture film of the behavior. The shape of the fish body at
changes of sarcomere length. Shown here are the calculated changes in selected time points is indicated in figures below the graph. Type I (slow
sarcomere length wlthln the red muscles located on one slde of the an- oxidative) muscles are active during steady swimming. In contrast, type
terior, midbody, and posterior of a carp engaged in two activities: (A) Ilb (fast glycolflic) muscles produce the escape response (see Figure
.
swimmlng steadily at 25 cm s-' or (B) escape response eliclted by a loud 10-37). [Part A adapted from Rome et al., 1990a; part B adapted from
sound. The changes in sarcomere length of the red muscle were calcu- Rome et al., 1988.1
based upon measurements of sarcomere length made in fish comere length-tension curve for frog muscle provides a
' that had been frozen into the shapes assumed by living fish good approximation of the same relation in carp. Com-
as they perform different behaviors. These measurements parison of the sarcomere lengths measured in swimming
indicate that the sarcomeres in the red muscles of slowly carp with the frog length-tension relation shows that in
swimming fishes repeatedly vary in length between 1.89 swimming the red muscles generate at least 96% of their
p m and 2.25 pm, centered around a sarcomere length of maximal force (Figure 10-37A, left).
2.07 pm (see Figure 10-36A).These values then need to be In the escape response, the fish moves rapidly, and its
compared with the length-tension curve for fish sarcomeres body curves dramatically. Recall that the red muscles in
to determine if the thick and thin filaments maintain opti- carp run parallel to the long axis of the fish, whereas the
mal overlap while the sarcomeres undergo these changes in white muscles run helically (see Figure 10-35). Given the
length. Electron microscopic examination of red and white anatomic arrangement of the red muscles, the sarcomeres
muscles from carp reveal that the lengths of myofilaments would have to shorten to 1.4 pm, a length at which force is
in fish muscles are nearly identical with the lengths of these low (see Figure 10-36B),to produce the escape response. In
filaments in frog muscle. This finding indicates that the sar- contrast, the sarcomeres in white muscles need shorten only
A Steady swimming
"hVm
0.2
0.0
0
Red muscle
5
,
10 15
Sarcomere length (pm) Contraction velocity (muscle lengths. s-1)
B Escape response
Figure 10-37 The properties of red and white muscles of fish make them (B) Because of the~ranatomic arrangement, white fibers can produce the
optimally suited for different kinds of activities (A) The changes in the sar- escape response at a more favorable region of the sarcomere length-
comere length of red muscles during contraction coincide with the tension curve than can red fibers (left). In addition, the high V,, of white
plateau of the sarcomere length-tension curve (left) The colored bar in- fibers allows them to generate power when they are shortening very
dicates the lengths of red muscle sarcomeres during slow, steady swim- rapidly (right). Indeed, during the escape response the V/VmaXfor white
ming. In addition, the contraction velocities of red muscles during swim- muscles 1s 0 38, prec~selyat the peak of the~rpower curve. [Adapted from
ming correspond to values of VIV,,,,, between 0.17 and 0.36, near the Rome and Sosn~ck~, 1991 ]
value at which red f~bersproduce maximal power (right; colored region).
to about 1.75 p m during this behavior. In other words, the ments and the anatomic arrangement of the muscle fiber
mechanical advantage conferred by the anatomy of the types combine to allow this optimization.
white muscles allows them to produce a given change in the
curvature of the spine with much less sarcomere shortening Value of V/V,,
than would be required in red muscles. Thus, white mus- In addition to their different anatomic arrangements, the red
cles are much better suited to producing the escape re- and white muscles of a carp have different values of Vmax.
sponse and generate about 85% of their maximal force The V, of carp red muscle is 4.65 muscle lengths per sec-
during this behavior (see Figure 10-37B, left). When white ond, whereas the V,,,,, of carp white muscle is 12.8 muscle
muscle is used in less extreme movements (e.g., when a fish lengths per second, about 2.5 times higher. During steady
is swimming rapidly), the curvature of the backbone is not swimming the red muscle shortens at a V/V, of 0.17-0.36,
nearly as extreme, the sarcomeres shorten less, and the which is near the value at which maximal power is generated
muscles generate nearly maximal force. (Figure 10-37A, right). At higher swimming speeds (higher
Because the fish uses different muscles to produce dif- values of VIV,,,), a fish needs to generate greater mechani-
ferent movements, the myofilament overlap (sarcomere cal power, but at these higher values the mechanical power
length) is never far from its optimal level, even in the most output of the red muscle actually declines. In order to swim
extreme movements. The lengths of the thick and thin fila- faster, a fish must activate white muscles as well.
In contrast to steady swimming, the escape response de- side of the fish shorten, they will be most efficient in chang-
pends on activity in the white muscles. To power the escape ing the shape of the fish-allowing it to push against the
response, the red muscles would have to shorten at 20 mus- water-if the muscles on the other side of the body are re-
cle lengths per second-four times faster than their Vmax. laxed. If muscles on both sides of the fish contracted max-
White muscle in the anatomic orientation of the red mus- imally and simultaneously, the fish would be rigid and
cles would also be unable to power the escape response, be- straight. Thus it is essential for each muscle to relax after
cause the Vmaxof these muscles is only about 13 muscle it shortens, so that it offers no resistance when the con-
lengths per second. However, the helical arrangement of the
- -
tralateral muscle becomes active.
white muscles allows them to produce the escape response To better understand how the kinetics of activation and
when they shorten at only about five muscle lengths per sec- relaxation affects the generation of power during cyclical
ond, which corresponds to a VIV,,, of about 0.38, the muscle contractions, Robert Josephson has applied the
value at which the carp's white muscles produce the most "workloop" technique to muscles. In this approach, mus-
power (Figure 10-37B, right). cles are driven by a servomotor system through the cyclical
Perhaps a fish would be better off with only white mus- changes in length that occur during locomotion, and the in-
cles. The white muscles could certainly power slow swim- vestigator delivers a stimulus to the muscle at a particular
ming. However, the high Vmaxof white muscles would time in the cycle. In this kind of experiment, the timing of
mean that the VIV,,,,, during slow swimming would be so the stimulus, the duration of the stimulus, the intrinsic ac-
low (0.01-0.03) that they would be extremely inefficient. tivation and relaxation rate of the muscle, and the value of
Red muscles also can produce adequate power to generate Vmaxfor the muscle interact to determine how much power
slow swimming, and they do it much more efficiently than the muscle generates.
white muscles could. Thus, the anatomic arrangement and A useful way for quantifying these potentially complex
the V,, of the two kinds of muscle suits them to the par- interactions is to measure the amount of net work ( force X
ticular behavior during which they are active. Fish need change in length) the muscle generates during one cycle of
both kinds of muscles if they are to perform both slow shortening and lengthening (Figure 10-38A). Net work is
swimming and fast escape responses optimally. graphically equivalent to the area contained within the
force-length loop (Figure 10-38B). A muscle generates pos-
Kinetics of activation and relaxation itive work only when it is shortening; thus positive work is
In considering the one-shot jump of frogs, our main con- equal to the area under the force-length curve during the
cern was to determine whether the muscle becomes maxi- shortening phase of a cycle. A muscle generates negative
mally activated during the early phase of shortening. The work when it is forcibly lengthened by an antagonist mus-
kinetics of muscle relaxation were essentially irrelevant. A cle (or a servomotor); thus negative work is equal to the
fundamentally different problem is faced by animals dur- area under the force-length curve during the lengthening
ing cyclical locomotion, such as swimming by fish. Swim- phase of the cycle. The net work-the difference between
ming will be most efficient if muscles do not have to work the positive and negative work done during one cycle-is
against one another. For example, when muscles on one the area between the positive and negative legs of the force-
Length
388 PHYSIOLOGICAL PROCESSES
..............................................................................
length curve for one cycle. For the muscle to generate Stimulus
net positive work, it must generate a greater force during
shortening than was required to stretch it to its initial
length. The net power generated in a cyclical contraction Length /\
is expressed by:
Force 1
(positive work-negative work)cyc,,X frequency of cycles
will reduce the amount of work the muscle can do. Once
again, there is a trade-off between two desirable features.
In this case, it is the ability of the muscle to do work ver- Muscles from different locations along the axis of the
sus its metabolic efficiency. Workloop experiments have body receive different stimulus patterns and change in
been performed on swimming fish to determine whether length by different amounts, which affects both the force
the swim muscles emphasize rapid relaxation, which is generated and the power produced. The stimulus duty cy-
metabolically costly, or lower work output, which is less cle (the percentage of one cycle during which the muscle is
metabolically costly. stimulated) is about 50% in the anterior part of the fish and
The basic experimental approach used in these studies falls to only about 25% in the posterior part of the fish. In
was similar to that described for the frog hip flexor muscle. addition, posterior muscles change in length much more
The electrical activity of muscles and changes in muscle than do anterior muscles during swimming. The combina-
length were determined in swimming fish. Then using the tion of big changes in length and a short duty cycle permits
type of setup illustrated in Figure 10-34 isolated muscles the posterior muscles to generate a great deal of mechani-
were stimulated identically with the electromyogram, and cal power. When isolated anterior muscles were exposed to
the length of the muscles was controlled to match the the same set of conditions (i.e., stimulation pattern and
changes measured during swimming. The force and power length changes),they generated the same amount of power
generated by the muscles under these conditions were as the posterior muscles; it is the pattern of muscle behav-
determined, and the work done by the muscles then was de- ior at the back of the fish that generates more power, not an
termined by plotting the workloop. These experiments re- intrinsic property of the muscle fibers themselves.
vealed that the posterior muscles do more net work than an- Examination of the workloops for red muscles during
terior muscles during slow steady swimming (Figure 10-39). swimming indicates that the swim muscles operate with a
MUSCLES AND ANIMAL MOVEMENT 389
......................................
slow activation rate and a slow relaxation rate. Stimulation traction, must enter the myoplasm rapidly and be removed
to the posterior muscles, which generate the most power, rapidly (Figure 10-40, steps 1and 4). Second, myosin cross-
begins during lengthening and ends just after the beginning bridges must attach to actin and generate force soon after
of shortening, as predicted for this kind of muscle earlier in the myoplasmic Ca2+concentration rises (steps 2 and 3)
this section. As a result, the muscle must be relaxing and then detach and stop generating force soon after the
through most of its power stroke, reducing the mechanical Ca2+concentration falls (steps 5 and 6). Myoplasmic free
power it generates, but presumably also decreasing the en- Ca2+in the red and white muscles of toadfish rises and falls
ergetic cost and perhaps thereby increasing the efficiency. with typical kinetics, but the Ca2+ transient in the sonic
muscles is the fastest ever measured for any fiber type
Adaptation for Speed: Sound Production (Figure 10-41A). Similarly, force measurements indicate
Some animals produce sound through mechanisms-for that the sonic muscles both contract and relax about 50
example, the movement of a column of air past a vibrating times faster than the red muscles (Figure 10-41B).
membrane or past the vocal cords-that are not directly The effect of the very fast Ca2+transient measured in
coupled to muscle contraction. In other animals, however; sonic muscles is most obvious during repeated stimulation.
sound is produced when muscles generate forces that di- When red muscle is stimulated at 3.5 stimuli per second
rectly cause structures, such as the swimbladder of a toad- (3.5 Hz), Ca2+lingers in the myoplasm so long that my-
fish or the rattle on the tail of a rattlesnake, to vibrate. In oplasmic free Ca2+cannot return to its resting value be-
these animals, the sound-producing (or sonic) muscles must tween stimuli. Indeed, the myoplasmic Ca2+concentration
undergo contraction-relaxation cycles at the frequency at remains above the threshold required for the generation of
which the sound is made, which is 10 to 100 times faster force for all of the time between stimuli, so a partially fused
than most locomotory muscles operate. tetanus is produced (Figure 10-42A). By contrast, the
In the last section, we saw that the swim muscles of fish swimbladder sonic muscle has such a fast Ca2+transient,
have relatively slow rates of relaxation, allowing them to that even at 67 Hz, the myoplasmic Ca2+concentration re-
avoid the high energetic cost of excessive calcium pumping. turns to its baseline value before every stimulus. Because
When these swim muscles are experimentally stimulated at the myoplasmic free Ca2+ concentration is below the
the high frequencies needed for sound production, they are threshold for the generation of force for much of the time
unable to relax between stimuli and therefore contract between stimuli, only the first two twitches in the series are
tetanically (see Figure 10-28).If the sound muscles became fused (Figure 10-42B). The production of an individual
tetanic in the same way, it would render the animal inca- twitch in response to each stimulus is required for generat-
pable of making any sound. Sonic muscles must then have ing the oscillation of the swimbladder that produces sound.
unique properties that allow them to operate at the high The ability of a muscle to relax rapidly requires
frequencies associated with sound production, which some- not only a brief Ca2+ transient in the myoplasm, but
times exceed 80 cycles per second (hertz, or Hz). also the rapid release of bound Ca2+ from troponin (see
Figure 10-40, step 5). Mathematical modeling indicates
Toadfish swimbladder that if the time constant for the release of Ca2+from tro-
The male toadfish, Opsanus tau, produces a "boatwhistle" ponin were the same in sonic fibers as it is in the fast white
mating call ten to twelve times per minute for many hours fibers of frog, twitches in sonic muscles could not be as fast
to attract females to its nest. This tone is generated by os- as those that have been observed. Comparisons of the time
cillatory contractions of the muscles encircling the fish's course of force generation and of myoplasmic Ca2+tran-
gas-filled swimbladder, which is described in detail in Chap- sients suggest that the release of Ca2+ from troponin in
ter 13. The steady swimming movements of toadfish occur sonic muscles of the toadfish swimbladder must occur three
at about 1-2 Hz, and their rapid escape response occurs at times faster than it does in frog white fibers.
5- 10 Hz. In order to produce sound, however, the muscles Finally, for force to drop quickly following the dissoci-
of the swimbladder must contract and relax at frequencies ation of Ca2+ from troponin, the myosin cross-bridges
' of several hundred hertz. To understand the differences must detach rapidly from actin filaments. The Huxley
among these various types of muscles that allow them to model discussed earlier in this chapter suggests that the
function with such different time courses, the properties of maximal velocity of shortening of a muscle, V,,,, must be
each of their three kinds of muscle-swim, escape, and proportional to the rate at which cross-bridges detach from
sonic-have been studied. The time course of many bio- actin. Indeed, the Vmaxof swimbladder sonic muscle (about
logical events is characterized by the half-width, which is 12 muscle lengths per second) is exceptionally fast, being
the width on the temporal axis of the event when the mea- five times higher than toadfish red muscle and two and a
sured variable is equal to half of its peak value. The half- half times higher than toadfish white muscle.
width of a single twitch is 500 ms in red (swim) muscle, Experiments with toadfish indicate that sonic fibers have
200 ms in white (escape)muscle, and only 10 ms in swim- a number of adaptations that permit them to operate at very
bladder (sonic) muscle. high frequencies. Ultrastructural and biochemical studies,
If a muscle is to activate and to relax rapidly, two con- for example, suggest that the short Ca2+ transient is
ditions must be met. First, Ca2+,the trigger for muscle con- achieved by an unusually high density of calcium channels
-
390 PHYSlOLOGlCAL PROCESSES
......................................................
Figure 10-40 In muscles that operate at
high frequencies, critical steps in the con-
traction-relaxation cycle must occur very
Transverse
rapidly. During activation, the stimulating sig-
tubule Sarcoplasmic
nal (dashed arrow) is conducted down the
clsterna reticulum
T tubule and communicated into the sar-
coplasmic reticulum, leading to opening of
calcium channels in the SR membrane and
e
w
"
u Ca2+
ATP ADP+ P,
Ca2+ outflow into the myoplasm (step I).
Binding of the free Ca2+to troponin (step 2)
relievesthe tropomyosin-mediated inhibition
of binding between actin and myosin.
Myosin cross-bridges then attach to the actin
filaments (step 3), and the thick and thin fila-
ments sl~depast one another. During relax-
ation, calcium pumps in the SR membrane
resequester Ca2+(step 4). The resulting drop
in myoplasmic Ca2+favors release of bound
Trooonin \ /
Thin filament
Thick filament
/
1 I I I
0.0 0.2 0.4
Time (s)
I I I I
0.0 0.2 0.4 0.6 0.8
Time (s)
Figure 10-41 Because sonic muscles of toadfish activate and relax much sured under the same conditions as in part A. Sonic muscles both con-
more rapidly than red (swim) and white (escape)muscles, they can oper- tract and relax much faster than do the red or white fibers. The average
ate at higher stimulation frequencies without becoming tetanic. (A) Time half-width of twitch tension ranges from 9.5 to 516 ms. Thus sonic muscle
course of myoplasmic Ca2+concentration in three types of muscle fibers operates more than 50-fold faster than red muscle. The Ca2+concentra-
isolated from toadfish following stimulation at 16°C. The average half- tion and tension records are normalized to their maximal values for all
width of the Ca2+transients ranges from 3.4 to 110 ms, as indicated on fiber types. [Adapted from Rome et al., 1996.1
the graph. @)Time course of twitch tension in the three fiber types mea-
MUSCLES A N D A N I M A L M O V E M E N T 391
...................................... ......................................
A Redfibers B Son~cfibers
I
0 1 2 3 4 5
Time (s) Time (ms)
Figure 10-42 Red muscle from a toadfish contracts tetanically in re- erated by a sonic fiber from a toadfish swimbladder stimulated at
sponse to relatively low-frequency stimulation,whereas sonic muscle pro- 67 Hz. Although the threshold Ca2+concentration for activation (dashed
duces individual twitches, even when stimulated at a much higher fre- line) is much higher in the swimbladder sonic fiber than in the red fiber,
quency. (A) Myoplasmic free Ca2+in and force generated by a red fiber the Ca2+transient is fast enough that the concentration falls below the
from a toadfish stimulated at 3.5 Hz. The threshold concentration of my- threshold value between each stimulus. Notice the difference in time
oplasmicfree Ca2+necessaryfor the generation of force is shown by the scale between parts A and B. [Adapted from Rome et al., 1996.1
dashed line on the Ca2+trace. (B) Myoplasmicfree Ca2+in and force gen-
in the SR membrane through which Ca2+is released; an un- suggests that these muscles might have many features in
usually high density of calcium pumps through which Ca2+ common with the toadfish swimbladder sonic muscles.
is resequestered; an increased concentration of certain cal- Indeed, it has been found that the fibers that shake
cium-binding proteins (e.g., troponin); and a fiber mor- the rattle (shaker fibers) have a very rapid calcium tran-
phology such that the distance between the SR membrane sient, with a half-width of 4-5 milliseconds at 16"C, only
and the myofilaments is particularly short, reducing the time 1-2 milliseconds slower than that of swimbladder sonic
required for diffusion. The rapid release of Ca2+from tro- fibers (Figure 10-43A).However, at 16°C the half-width of
ponin likely reflects decreased affinity of troponin for Ca2+. the shaker muscle twitch is considerably longer than that of
Finally, the rapid detachment of cross-bridges implies that swimbladder (Figure 10-43B). The twitch of the shaker
myosin in sonic fibers also has special molecular properties. muscle is slower probably because its cross-bridges detach
These adaptations allow swimbladder sonic muscles to more slowly; the V,, of the shaker muscle is about 7 mus-
perform mechanical work at high operating frequencies. To cle lengths per second, only about half the V,, of the
emit continuous sound, sonic muscle must generate work swimbladder muscle. In addition, Ca2+may detach from
to overcome frictional losses in the sound-producing sys- troponin more slowly, but this feature has yet to be deter-
tem and to produce sound energy. Workloop experiments, mined. The properties of rattlesnake shaker muscle suggest
similar to those described previously for the swim muscles that a rapid calcium transient alone is not sufficient for the
of fish, show that swimbladder fibers can perform work at production of very rapid contractions. The release of Ca2+
frequencies above 200 Hz at 25"C, the highest frequency from troponin and the detachment of cross-bridges from
for work production ever recorded in vertebrate muscle. By actin filaments must be unusually fast as well. For example,
comparison, the highest frequency known for vertebrate lo- at 16°C shaker fibers can be stimulated up to only about
comotory muscles is 25-30 Hz, measured in mouse and 20 Hz before summation of force begins, with tetanic fu-
lizard fast twitch muscles at 35°C. sion occurring at about 50 Hz. (Rattling occurs at 30 Hz at
16°C.)In contrast, sonic fibers produce individual twitches
Rattlesnakes at 67 Hz at 16°C.
Rattlesnakes in the genus Crotalus also use special noise- However, many snakes are active at temperatures
making muscles, but as a warning to members of other above 30°C, and they rattle at 90 Hz at a temperature of
species rather than to attract conspecifics for mating. Rat- 35°C; at this temperature, the calcium transient and the
tling is a loud and effective warning that renders these twitch speed are even faster in the shaker muscle than in the
snakes, like many venomous animals, very conspicuous. swimbladder sonic muscle at 16°C (see Figure 10-43).
Unlike the periodic toadfish boatwhistle, rattling can be Most likely, both the V,, of shaker fibers and the rate at
sustained continuously for up to 3 hours. However, the ra- which Ca2+is released from troponin are higher at 35°C
pidity with which the rattle-producing muscles contract than at 16°C. At 3S°C, shaker fibers can be stimulated at
392 P H Y S I O L O G I C A L PROCESSES
.............................................................................
A Shaker, 35°C B Sonic, 16°C
1 (1.4 ms)
(8.5 ms) ';
\:
0 10 20 0 10
Time (ms) Time (rns)
Figure 10-43 At 16"C, rattlesnake shaker muscles have a comparable more rapid. (B) Time course of twitch tension in toadfish sonic fibers
calcium translent t o toadfish sonic muscles, but their twitches last longer. and rattlesnake shaker fibers measured under the same conditions as in
(A) Myoplasmic free Ca2+following stimulation of toadfish sonic fibers part A. A t I U C , the half-width of twitch tension is nearly three times
and rattlesnake shaker fibers at the indicated temperatures. The half- longer in shaker fibers than in sonic fibers, but at 35OC, the shaker fibers
widths of the calcium transient, indicated in parentheses, are quite simi- contract and relax considerably more rapidly. The Ca2+concentration and
lar in sonic and shaker fibers at 16'C; however, at 35'C, a typical ambi- force records are normalized t o their maximal values. [Adapted from
ent temperature for snakes, the calcium transient in shaker fibers is much Rome, et al., 1996.1
100 Hz without complete tetanus, and they can perform and in many cases, the effort does not need to be sustained
work at 90 Hz. The similarities in the properties of toadfish for long periods. The situation is quite different for flight by
swimbladder sonic muscles and rattlesnake shaker muscles some insects, which depends on muscles that can operate at
suggest that in these species convergent evolution has ar- high frequency and produce considerable power. To power
rived at similar solutions to the problems that are posed by flight at wingbeat frequencies greater than 100 Hz, insects
high-frequency oscillatory contraction. have evolved special muscle fibers that can produce high
power at high frequencies. We discuss the properties of
Energetic and spatial constraints on muscle operation a t these high-power; high-frequency insect muscles in the next
high frequencies section.
The rapid calcium kinetics that are required in muscle fibers
that contract and relax rapidly could potentially demand
a relative increase in the surface area (and volume) of the
sarcoplasmic reticulum and in the number of mitochondria
within each fiber. Any such increase must reduce the space
that is available within each fiber for myofilaments, the
structures that generate force. In toadfish sonic fibers, for
example, the rate of Ca2+uptake by the sarcoplasmic retic-
ulum is about 50-fold greater than it is in red fibers, and
about 30% of the entire volume of a sonic fiber is occupied
by the sarcoplasmic reticulum. In addition, if a fast muscle
is to operate continuously, as the rattlesnake shaker muscle High-Power, High-Frequency Muscles:
does, it must use aerobic metabolism to generate enough Asynchronous Flight Muscles
ATP to fuel a high rate of calcium pumping; thus each fiber Most species in the Hymenoptera (bees and wasps),
must contain many mitochondria, displacing even more Diptera (flies),Coleoptera (beetles), and Hemiptera (true
myofilaments. bugs) contain flight muscles that are a notable exception to
In other words, there must be a balance between fea- the rule that no more than one contraction is evoked by a
tures that allow a muscle to operate very rapidly and the single depolarization of the surface membrane. This un-
amount of space left in each fiber for the contractile ma- usual type of striated skeletal muscle does not exhibit a one-
chinery-another example of a trade-off. If too much of to-one relation between the arrival of motor impulses and
the fiber volume is occupied by the components that sup- the timing of individual contractions. These flight muscles
port rapid calcium kinetics, the energetics of a fiber might are called fibrillar muscles, or more commonly asynchro-
be impressive, but there might be too few myofilaments to nous muscles, to distinguish them from muscles that con-
generate enough power to do the required work. tract in synchrony with each AP from a motor neuron. Al-
Vertebrate sonic muscles and some insect sonic muscles though the timing of contraction is unrelated to the timing
can operate at frequencies well above 100 Hz, but produc- of neuronal input to an asynchronous muscle, a constant
ing sounds does not require the production of high force, train of motor impulses and muscle depolarizations is re-
MUSCLES A N D A N I M A L M O V E M E N T 393
......................................
quired to maintain an asynchronous muscle in an active The mechanics of flight differ considerably in insects
condition. possessing synchronous flight muscles from those possess-
In some species of small insects, the wingbeat frequency ing asynchronous muscles. In those insects with synchro-
(and the frequency of wing-muscle contractions) far ex- nous flight muscles (e.g., damselfly), the wings are elevated
ceeds the maximal maintained discharge rates of which ax- and depressed by simple lever mechanics (Figure 10-4SA).
ons are capable. Wingbeat frequency has been found to Insects with asynchronous flight muscles have a more com-
vary inversely with wing size. A tiny midge, for example, plex musculoskeletal arrangement in which the thorax can
beats its wings at a frequency in excess of 1000 Hz, pro- exist in either of two stable configurations. In these insects,
ducing a high-pitched sound that can be perceived by the contractions of the antagonisticallyarranged flight muscles
human ear. change the shape of the thorax to generate only two wing
How can contractions take place in asynchronous mus- positions-up or down (Figure 10-45B).Contraction of the
cle fibers independent of the timing of the input stimula- elevator muscles pulls down the roof of the thorax, causing
tion? In some respects, asynchronous muscles are very sim- the wings to move up. As the roof of the thorax snaps down
ilar to the more-common synchronous muscles. For past an unstable "click" point (much as in a clicker toy), the
example, activation of asynchronous muscle requires a suf- depressor muscles are stretched and thereby activated, and
ficiently high concentration of free Ca2+in the myoplasm, the tension on the elevator muscles is suddenly released,
and there is some relation between neuronal input to the thereby relaxing them. Contraction of the depressor muscles
muscle and the myoplasmic Ca2+concentration. In fact, as shortens the thoracic exoskeleton front to back, so as to ex-
long as neuronal impulses continue to arrive at the neuro- pand the thorax dors~ventrall~, causing the wings to move
muscular junction, the myoplasmic Ca2+concentration is up. As the roof of the thorax moves back past the "click"
maintained at a steady activating level. However, even if position, the elevator muscles are stretched and thereby ac-
myoplasmic free Ca2+is elevated, the active state is not ini- tivated, and the tension on the depressor muscles is relieved,
tiated until the muscle is given a sudden stretch. The active thereby relaxing them. This alternation between elevator
state is terminated if tension on the muscle is released. This and depressor muscle activity can continue as long as con-
property has been demonstrated in glycerin-extracted asyn- ditions are appropriate for the thick and thin filaments of
chronous muscles. In the presence of a constant concentra- the fibers to slide past one another and generate force.
tion of Ca2+higher than lo-' M, an extracted asynchro- When neuronal input to asynchronous flight muscles
nous muscle actively develops tension if a stretch is applied, ceases, the muscle membrane repolarizes and the myoplas-
and it oscillates repeatedly between contraction and relax- mic Ca2+ concentration drops, with the result that the
ation if it is coupled to a mechanical system that has an ap- cross-bridges are unable to attach to the actin filaments. If
propriate resonant frequency (Figure 10-44). stretch is applied when myoplasmic Ca2+is too low, it no
I/
Time
394 P H Y S l O L O G l C A L PROCESSES
Depressors Elevators
contract
contract stretched
-.
-. -.
I I l l
Train of nerve impulses
396 PHYSIOLOGICAL PROCESSES
........................................
For example, if a small number of motor units in a signal in the muscle fiber (see Figure 10-46).The synaptic
muscle are maximally active, the muscle will contract with potentials along the distributed neuromuscular junctions
a small fraction of its total maximal tension. On the other are summed, and the shorter the interval between excita-
hand, if all the motor neurons innervating the muscle are tory synaptic potentials, the greater the depolarization of
recruited to fire at a high rate, all the motor units are the muscle membrane. Because the coupling between mem-
brought into a state of full tetanus, producing the maximal brane potential and tension is graded, each muscle fiber can
contraction of which the muscle is capable. In addition, produce a wide range of tension, instead of being limited to
many vertebrate muscles contain different types of fibers all-or-none twitches or tetanus, with no possibilities in be-
(see Figure 10-30), so the nervous system can modulate tween. For this reason, arthropod muscles function well
which fibers are active, as well as how many are active. over a large range of tensions with very few motor units,
Thus, the timing and force of a muscle's contraction is mod- with the variability in tension produced by single fibers re-
ulated by activity in the motor neurons innervating that placing the effect of recruitment in most vertebrate muscles.
muscle; by differentially activating motor neurons, the cen- In some arthropod muscles, one motor neuron may inner-
tral nervous system can determine the strength and dura- vate all-or at least most-of the fibers in the muscle.
tion of muscle contractions. The mechanical difference between twitch muscles and
The slow tonic muscle fibers of vertebrates (found tonic muscles is based on the manner in which myoplasmic
primarily in amphibians and lizards) are unusual in that Ca2+is controlled. In twitch muscles Ca2+is released from
they receive multiterminal innervation-that is, the motor the sarcoplasmic reticulum in an all-or-none manner in re-
neuron makes many synapses along the length of each fiber. sponse to all-or-none APs, whereas in graded tonic muscles
In these fibers, which lack all-or-none APs, the synaptic po- Ca2+ is released from the sarcoplasmic reticulum in a
tentials produced by the broadly distributed neuromuscu- graded fashion, because the electrical signals that are con-
lar junctions are sufficient to generate their graded con- ducted along the membrane are graded rather than all-or-
tractions (Figure 10-46B). The tension produced by these none (Figure 10-47).
muscles is strongly dependent on the frequency of motor In many invertebrates the flexibility of motor control
neuron activity, and these tonic muscle fibers generally are is further enhanced by multineuronal innervation of mus-
found where slow, sustained contractions are required. cle fibers. Each muscle fiber receives synapses from several
In mammals, only the extraocular muscles and intra- motor neurons, including one or two inhibitory neurons
fusal spindle fibers contain slow tonic fibers. As noted al- (Figure 10-48). The synaptic effects of inhibitory and ex-
ready, however, most skeletal muscles contain several types
of twitch fibers. Typically, all the fibers within a single mo-
tor unit are of the same type. In addition, the properties of Nerve impulse
the innervating motor neuron are often matched to the
properties of the muscle fibers. For instance, motor neurons
innervating slow oxidative (type I) muscle fibers carry im- Transmitter release
pulses at a lower frequency than do motor neurons inner-
vating fast glycolytic (type IIb) fibers.
Synaptic de~olarization
Motor Control in Arthropods
Arthropod nervous systems consist of a relatively small
number of neurons compared with vertebrates, so a small ~ r a d e fibers
d Twitch fibers
number of motor units must generate the full range of con-
tractions, from weak to strong, without relying on exten-
sive recruitment of new motor units. Moreover, many types Muscle action
of arthropod muscles never produce APs, or do so only un- I potential
der certain conditions. In these muscles, as in the tonic mus-
cle fibers of vertebrates, contraction is controlled by gaded
depolarization of the muscle fiber membrane rather than by
release
the frequency of muscle APs. The pattern of neuronal con-
trol that has evolved under these constraints is quite differ-
ent from the pattern of motor control in vertebrates.
Contractile
Each vertebrate twitch muscle fiber is innervated at activation
only one or two endplates, and postsynaptic potentials ini-
tiate APs that originate very near the endplate and are prop- Figure 10-47 Neuronal APs are linked to muscle contraction through ex-
citation-contraction coupling in the muscle fiber. In graded tonic muscle
agated along the muscle fiber. In contrast, crustacean skele-
fibers (left), the membrane potential varies based on summation and fa-
tal muscle fibers, like vertebrate tonic fibers, receive many cilitation of synaptic potentials at synapses distributed all along the mus-
synaptic terminals located along the entire length of the cle fiber In twitch muscle fibers (right), the muscle fiber membrane car-
muscle fiber, so no propagated AP is required to spread the ries APs, which give the contraction an all-or-none characteristic.
MUSCLES A N D A N I M A L M O V E M E N T 397
..............................................................................
Excitatorv axon most arthropod muscles of several types of muscle fibers
exhibiting different electrical, contractile, and morpho-
logic properties. At one end of the spectrum are fibers
with rapid all-or-none contractions, which resemble verte-
brate twitch fibers. When a series of intracellular current
pulses are delivered to such fibers, they produce a series
of subthreshold depolarizations until the firing level is
exceeded (Figure 10-49A). Once this occurs, the mem-
brane responds with an all-or-none AP, which then elicits
an all-or-none fast twitch. At the opposite end of the spec-
trum in crustacean muscle are fibers in which the electrical
responses show little sign of regenerative depolarization,
and the contractions are fully graded with the amount of
depolarization (Figure 10-49C). Between these two ex-
tremes is a continuum of intermediate muscle-fiber types
(Figure 10-49B).The differences in contractile behavior of
these fiber types are correlated with morphologic differ-
ences. The slowly contracting fibers have relatively fewer T
Figure 10-48 In contrast to vertebrate twitch fibers, each arthropod mus-
cle fiber receives synaptic input from several motor neurons. There are tubules and less sarcoplasmic reticulum than the rapidly
usually several excitor axons, and there may be one or more inhibitors as contracting fibers. As in vertebrates, the properties of mo-
well. Each motor neuron makes many synapses along the muscle fiber, tor neurons innervating each type of muscle fiber are at
producing a distributed neuromuscularjunction. One motor neuron of least somewhat matched to the properties of the fibers
each type is shown here for simplicity.
themselves.
Stimulating- r
current
e
- . tract very slowly. [Adapted from G. Hoyle,
1967, in Invertebrate Nervous Systems, C. A.
G. Wiersma, ed. O 1967 by University of
Chicago Press.]
Tension
CARDIAC MUSCLE liseconds long following the upstroke (see Figure 12-7).The
long duration of the cardiac-muscle AP and the associated
Cardiac muscle, the second type of striated muscle, shares long refractory period of several hundred milliseconds pre-
many characteristics with skeletal muscle but differs in sev- vents tetanic contraction and permits the muscle to relax, al-
eral important ways (Table 10-2). For example, vertebrate lowing the ventricle to fill with blood between APs. As a re-
skeletal muscle fibers are individually innervated by an ex- sult of regularly paced, prolonged APs, the heart contracts
citatory motor axon, whereas cardiac (ventricular) muscle and relaxes at a rate suitable for its function as a pump.
fibers in most but not all vertebrates are innervated only
diffusely by neurons of the sympathetic (excitatory) and
parasympathetic (inhibitory) divisions of the autonomic
system (see Chapter 11 for more discussion of the auto- . .
nomic nervous system). The cardiac innervation is modu-
latory only and does not produce discrete postsynaptic po-
tentials. Its actions are to increase or decrease the strength
of spontaneous myogenic contractions, which are induced As in skeletal twitch muscle, contraction of cardiac
by the electrical activity within the pacemaker region of the muscle is activated by an increase in the cytosolic Ca2+con-
heart (see Chapter 12). Another difference is that a cardiac centration. The rise in cytosolic Ca2+depends both on in-
muscle cell, or myocyte, contains one nucleus, whereas flux across the plasma membrane and release from the sar-
skeletal muscle cells are multinucleate. Cardiac muscle cells coplasmic reticulum. The cells of mammalian cardiac
are connected electrically such that an AP initiated in the muscle possess an elaborate sarcoplasmic reticulum and
pacemaker region spreads rapidly, from muscle cell to mus- system of T tubules (Figure 10-50).Membrane depolariza-
cle cell, through fast-conducting pathways to all muscle tion activates voltage-gated L-type calcium channels in the
cells within the heart, ensuring that the atria and ventricles T tubules resulting in an inward flow of Ca2+ from the
each contract as a unit. extracellular space. This small influx of Ca2+triggers the re-
Although the contractile mechanism of vertebrate ven- lease of a much larger pool of Ca2+from the sarcoplasmic
tricular muscle fundamentally resembles that of skeletal reticulum via calcium channels in the SR membrane, lead-
twitch muscle, their membrane APs differ. In contrast to the ing to contraction. Calcium is removed rapidly from the cy-
very short duration of the AP in skeletal muscle, the AP in tosol by calcium pumps in the SR membrane and by
cardiac muscle has a plateau phase that is hundreds of mil- Na+/Ca2+exchange proteins in the sarcolemma.
TABLE 10-2
Characteristics o f t h e major types o f muscle fibers in vertebrates
Mitochondrion
The relative importance of the SR and the plasma mem- larization producing greater tension (Figure 10-SlA).Re-
brane for Ca2+regulation varies among species. Cardiac duction of the extracellular Ca2+concentration leads to a
muscle of the frog has only a rudimentary reticulum and weaker contraction for a given depolarization, because less
tubular system. The myocytes of the frog heart are much Ca2+enters the cell (Figure 10-SIB).The intracellular Ca2+
smaller than adult mammalian cardiac muscle fibers, and concentration in cardiac muscle is determined not only by
their relatively large surface-to-volume ratio reduces the depolarization but also by a number of other factors in-
need for an elaborate intracellular reticulum for the storage, cluding the action of catecholamineson the heart. The cat-
release, and uptake of Ca2+.Instead, most of the Ca2+reg- echolamines epinephrine and norepinephrine that circulate
ulating contraction in amphibian heart cells enter through in the blood or are released from neuron terminals activate
the surface membrane as a result of the membrane's in- a- and P-adrenoreceptors on the surface of cardiac cells.
creased calcium permeability during depolarization. Adult Stimulation of a-adrenoreceptors activates the inositol
mammalian hearts, on the other hand, largely depend on phospholipid second-messengersystem (see Figures 9-14
calcium release from the sarcoplasmic reticulum. and 9-15), resulting in increased release of Ca2+from the
As in skeletal muscle, ryanodine receptors mediate ex- sarcoplasmic reticulum. In contrast, Padrenoreceptor stim-
citation-contraction coupling in mammalian cardiac mus- ulation activates the adenyl cyclase second-messenger sys-
cle. Low concentrations of ryanodine (in the nanomolar tem (see Figures 9-11and 9-12),resulting in increased cal-
range) lock the calcium channels in cardiac SR membrane cium flux across the sarcolemma. Thus stimulation of both
in the open state (see Figure 10-24).The Ca2+released from types of adrenoreceptors augments cardiac contraction.
the sarcoplasmicreticulum following ryanodine treatment The time course of cardiac contraction is determined by
at low concentrations is removed from myocytes by the duration of the increase in cytosolic Ca2+concentration
Na+/Ca2+exchange across the sarcolemma. The end result and the cross-bridgecycling rate, both of which may be tem-
is that SR calcium stores are reduced, the capacity to release perature dependent. Rapid cooling of the mammalian heart
Ca2+from the SR is diminished, and cardiac contractility from 30 to 10°C produces a prolonged contracture because
falls. Because the effects of ryanodine vary with the impor- the reduction in temperature slows the calcium pump in the
tance of the SR in regulating cardiac contraction, this drug SR membrane and Na+/Ca2+exchange across the sar-
has little effect on the contraction of the frog heart but a colemma, increasing the duration of the calcium pulse. Ani-
marked effect on the contraction of the adult rat heart. mals living at low temperatures can maintain high heart rates
The amount of tension that can be developed by a car- because they have enhanced calcium release and removal
diac muscle depends on the amount of Ca2+in the my- mechanisms compared with mammalian hearts at the same
oplasm. In the frog heart, when muscle cells are depolar- low temperature. Some animals, such as carp, that use their
ized, Ca2+ flows into the cell because of the increased muscles over a wide temperature range produce two differ-
calcium permeability of the depolarized membrane. Be- ent forms of myosin: a low-temperature (winter) form and
cause the influx of Ca2+is voltage dependent, tension de- a high-temperature (summer)form. These different forms of
velops as a function of depolarization, with greater depo- myosin allow a carp to maintain a reasonably stable time
400 PHYSIOLOGICAL PROCESSES
...............................................................................
A
A B Figure 10-51 The larger the de-
polarlzatlon and the hlgherthe ex-
320 -
/
tracellular Ca2+concentratlon, the
greater the tenslon developed In
Isolated frog ventricular muscle
0
fn
e
c - 240 - (A) Tens~on(upper traces) devel-
oped at three voltage steps (lower
16j/
c depolarlzatlon and extracellular
1.0 m~ Ca2+
c
I Z? Ca2+levels on tenslon developed
.-
*
0 3 1 I-"
The tens~onwas recorded at the
end of each voltage step and IS
.-WL 1 2 1 plotted against the membrane
m
-
o ! I ; potentla1 ~nrn~ll~volts [Morad and
a
80 0.2 r n Cazf
~ Orkand, 1971 ]
2- J
& I I I
- 40 0 + 40
Membrane potential (mV)
course of cardiac contraction throughout the year despite The innervation of smooth muscle differs significantly
wide fluctuations in temperature. from that of skeletal muscle, which has discrete and intimate
synaptic junctions between the motor terminal and the mus-
cle fiber. In vertebrate smooth muscle, the neurotransmitter
SMOOTH MUSCLE is released from many swellings, or varicosities, along the
Muscle fibers are called "smooth" if they lack the charac- length of autonomic axons within the smooth muscle tissue.
teristic striations produced by the organized groups of actin These do not form intimate junctions, but instead, the trans-
and myosin filaments that form sarcomeres. These least mitter released at a given varicosity diffuses over some dis-
specialized muscle fibers have myosin that is similar to the tance, encountering a number of the small, spindle-shaped
myosin found in contractile nonmuscle cells. The myofila- smooth muscle cells along the way. Receptor molecules on
ments of smooth muscle are gathered into bundles of thick the smooth muscle cells appear to be distributed diffusely
- -
and thin filaments in dense bodies, or they are connected to over the cell surface. Smooth muscle of vertebrates is gener-
attachment plaques located at the sarcolemma. The at- ally under autonomic and hormonal control and is generally
tachment plaques contain high concentrations of a-actinin, not "voluntary," as is control of skeletal muscle (one ex-
which is present in the Z disk of skeletal muscle, and the ception to this rule may be the urinary bladder). An inter-
protein vinculin, which is not present in the Z disk. In esting feature of single-unit smooth muscle is the sensitiv-
smooth muscle fibers, vinculin binds to the a-actinin and ity of the membrane to mechanical stimuli. Stretching the
anchors the actin filaments to the sarcolemma. muscle produces some depolarization, which in turn pro-
Vertebrate smooth muscles can be categorized into two duces some contraction. As a result, muscle tension is main-
categories: single-unit muscles and multi-unit muscles (see tained over a large range of muscle length. This response of
Table 10-2).In single-unit muscles, the individual muscle smooth muscle to stretch accounts, at least in part, for the
cells, which typically are small and spindle-shaped, are cou- autoregulation seen in arterioles; that is, contraction of
pled with one another through electrically conducting gap smooth muscle in the arteriolar wall in response to a rise in
junctions. If one or a few of the cells spontaneously depo- blood pressure maintains a reasonably constant blood flow
larize, the rest of the cells depolarize, too, because excita- in the peripheral tissues (see Chapter 12). Likewise, peri-
tion is passed through the gap junctions. As a result, a few staltic movements along the intestinal tract rely on stretch-
cells in single-unit smooth muscles can generate contraction induced contractions of single-unit smooth muscles.
that moves throughout the entire muscle in a wave. Neu- Smooth-muscle cells contract and relax far more slowly
rons synapse onto single-unit muscle cells and can modu- than striated muscle fibers and generally are capable of
late the rate and strength of contraction, but neuronal in- more sustained contractions. This is reflected in the dura-
put is not required for contraction. Single-unit smooth tion and amplitude of the cytosolic Ca2+pulse, which also
muscle forms the walls of vertebrate visceral organs (e.g., initiates contraction in smooth muscle, and the slower
alimentary canal, urinary bladder, ureters, and uterus). In process of excitation-contraction coupling, which is differ-
contrast, in multi-unit muscles each cell acts independently ent from that in striated muscle. The slow release and up-
and contracts only when it receives synaptic input from take of Ca2+ in smooth-muscle cells is associated with a
neurons. The muscles in the iris of the eye that regulate the poorly developed sarcoplasmic reticulum, which is com-
diameter of the pupil are multi-unit muscles. posed only of smooth flat vesicles close to the inner surface
of the celi membrane. A highly developed sarcoplasmic the greatest Ca2+influx and thus evoke the largest contrac-
reticulum like that of striated muscle fibers is unnecessary tions, because the tension generated is proportional to the
because the cells of smooth muscle are much smaller and intracellular level of Ca2+.
therefore have larger surface-to-volume ratios than striated Excitation-contraction coupling in smooth muscle oc-
fibers. No point in the cytoplasm is more than a few mi- curs by several different mechanisms. As discussed earlier,
crometers distant from the surface membrane. Thus, the regulation in striated muscle involves Ca2+binding to tro-
surface membrane of smooth-muscle cells can perform cal- ponin (see Figure 10-16), but smooth muscle lacks tro-
cium-regulating functions similar to those of SR mem- ponin. Rather, Ca2+ binds to calmodulin forming a
branes in striated muscle. Ca2+/calmodulincomplex (see Figure 9-17); this complex
In smooth-muscle cells, Ca2+is constantly pumped out- in turn binds to an elongated protein called caldesmon. In
ward across the surface membrane keeping internal Ca2+ the absence of Ca2+, caldesmon binds to actin thin fila-
levels very low. When the membrane is depolarized, it be- ments, restricting myosin-actin interactions and inhibiting
comes more permeable to Ca2+ions, permitting an influx muscle contraction. However, caldesmon that has been
of Ca2+,which activates contraction. Relaxation occurs phosphorylated by protein kinase C cannot bind to thin fil-
when the calcium permeability returns to its low resting aments and therefore does not inhibit myosin-actin inter-
level while the membrane pumps Ca2+out of the cell. Large actions. Thus phosphorylation of caldesmon or the binding
depolarizations of the membrane generate APs in which of Ca2+/calmodulinto caldesmon activates smooth-muscle
Ca2+carries the inward current. Action potentials produce contraction (Figure 10-52A).
cannot-
a curs. Binding of this complexto caldesmon
releases it from thin filaments, allowing the
bind to actin
PKC muscle to contract. Phosphorylation of
caldesmon by protein kinase C (PKC) also
prevents it from binding to thin filaments and
promotes contraction. (B) Binding of Ca2+to
the regulatory light chains of myosin allows
actin-myosin interactionsand promotes con-
B Ca2+binding to myosln light chams D Regulation of myosin light traction. (C) Phosphorylation of the regula-
chains by protein kinase C tory light chains by myosin LC kinase, which
is activated by Ca2+/calmodulin, also pro-
motes muscle contraction. (0).Phosphoryla-
tion of the regulatory light chains by protein
kinase C, at a site other than that acted upon
by myosin LC kinase, inhibits myosin-actin in-
teractions and causes smooth-muscle relax-
ation. [Adapted from Lodish et al., 1995.1
c
-
Phospholylation of myosin light chains
, +- -n i r
Phosphatase
Three other mechanisms for regulating smooth-muscle The head of a myosin cross-bridge hydrolyzes ATP, a
contraction involve the regulatory light chains of myosin. process that is significantly accelerated when the cross-
In smooth muscle and some invertebrate muscle, binding of bridge binds to actin. Hydrolysis of ATP, which requires
Ca2+directly to the regulatory light chains induces a con- Mg2+,produces a conformational change that permits the
formational change in the myosin head that allows it to cross-bridge to undergo a cycle of detachment, attachment,
bind to actin, so the muscle contracts (Figure 10-52B). and rotation on the actin filament. After a myosin head de-
Phosphorylation of the myosin light chains by myosin light- taches from actin, it can reattach farther along the actin fil-
chain (LC) kinase also causes contraction in vertebrate ament if conditions still favor attachment. The force-
smooth muscle (Figure 10-52C).Because myosin LC kinase generating rotation of the myosin head against the actin is
is activated by Ca2+/calmodulin,the rate of phosphoryla- believed to occur as multiple sites on the myosin head se-
tion is also calcium-dependent. Phosphorylation of another quentially bind to the actin filament. As successive sites
site on the regulatory light chain of myosin by protein bind and unbind, the rotation of the head against the actin
kinase C, however, induces a conformational change that filament stretches the attachment of the myosin head to the
prevents actin-myosin interactions, resulting in relaxation myosin rod, which then pulls on the thin filament, causing
(Figure 10-52D). Thus pho~phor~lation of caldesmon by it to slide past the myosin thick filament toward the center
protein kinase C and of the myosin regulatory light chains of the sarcomere. Because this process happens symmetri-
by myosin LC kinase results in contraction, whereas phos- cally at the two ends of the thick filaments, a sarcomere
phorylation of the regulatory light chains at another site by shortens symmetrically as the Z disks are pulled toward the
protein kinase C results in relaxation. The slowness in the center of the sarcomere. A number of muscle properties can
actions of protein kinases, along with the slow changes in now be understood in the light of this sliding-filament hy-
cytosolic Ca2+levels, contributes to the slow rate of con- pothesis of muscle contraction.
traction seen in many smooth muscles. At rest, myosin heads cannot bind to actin sites be-
cause the associated protein tropomyosin sterically hinders
binding. When a muscle fiber is depolarized, the AP prop-
agates into the T tubules, which, through a series of steps,
causes calcium channels in the SR membrane to open. As
a result, Ca2+is released from the sarcoplasmic reticulum
into the myoplasm and then binds to troponin, a multi-
subunit globular protein complex that is attached to both
actin and tropomyosin. When calcium binds to troponin,
Smooth-muscle contraction is modulated by a wide va- the protein undergoes a conformational change, which is
riety of stimuli, both neuronal and humoral, which can in- transmitted to tropomyosin, thereby revealing myosin-
hibit or activate contraction. All of these stimuli operate to binding sites on actin thin filaments. In this way, Ca2+reg-
influence cytosolic Ca2+levels andlor the level of protein ki- ulates contraction in vertebrate striated muscle. As the sur-
nase C, myosin LC kinase, and muscle phosphatases. As in face membrane repolarizes, the sarcoplasmic reticulum
cardiac muscle, cytosolic Ca2+levels in smooth muscle are begins taking up Ca2+again, removing it from troponin,
regulated by hormonal activation of the adenyl cyclase and terminating the active state of the muscle, and causing the
inositol phospholipid second-messenger systems. The di- muscle to relax as long as ATP is present. Although acti-
verse mechanisms that control smooth muscle result in the vation of smooth muscle also requires Ca2+,the mecha-
complex patterns of contraction observed in this type of nisms involved differ from that in striated muscle.
muscle. The mechanical and energetic properties of different
muscle fiber types exhibit remarkable diversity and adap-
tation to various activities. Muscular systems have evolved
so that muscles operate at the optimal myofilament overlap
SUMMARY to generate maximal force, and they shorten at the appro-
Muscles are classified into two major types: striated and priate velocity (VIV,,,) to generate maximal mechanical
smooth. Because striated muscle has been so intensively power with near-optimal efficiency. The kinetics of activa-
studied, its structure is perhaps better understood than that tion and relaxation tend to be relatively slow in locomotory
of any other tissue. It appears striated when viewed with a muscles, minimizing the high energetic cost of calcium
light microscope because it contains a regular array of par- pumping by the sarcoplasmic reticulum. In contrast, ver-
allel myofilaments, organized into banded sarcomeres. A tebrate sound-producing muscles must operate at very high
sarcomere contains an almost crystalline array of myosin frequencies, and thus have very rapid calcium pumping. Al-
and actin filaments. Myosin thick filaments interdigitate though sonic muscles are accordingly very expensive ener-
with and slide between the actin thin filaments. During getically, they only make up a small fraction of any animal's
muscle activity, the filaments slide past one another due to muscle mass. Finally, insect asynchronous flight muscles are
interactions between the actin filaments and cross-bridges able to generate high mechanical power at very high fre-
that project from the myosin filaments. quency while avoiding the high energetic cost of calcium
pumping, because they use stretch activation and shorten- Why do muscles become rigid several hours after an
ing deactivation to achieve high-frequency modulation animal dies?
of force. How is the force that causes the thick and thin fila-
The control of muscle tension by the nervous system ments to slide past one another produced by the
has evolved in several ways in different animal groups. myosin cross-bridges?
Most vertebrate striated muscle fibers respond to impulses When a muscle is shortening at V, ,what is the net
from a single motor neuron with all-or-none twitches, be- force generated by its cross-bridges? What is the
cause they contract when the fibers themselves generate all- power produced?
or-none APs. However, the muscle APs fuse into a steady Why does the velocity of shortening decrease as heav-
tetanic contraction if the stimulating impulses occur with ier loads are placed on a muscle?
high enough frequency. Many arthropod striated muscle Explain the steps by which Ca2+regulates contraction
fibers (as well as vertebrate tonic fibers) contract in a of striated muscle fibers.
graded, rather than in an all-or-none fashion, in response List the steps of muscle activation and of muscle
to graded, nonpropagated depolarizations at synapses that relaxation.
are distributed all along the muscle fiber. Most arthropod How can depolarization of the surface membrane of
muscle fibers receive inhibitory innervation, in addition to a striated muscle fiber cause Ca2+to be released from
synaptic input from several excitatory motor axons. the sarcoplasmic reticulum? What molecules are
Vertebrate cardiac muscle is organized at the myofib- involved?
rillar level much like striated skeletal muscle, but differs in What are the major processes in muscle function that
several ways. The fibers of cardiac muscle consist of many require ATP?
short individual cells electrically coupled to one another via What limits the tension that can be produced by a
gap junctions. In skeletal muscle, in contrast, embryonic myofibril?By a muscle fiber? By a muscle?
cells fuse into long multinucleated fibers, losing their in- What allows a muscle fiber to produce greater tension
tegrity as individual cells. The ionic mechanisms of cardiac during tetanic contraction than during a single
muscle are specialized for pacemaker activity in regions of twitch?
the atria and for prolonged APs in the ventricle. Define mechanical power. Define efficiency. Why are
Multi-unit smooth muscle is composed of indepen- mechanical power and efficiency equal to zero during
dently acting cells that require neuronal input to contract. isometric contractions and when a muscle shortens
Single-unit smooth muscle, which is more common in ver- at V, ?
tebrates, is composed of spindle-shaped cells that are elec- 17. During locomotion what is the disadvantage of using
trically coupled to one anothe-. Single-unit smooth muscle too slow a muscle fiber (VmaX~.
~-
is too low) to power a
is present in the walls of vertebrate visceral organs; it can movement requiring a given shortening velocity?
contract in response to stretch. All smooth muscle contains What is the disadvantage of using too fast a fiber
actin and myosin fibers, but they are not present in the or- (V,, is too high)?What is an optimal value of V,,,?
ganized manner characteristic of striated muscle. Smooth- 18. Describe the features of the fish muscular system that
muscle contraction is activated by Ca2+,which enters the enables it to produce both relatively slow movements
myoplasm primarily across the plasma membrane during with little backbone curvature as well as very fast
depolarization, rather than from the sarcoplasmic reticu- movements with large backbone curvature.
lum as in striated muscle. This source of Ca2+is feasible in 19. Why must the muscles that produce sound relax very
smooth muscle because the contractions are slow and be- rapidly? What are the adaptations that permit the
cause the small cells have a large surface-to-volume ratio muscle to relax quickly?
and small intracellular diffusion distances. 20. Why is a large energetic cost associated with rapidly-
relaxing muscles? How do insect asynchronous mus-
cles avoid some of this cost?
REVIEW QUESTIONS 21. What factors determine frequency of contraction in
1. Describe the organization and components of each of insect asynchronous muscle?
these structures: myofilaments, myofibrils, muscle 22. Discuss the major functional differences between
fibers, and muscle. skeletal, smooth, and cardiac muscle.
2. What kinds of evidence led A. F. Huxley and H. E.
Huxley to propose the sliding-filament hypothesis?
3. Draw a sarcomere and label its components.
SUGGESTED READINGS
4. Discuss the contributions of myosin, actin, troponin, Alexander, R. McN., and G. Goldspink, eds. 1977. Me-
and tropomyosin to contraction of striated muscle. chanics and Energetics of Animal Locomotion. Lon-
5. Predict the sarcomere length-tension graph of a don: Chapman and Hall.
muscle with the following filament dimensions: thick Alexander, R. McN. 1989. Dynamics of Dinosaurs and
filament, 1.6 pm; bare zone, 0.4 pm; thin filament, Other Extinct Giants. New York: Columbia University
1.1 pm; Z disk, 0.05 pm. Press.
404 P H Y S l O L O G l C A L PROCESSES
.........................................
Alexander, R. McN 1992. Exploring Biomechanics. New McMahon, T. A. 1984. Muscles, Reflexes, and Locomo-
York: Scientific American Library. tion. Princeton, N.J.: Princeton University Press.
Ebashi, S., K. Maruyama, and M. Endo, eds. 1980. Muscle Rome, L. C., R. P. Funke et al. 1988. Why animals have dif-
Contraction: Its Regulatory Mechanisms. New York: ferent muscle fibre types. Nature 355:824-827.
Springer-Verlag. Stein, R. B. 1980. Nerve and Muscle. New York: Plenum.
Huxley, H. E. 1969. The mechanism o f muscular contrac- Taylor, C. R., E. Weibel, and L. Bolis, eds. 1985. Design
tion. Science 164:1356- 1365. and Performance of Muscular Systems. Journal of Ex-
Josephson, R. E. 1993. Contraction dynamics and power perimental Biology, Vol. 115. Cambridge: The Com-
output of skeletal muscle. Annu. Rev. Phsyiol. pany of Biologists, Ltd.
55527-546. Woledge, R. C., N. A. Curtin, and E. Hornsher. 1985. En-
Lutz, G., and L. C. Rome. 1994. Built for jumping: the de- ergetic Aspects of Muscle Contraction.New York: Aca-
sign of frog muscular system. Science 263:370-372. demic Press.
uman beings have been studying-and making pre- innervated an effector cell (Figure 11-1A).For example, in
H d lctlons
. about-animal behavior for as long as the the pharynx of the nematode Caenorhabditis elegans, sin-
species Homo sapiens has existed. Although our current in- gle cells have been identified that probably serve both re-
terest in the origin and control of animal behavior stems at ceptor and motor output functions. In even simpler organ-
least in part from a search for models of our own behavior, isms, sensory and motor functions are carried out in one
our ancestors probably studied the topic to optimize hunt- cell (Spotlight
- -
11-1).
ing strategies and to minimize the chance that they them- In primitive animals, receptors and effectors may have
selves would become prey. In some respects, our curiosity been distributed throughout the organism, allowing each
about what animals do, and why they do it, may originate region of the body to respond to the environment relatively
from a pressing need to know what that lion will do next. independently, without necessarily activating other regions.
The complexity of this problem becomes evident when we This kind of distributed nervous system is found in modern
consider all of the processes that contribute to behavior. jellyfish and in the free-living coelenterateHydra. Howeveq
How is information that will result in behavior collected by as more-complex animals evolved, neurons became more
the various sensory organs? Where in the nervous system numerous, circuits became more complex, and the nervous
are decisions made, and where is coordinated action orga- system became compacted into a central nervous system.
nized?How is activity in the nervous system translated into Within the central nervous system, many neurons are lo-
effective behavior? Understanding the function of the ner- cated in close proximity, increasing the possibilities for in-
vous and endocrine systems is a prerequisite for answering terconnection. With many-even most-neurons being
these and related questions about animal behavior. located in the central nervous system, receptors and effec-
All behavioral acts are ultimately generated by activity tors in the periphery are connected to the central nervous
in motor neurons that causes muscles to contract. The be- system by long axons.
havior of an animal-the net movement caused by spatial A common simple reflex in modern animals is the
and temporal patterns of muscle contraction-is con- monosynaptic reflex arc (Figure 11-lB),in which a sensory
stantly modified in response to stimulation from the envi- neuron (the receptor) synapses in the central nervous sys-
ronment. Some of these responses are simple and pre- tem onto a motor neuron that innervates a muscle (the ef-
dictable reflexes. Other kinds of behavior depend strongly fector). This type of reflex comprises three elements: a sen-
h on information stored from past experience and are, there- sory neuron, a motor neuron, and muscle fibers. When the
fore, less predictable to an observer who has no access to sensory neuron becomes sufficiently activated, it excites the
the animal's memories. The "hardware" underlying all be- motor neuron and, hence, the effector muscle. All through
havior is composed of neuronal networks, or interconnect- animal phylogeny, these elementary components of the re-
ing circuits of neurons. Unlike electrical circuits, which are flex arc-sensory input pathways and motor neurons that
wired in a single way, neuronal networks are not "hard synapse onto muscles-are characterized by features that
wired." Instead, they exhibit plasticity, which is the ability have been conserved from the most primitive invertebrates
to be modified functionally, and even to some extent to the most complex vertebrates.
anatomically, in response to experience. Most reflex arcs contain more than one central synapse
The simplest neuronal network is a reflex arc, in which and are, therefore, polysynaptic pathways. Such a path-
sensory input is transmitted through some number of way includes at least one interneuron connecting the sen-
synapses to produce a motor output signal, which then sory and motor neurons (Figure 11-1C). In evolution, the
causes muscles to contract. It is possible that the primordial number of interneurons increased enormously as ani-
reflex arc may have consisted of a receptor cell that directly mals became more complex, a development that allowed
406 PHYSIOLOGICAL PROCESSES
.........................................
'?!A R e c e ~ t ocell
r
Stimulus
Lntra~nervous
~ o t d neuron
r system
Effector cell
(sensory)
neuron
Interneuron. -
\\
Figure 11-1 In simple reflex arcs, sensory receptors activate effector cells
through a small number of synapses. (A) In this primitive reflex, the re-
ceptor cell directly innervates and activates an effector cell. Some
chemoreceptor cells in the pharynx of the nematode C elegans proba-
bly serve this function. (B) A monosynaptic reflex arc consists of a recep-
tor neuron that synapses onto a motor neuron, which in turn activates
muscle fibers. This kind of reflex is called monosynaptic because it in-
cludes only a single synapse within the central nervous system. (C) This
more complicated reflex relies on several synapses in series. In parts B
and C, the gray circle encloses the part of the reflex arc that lies within the ~fferent Central nervous
central nervous system. (motor) neuron system
behavioral complexity to increase dramatically in higher mals would represent their surroundings quite differently.
animals. Accumulated evidence strongly suggests that if a Information about the distance between the bat and an ob-
species has a large number of neurons interposed between ject would be represented by the intensity of echoed sound
input and output neurons, this situation-by itself-con- or in its time of arrival; for the bird, information about dis-
fers great potential for learning. tance would be represented by relative focal planes, relative
Although many properties of motor neurons, which are position on the retina, and relative position of the two eyes.
the final common pathway for motor output, have been As a result, birds and bats use very different regions of the
conserved through evolutionary time, those of neuronal brain and mechanisms of information processing to inter-
systems that process sensory information have not been so pret sensory signals from their environment. The auditory
strongly conserved. Some elements of sensory transduction regions of the bat's brain are large and complex, whereas
are common to many senses (see Chapter 7), but the prop- the visual region is small; in contrast, the visual regions of
erties of central neurons that process sensory signals have the bird's brain are large and complex.
been exquisitely tuned by the circumstances of the species Despite such differences across sensory modalities,
and can differ dramatically, depending on which sensory there are general principles that apply to many sensory pro-
system has become most important in each species. For ex- cessing systems. For example, independent parameters of
ample, although both birds and bats fly-and thus en- the stimulus-such as the color, size, and direction of mo-
counter the same kind of environment-the way in which tion of a visual stimulus-are processed in separate, paral-
these animals encode information about their environment lel pathways. As information from each sensory modality
in sensory signals is quite different. As they fly in the dark, is transmitted through the brain, the properties of the stim-
bats glean information about their surroundings by emit- ulus that are represented in each brain region become in-
ting sounds and listening for echos reflected by surfaces. creasingly specific. In addition, most stimuli are organized
Most birds rely heavily on vision. If a bird and a bat were systematically within each region of the brain, generating a
to fly in the same area, the sensory systems of the two ani- map in which parts of the body, parts of the environment,
S P O T L I G H T 11-1 end of an ~ n d i v ~ d uorganlsm
al causes ~tt o swlm forward more
end causes ~tt o reverse direction
rapidly; a touch on ~ t anter~or
s
BEHAVIOR IN ANIMALS The dlrect~onin which
(part A of the accompanying ~IIustrat~on).
a Paramecium swims depends on the direction in which its cilia
THAT LACK A NERVOUS are beating; a reversal in the direction of progress is caused by a
reversal of the ciliary beat. What mechanism could account for
SYSTEM
such a reversal in response t o mechanical stimulation?
The behavior of multicellular animals depends on activity in the Experiments in the laboratory of Roger Eckert revealed that
nervous system, b u t protozoa produce several interesting be- the membrane of Paramecium includes stretch-activated ion
haviors, too, although these single-celled creatures lack any neu- channels that are differentially distributed: the channels in the
rons or muscles. Instead, events within the single cell serve the anterior membrane are Ca2+selective, whereas the channels in
same functions as d o sensory receptor cells, interneurons, mo- the posterior membrane are K+ selective (see illustration).
tor neurons, and muscles. Consideration of the mechanisms that Thus, in single-celled organisms, behavior is controlled by
allow these apparently simple organisms t o produce surprisingly changes in membrane potential caused when ion fluxes across
complex behaviors can b e a source of insight into the enor- membranes change, when ion channels in the membrane open.
mously conservative nature of evolution. Perhapsthe properties of neurons and muscles can b e regarded
The ciliate Paramecium produces an avoiding response if it as an abstraction of some of the capabilities of these multifunc-
bumps into an object or if it is touched. Atouch on the posterior tional "simple" organisms.
Time
Mechanical stimulation
@ opens Ca2+selective
channels
Ca2+enters,
depolarizing
the cell
Ca2+accumulates and
the ciliary beat reverses
-20
-40
Posterior stimulation
A Parameciumavoids objects with which it has collided by changing its ulation to the anterior end, the membrane depolarizes and produces a
direction and rate of swimming.(A)After it collides with an object, a Para- graded, but weakly regenerative, change in the membrane potential that
mecium backs up, changes direction, and swims away in a new direction is associated with reversal of the ciliary beat. (C) Mechanical stimulation
(left).The passage of time is indicated by sequential numbers. Mechani- of the posterior end opens Kt-selective channels, producing a graded
cal stimulation of the anterior end opens Ca2+-selectivechannels (right), hyperpolarizationof the membrane that accelerates the ciliary beat by an
increasing the internal free Ca2+concentration, which in turn reversesthe unknown mechanism. [Part A adapted from Grell, 1973; parts B and C
ciliary beat. (B)When the Ca2+channels are opened by mechanical stim- adapted from Eckert, 1972.1
-, Neural circuitry
I
Sensory Motor
Stimulus Behavior
Figure 11-2 Information processing within the central nervous system tion is then integrated with other sensory information, with genetic pro-
can be divided into functionally related compartments.Sensory input en- gramming, and with memories of past experience, eventually activating
ters from receptors and is filtered and processed. The sensory informa- neurons that in turn generate appropriate motor output.
or some other attribute of the stimulus (e.g., the frequency some basis for speculating about the routes by which more
of sounds) are arranged topographically in an orderly fash- complex nervous systems may have evolved from simpler
ion with respect to one another. The original view that these ones. Until recently, in fact, comparing the structure and
maps are static has been replaced by the understanding that function of nervous systems in animals from different phyla
to some extent their topography can be regulated dynami- was the only basis for inferring evolutionary relationships.
cally by use-dependent mechanisms. We consider these It is now also possible to compare DNA sequences that
principles in detail later in this chapter. have been conserved across species and to infer evolution-
Animals can change their behavior as a result of expe- ary relationships on the basis of how similar or different the
rience, which we call learning, and can store the informa- sequences are in modern species. This method of molecular
tion to be used in the future, which we call memory. Study phylogeny provides information that complements more
of the physiological (and potentially the anatomic) mech- conventional analyses based on the study of whole organ-
anisms that underlie learning and memory has focused on isms. Reconstruction of phylogenetic relationships based
changes at synapses. Recent insights into the cellular and on a comparison of DNA sequences requires that the fos-
molecular basis of learning and memory can now be incor- sils or living organisms being compared have in common
porated into our view of how behavior is generated. Both a particular molecule or molecules for which the nucleic
learning and the production of complex behavior patterns acid coding sequence can be determined.
must depend on the enormous amount of neuronal cir- At the cellular level, the nervous system seems to have
cuitry that lies between the relatively simple afferent sen- undergone surprisingly little modification in the course of
sory pathways and the efferent motor pathways. In higher evolution. The electrical and chemical properties of nerve
organisms, most neurons are central interneurons. In cells in vertebrates and invertebrates are remarkably simi-
Figure 11-2, this complex interface between sensory input lar (seeChapters 5 and 6). Many general principles of neu-
and motor output is represented as a single box separating ronal function were derived from studies of the relatively
the input and output neurons. Indeed, this part of the ner- simple nervous systems of invertebrates and lower verte-
vous system remains a "black box" that is understood in brates, which are more amenable to experiments than are
only the most fragmentary sense. The outwardly observ- the more complex nervous systems of higher vertebrates. In
able relations between stimulus and response are the do- particular, neurons in many invertebrate species are large,
main of behavioral psychology. In contrast, one goal of accessible, and readily recognizable from animal to animal,
neuroscientists is to understand how the connecting neu- making their activity relatively easy to record and to ana-
ronal circuitry works. One approach has been to study the lyze. Recently, it has even become possible to do biochem-
nervous systems of relatively simple animals to determine ical and molecular analyses of single neurons that have
what occurs in the time between the reception of a stimu- been identified in, and dissected from, invertebrate nervous
lus and the production of observable behavior. In this chap- systems.
ter, we examine some of the systems that are helping us to The anatomically simplest nervous systems consist of
understand what the nervous system does when it processes very fine axons that are distributed in a diffuse network
sensory information and generates patterned responses. (Figure 11-3).Such nerve nets are commonly found among
the coelenterates. These axons make synaptic contacts at
points of intersection, and a stimulus applied to one part of
EVOLUTION OF NERVOUS SYSTEMS
an organism produces a response that spreads in all direc-
The evolution of the nervous system cannot be recon- tions away from the point of stimulation. If the stimulus is
structed directly from the fossil record in the same way that repeated at brief intervals, conduction is facilitated and the
we can reconstruct the evolution of, say, the leg bones in signal spreads further. Very little is known about synaptic
vertebrates, because soft neuronal tissues leave little trace. mechanisms in diffuse nerve nets, because the axons are ex-
However, examination of the neuronal organization of in- tremely fine, making intracellular recording from them very
creasingly complex members of present-day phyla gives us difficult technically. However, even in the very simple nerve
served between homologous neurons in different individ-
uals of the same species.
Segmented invertebrates have nervous systems that
are somewhat decentralized, consisting of a ganglion
within each body segment. Each segmental ganglion usu-
ally serves the reflex functions of the segment in which it is
located, plus perhaps those of one or more adjacent body
segments. In addition, information is exchanged among
ganglia through trunks of axons, called connectives. The
result is a series of ganglia and connectives in the ventral
nerve cord that is characteristic of annelids and arthropods
(Figure 11-5; see also Figure 5-5).At the anterior end of the
ventral nerve cord in these phyla, one or more relatively
large clusters of neurons form a brain-or supergan-
glion-which receives sensory information from the ante-
rior end of the animal and which controls movements of
the head. In addition, neurons with their somata in the
brain often exert some control over other ganglia along the
ventral nerve cord and can contribute to coordinating
movements that require the body to work as a unit. This
coalescence of neurons at the anterior end of the animal,
where many sensory receptors are concentrated, is called
cephalization and is a common feature of central nervous
systems. (Not all brains are located at the anterior end of
an organism; for example, at the posterior end of its ventral
nerve cord, the leech has a tail brain that is even larger than
the head brain.)
The segmental ganglia of annelids and arthropods have
been used extensively for neuronal analysis because each
Figure 11-3 Axons in the nerve net ofthe jellyfish Aurelia can be seen on ganglion contains a relatively small number of neurons;
the subumbrellar surface when the organism is viewed in oblique I~ght. and, in many cases, several ganglia contain identical com-
The axons are arranged in a diffuse network that runs in all directions. plements of neurons. Thus, in principle, an analysis of the
They innervate muscles that cause the umbrella to contract. [Courtesy of
interactions of neurons in one segment can be generalized
A. Horridge.]
to many-even to all-other segments of the nerve cord.
For example, this approach has been useful in studying the
nervous system of the leech (see Figure 5-5), in which the
nets of coelenterates and ctenophores, there is evidence that ganglia are highly similar to one another. Despite the sim-
the neurons are organized into reflex arcs. plicity of structure in its nervous system, a leech can per-
A major early advance in the evolution of nervous sys- form such complex tasks as swimming to seek food or
tems was the organization of neurons into ganglia, or clus- crawling to escape danger, which makes it particularly well
ters of neuron cell bodies. Ganglia are found in the coelen- suited for studies of the cellular basis of behavior.
terates and are common throughout most of the animal The structure of the nervous system varies among the
kingdom. A ganglion (Figure 11-4A, C) consists of neu- other invertebrate phyla. In contrast with worms and
ronal somata that are organized around a mass of nerve arthropods, whose body plans are segmentally structured
' processes (axons and dendrites) called a neuropil. This and bilaterally symmetrical, an echinoderm typically has a
mode of organization permits extensive interconnectionsto nerve ring and radiating nerves arranged about its axis of
be made among the neurons, at the same time allowing symmetry. Perhaps as a result of their radial symmetry,
each neuron to produce a minimum number of collateral echinoderms lack any brainlike ganglion. Mollusks have a
processes-that is, side branches arising from the axon or nonsegmental nervous system consisting of several dissim-
axons. A neuropil may appear to be a randomly arranged ilar ganglia that are connected by long nerve trunks.
tangle of fine processes, but injection of markers or dyes Neurons in some molluskan species have contributed
into individual neurons (Figure 11-4B and D) reveals that greatly to our understanding of neuronal interactions. The
the major features of each particular type of neuron are or- ganglia of opisthobranch mollusks (e.g., the sea hare
ganized similarly from one individual animal to another, Aplysia) and nudibranchs (e.g., Tritonia) contain a number
even though fine details may differ. Moreover, physiologi- of neurons with extraordinarily large cell bodies; in
cal evidence shows that connections within the neuropil are Aplysia, the somata of some neurons are more than 1 mm
so orderly that identical synaptic interactions can be ob- in diameter. Individual giant neurons can be recognized
410 PHYSIOLOGICAL PROCESSES
...........................................
A
Anterior
connectives
Ax
Connectives \\I( w
'peripheral nerve / H
trunks 500 prn
Figure 11-4 Neuronal somata are organized into ganglia in many marker remains within the cell and diffuses into all of its branches. Note
species-including the h~ghervertebrates. This figure illustrates ganglia the numerous small branches on which synaptic contacts can be made
from members of two invertebrate phyla. In these ganglia, individual with similar branches of other cells. The two large axons enter the pe-
nerve cells can be readily identified from one specimen to another (A)A ripheral nerve trunks at left and the two smaller axons enter the connec-
segmental ganglion in the annelid leech Hirudo, showing the position of tives. (C) Schematicdiagram ofthe abdominal ganglion ofthe molluskan
individual nerve cell bodies. The paired connectives at the top and bot- sea slug Aplysia cal~fornica.(D) Morphology of a neuron (shown in color
tom of the diagram contain axons that connect neurons in the ganglion part C) injected with an intracellular marker. This neuron sends axonal
with neurons in the ganglia of adjacent body segments. The peripheral branches to all of the peripheral nerves shown at the bottom of part C.
nerve trunks emerging laterally carry motor and sensory axons to the vis- [Part A adapted from Yau, 1976; part B adapted from Muller, 1979; part
cera and muscles. (0)Mechanosensory neuron (shown in color in part A) C adapted from Kandel, 1976; part D adapted from Winlow and Kandel,
that has been labeled by injecting it with an intracellular marker. This 1976.1
visually from preparation to preparation, and they lend mata, enabling an experimenter to determine the properties
themselves well to long-term electrical recording, injection that characterize a particular type of cell and to estimate
of experimental agents, and isolation for microchemical the amount of variation that can occur from individual to
analysis. As in the nervous systems of annelids and arthro- individual.
pods, individual neurons in mollusks can be reliably iden- The most complex nervous system known among the
tified on the basis of their location and the size of their so- invertebrates belongs to the octopus. The brain alone is es-
In general, the nervous systems of invertebrate animals,
excluding the octopus, contain significantly fewer neurons
than do those of the vertebrates; for this reason, inverte-
brate nervous systems are often referred to as "simple."
However, superficial appearances can be deceiving, and the
functional sophistication of even relatively simple nervous
systems becomes apparent on closer examination. Later in
this chapter, we will consider examples of the behavior of
Optic nerve
invertebrates and the neuronal circuitry that underlies the
Supraesophageal ganglion behavior.
From observation of the variety of nervous systems in
Circumesophageal the animal kingdom, several principles of evolution have
connective
emerged:
Subesophageal ganglion
1. Nervous systems in all organisms are based on one
type of cell, the neuron. Although neurons are sculpted
into myriad shapes in the course of development, the
mechanisms of electrical signaling within the cell and
the nature of the chemical s~gnalsthat allow informa-
Thoracic ganglia
tion to be transmitted between cells are highly con-
served across phylogeny.
2. The organization of the nervous system evolved
through the elaboration of one fundamental pattern:
the reflex arc. Just as the neuron is the basic structural
unit of the nervous system, the reflex arc is its basic op-
erating unit. In its simplest form, a reflex produces a
stereotypical response to a particular sensory stimulus.
In the simplest reflex arc, a monosynaptic reflex, a sen-
sory neuron synapses directly onto a motor neuron,
Abdominal ganglia
which synapses onto muscle cells. Coordinated con-
traction of particular muscles produces behavior.
3. Through evolution, there has been a trend toward gath-
ering neurons into a central nervous system, connected
to peripheral sensory receptors and muscles by long ax-
ons. The organization of these networks favors one-
way conduction through neurons, from dendrites to the
axon and, thence, to the axon terminals, although the
biophysical properties of axons would enable them to
y
conduct either toward or away from the soma.
4. Complicated organisms have more neurons than do
simpler organisms, and their neurons are often con-
Figure 11-5 The ventral nerve cord of the crayfish Astacus illustratesthe
segmented arrangement of the nervous system in many invertebrates.
centrated in a brain, usually contained in the head.
There is one ganglion in each body segment. Connectives contain axons 5. As nervous systems became more complex through
that run between ganglia, and the nerve roots that link the ganglia with evolution, new structures were added on to older
structures in the periphery contain sensory and motor axons. Bilateral structures, rather than replacing them. As a result,
symmetry, another propertytypical of nervous systems in most phyla, also
structures that subserve new functions, which have
is illustrated in this diagram.
arisen more recently in evolution, are often seen to be
literally layered onto older, more primitive structures.
timated to contain lo8 neurons-compare this number 6. The size of particular regions in the brain of a species
with the lo5 neurons in the entire leech body. The neurons is usually related to the importance to that species of
in the octopus brain are arranged in a series of highly spe- sensory input into, or motor control out of, those re-
cialized lobes and tracts that evidently evolved from the gions. For example, in animals that depend primarily
more dispersed ganglia of the lower mollusks. If number of on vision for survival, the regions of the brain that
neurons in any way correlates with intelligence, the octo- process visual information are typically larger than all
pus should be fairly smart, and behavioral studies have other sensory areas. In nocturnal animals, other brain
shown that, by invertebrate standards, the octopus is in- regions-for example, those that process hearing or
deed quite intelligent. olfaction, which are independent of light-are largest.
412 P H Y S I O L O G I C A L PROCESSES
..............................................................................
ORGANIZATION OF THE VERTEBRATE The somatlc system is also referred to as the voluntary sys-
NERVOUS SYSTEM tern, because motor neurons of this dlvls~oncontrol skele-
tal muscles and produce voluntary movements. The auto-
The vertebrate nervous system is organized into identifiable nomic nervous system includes motor neurons that
structural and functional regions (Figure 11-6), although modulate the contraction of smooth and cardiac muscle
neurons in many regions may work together to process in- and the secretory activity of glands. The autonomic ner-
coming information and to generate appropriate behavior. vous system thus controls the "housekeeping" systems,
For example, the system is divided into the central nervous such as heartbeat, digestion, and temperature regulation.
system (CNS)and the peripheral nervous system (PNS).The The term autonomic literally means "self-managed," and it
central nervous system contains most of the neuronal cell was introduced when the relation between the autonomic
bodies. It includes the cell bodies of all interneurons and of nervous system and the more voluntary parts of the central
most neurons that innervate muscles and other effectors. The nervous system was poorly understood. We now know
peripheral nervous system includes nerves, which are bundles that the apparently automatic responses regulated through
of axons from sensory and motor neurons; ganglia that con- the autonomic nervous system are integrated and con-
tain the cell bodies of some autonomic neurons; and ganglia trolled within the central nervous system, just like the re-
that contain the cell bodies of most sensory neurons. (The sponses produced consciously through more voluntary
retina is contained entirely within the central nervous system, channels. The neurons of the autonomic system are them-
unlike other sensory neurons.) Nerves are afferent if they selves divided in two categories: sympathetic and para-
carry information toward the brain and efferent if they carry sympathetic, which differ from one another anatomically
information away from it. Many vertebrate nerves are mixed and functionally.
nerves, which include both afferent and efferent axons. A striking characteristic of vertebrate nervous systems
Efferent output from the central nervous system can be is their enormous redundancy; that is, there are a large
divided into two main systems: somatic and autonomic. number of individual neurons of every identifiable type.
Upper cervlcal
syrnpathetlc gangllon
M ~ d d l ecervlcal
Syrnpathetlc ganglla
Syrnpathetlc trunk
Splanchnic nerve 1
-1st lumbar nerve
Rarni cornmunicantes
d
1st sacral nerve
Figure 11-7 The structure of the vertebrate central nervous system is is not apparent in many structures of the brain, it is the basis of organi-
very elaborate in the head, but segmental organization is maintainedin zation within the spinal cord. It is also apparent in the organization of the
both the brain and the spinal cord. Diagrams of a ventral view of the frog cranial nerves. [AdaptedfromWiedersheim, 1907; Neal and Rand, 1936.1
(A) and the human (B) central nervous systems. Although segmentation
414 PHYSIOLOGICAL PROCESSES
.............................................................................
Figure 11-8 Each spinal segment is connected to the peripheryby a dor-
sal root, through which afferent signals enter, and a ventral root, through
which efferent signals leave. (A) Relation between the vertebral column,
the spinal cord, and the spinal nerves, wh~chemerge between the ver-
tebrae. Vertebrae are separated from one another by cushions of con-
nective tissue, the intervertebral discs. Regions of the sympathetic ner-
vous system that lie in the periphery are included in this diagram. Nervous
tissue is shown in color. (6)Diagram of a cross-sectionthrough a single
spinal segment. Neurons in a polysynaptic reflex arc that connect input
from skin receptor endings to output onto skeletal muscles are shown in
color. The pathway includes an interneuron in addition to the sensory re-
ceptor and the motor neuron. Note that the cell body of the sensory af-
ferent neuron resides in a dorsal root ganglion outside the spinal cord.
[Adapted from Montagna, 1959.1
spinal processes
B
Receptor
W h ~ t ematter
Effector
which receives information from, and sends information to, structures in the gray matter are not myelinated, so this cen-
the periphery through the dorsal and ventral spinal roots tral region lacks the shiny whiteness of the tracts. A fluid-
(Figure 11-8B).Seen in cross section, the spinal cord is bi- filled central lumen, the spinal canal, is continuous with the
laterally symmetrical. Ascending (sensory)and descending fluid-filled cavities within the brain, which are called the
(motor) axons are grouped around the outside surface of cerebral ventricles. The cerebrospinal fluid in these cavities
the cord and form well defined tracts. This outer region of has a composition similar to that of blood plasma.
the cord is called the white matter, named for the white ap- The regular organization of the spinal cord has facili-
pearance of the myelin that ensheathes many of the axons. tated its study by ne~roph~siologists. To a very large extent,
The more centrally located gray matter of the spinal cord afferent and efferent pathways are anatomically separated
contains cell bodies and dendrites of interneurons and mo- from one another (a pattern that was observed more than
tor neurons, as well as the axons and presynaptic terminals a century ago and that has been referred to as the "Bell-
of neurons that synapse onto these neurons. Most of the Magendie rule"). Afferent axons enter the central nervous
system through the dorsal spinal roots (see Figure 11-8B); A Optic lobe
(tectum) Cerebellum
efferent nerve fibers carry information out of the central
nervous system through the ventral roots. (There are minor
Cerebrum
\ \ A
exceptions to this rule. For example, fine unmyelinated sen-
sory afferents enter the spinal cord through at least some of
the ventral roots in the cat.) The cell bodies of spinal motor
neurons are located in the ventral gray matter, called the
ventral horn, on each side of the spinal cord, whereas the
cell bodies of interneurons that receive sensory input are lo-
cated in the dorsal gray matter, called the dorsal horn. Af-
ferent axons that synapse onto sensory interneurons within 6 Optic lobe
Epiphysis (tectum)
the cord arise from sensory neurons whose cell bodies lie in
the dorsal root ganglia outside the central nervous system. Cerebrum
(There is one dorsal root ganglion on either side of each
spinal segment.) The segregation of sensory and motor ax-
ons into the dorsal and ventral roots makes it possible to se-
lectively stimulate the sensory input into (or motor output
1
arising from) a single spinal segment. Alternatively, input to Olfactory lobe' -
a segment or output from it can be selectively eliminated by Optic tiact
transecting one of the spinal roots.
Many reflex connections are located in the spinal cord.
Later in this chapter, we consider two spinal reflexes, the
stretch reflex and the pain-withdrawal reflex. In addition,
neuronal connections that produce patterned movements
of locomotion-for example, walking, running, and hop-
ping-are located in the spinal cord. Although signals
from the brain can activate, suppress, or modulate behav-
ioral patterns that arise from activity in spinal circuits, it
appears that connections among spinal neurons are suffi-
cient by themselves to generate complex and coordinated
behaviors.
Cerebral hemisphere
Corpus callosurn 1
The brain
In all vertebrates, the brain contains many groups of neu-
rons with specialized functions, such as the reception and
processing of information from the eyes and the initiation
of movements that require coordination of the whole body.
In the higher vertebrates, the brain contains many more
neurons than does the spinal cord, and it exerts very strong
control over the rest of the nervous system.
- Adult structures
Cerebrum
Thalamus
Figure 11-10 The vertebrate brain begins as
three linearly arranged enlargements of the an-
terior neural tube, which then become elabo-
rated. Initially,the brain consists of the forebrain,
the midbrain, and the hindbrain. Later, thefore-
brain d~v~des into the telencephalon and the di-
encephalon; the hindbrain forms the meten-
Hypothalamus cephalon and the myelencephalon.At the right
is a partial listing of adult structures that arise
-
n +Tectum from the secondary vesicles. Sensory informa-
tion is typically received in structures derived
from the myelencephalon and metencephalon,
-
cerebellum is sent rostrally through structures derived from
the mesencephalon and diencephalon, and is
then sent on to the cerebral cortex, which de-
n Oblongata velopsfrom the telencephalon. Thus, although
morphogenetic changes in the brain obscure its
segmental origins, the pattern of information
processing echoes early segmentation.
matter is organized into layers that are parallel to the sur- The cortical regions that are purely sensory in function
face, each having a recognizable pattern of input and out- include the primary auditory, somatosensory, and visual
put. In addition, it is organized into functional regions cortical areas. (Primary projection cortex is the first loca-
(Figure 11-11).Some areas of the cerebral cortex contain tion in the cortex to which information from a particular
neurons that are purely sensory in function; that is, they re- sensory modality is transmitted.) The amount of brain
ceive sensory information, process it, and pass it on. Other space allotted to each of the sensory modalities is related to
regions are purely motor in function. In primitive mam- the principal habits of an animal species. For example, the
mals, such as rats, the sensory and motor areas account for primary somatosensory cortex-which receives informa-
almost all of the cortex. In contrast, the cerebral cortex of tion from neurons that sense stimuli such as touch, tem-
human beings and other higher mammals contains large re- perature, and pain-is relatively much larger in rats and in
gions that are neither clearly sensory nor clearly motor in tarsiers (a type of primitive primate) than it is in cham-
function. These regions, such as the frontal cortex, are re- panzees and human beings. The larger somatosensory cor-
ferred to as association cortex, and they are responsible for tex in rats and tarsiers correlates with their dependence on
such complex functions as intersensory associations, mem- collecting and discriminating tactile information. On
ory, and communication. the other hand, all primates-including human beings,
1C
cortex
c Chimpanzee D H u m a n being
a
Frontal cortex
p~tal
tex
C ] r ~ ~ 2 s e n s o r y [?Association
clat~on"funct~onsNot~cethat the relat~veamount of
assoclatlon cortex Increasesfrom rat to human be~ng
The frontal, temporal, and occ~p~tal reglons are la-
Primary beled on the three prlmate bra~nsIn all draw~ngs,the
Motor
auditory posterlor pole of the braln polnts to the rlght.
418 P H Y S I O L O G I C A L PROCESSES
..............................................................................
chimpanzees, and tarsiers-have much larger primary vi- tex (Figure 11-12A). More neurons are dedicated to pe-
sual cortices than do rodents such as rats. ripheral areas from which "important" information is re-
The size and location of cortical regions have been de- ceived-for example, the face and the hands-than to
termined in electrophysiological experiments in which ac- other parts of the body. Indeed, in human beings, approx-
tivity recorded from neurons within each site is correlated imately half of the somatosensory cortex receives input
with the application of specific stimuli. Perhaps the most from the face and hands, with the other half responsible for
dramatic of these experiments have been performed on hu- the remainder of the body surface. This remarkable dedi-
man beings. Many neurosurgical procedures are performed cation of resources to tactile information coming from the
while the patients are awake and relatively alert. (The cen- face and the hands correlates with the importance of these
tral nervous system itself has no pain receptors, so local areas in our daily lives. For example, the human central
anesthesia in the scalp and skull at the site of electrode en- nervous system requires detailed sensory information from
try allows the patient to remain comfortable and awake the hands to carry out the fine manipulations that we per-
during the surgery.) In these procedures, the stimulation of form with our hands when we do tasks and when we iden-
neurons within a particular region of sensory cortex evokes tify objects by touch. The map of sensory distribution, also
sensations in the patients, who are able to tell the surgeon called the sensory homunculus, differs in detail among
where in the periphery the sensation seems to arise and species, reflecting the different roles played by particular
which modality is being stimulated. These experiments, parts of the body. In raccoons, for example, more sensory
conducted in the course of therapeutic surgery, have sup- cortical area is dedicated to the forepawsthan to any other
ported the hypothesis that all sensation occurs in the cen- part of the body, presumably because these animals ma-
tral nervous system, primarily in the sensory areas of the nipulate their food and other objects extensively in their
cerebral cortex. daily lives.
The somatosensory cortex exemplifies the mapping A further elaboration of the general principle that space
of sensory input within the sensory cortex. This region of in the brain is organized according to the importance of in-
the cortex is organized so that different locations receive coming sensory information is particularly well illustrated
inputs from specific areas of the body. Sensations from by the somatosensory cortex of the star-nosed mole. In this
areas of the body that are adjacent to one another are animal, 11fleshy rays protrude from each side of the nose
mapped so that they are adjacent to one another in the cor- (Figure 11-13A, B), each covered with small swellings
A Sensory
Figure 11-12 Both sensory and motor cortical regions are organized into face and hands occupies about half of the entire human somatosensory
topographical maps of the body. (A) Map of a transverse section through cortex. (B)Map of a transverse section through the human motor cortex,
. the somatosensory cortex, depicting the location of neurons and their showing the distribution of cortical neurons that project to motor neurons
corresponding peripheral projections-that is, the places in the periph- ofthe brain andspinal cord, which in turn control adivityin specificskele-
ery where the stimuli are subjectively "felt." Information coming from the tal muscles.
BEHAVIOR: INITIATION, PATTERNS, A N D CONTROL 419
...............................................................................
Figure 11-13 Sensory receptors in the elaborate nose of the star-nosed mole
map onto specific regions of the sensory cortex. (A)The nose of a star-nosed
mole, seen from the front. (B) A higher-magnificationview of the fleshy rays sur-
rounding the right nostril. (C) A brain section, parallel to the surface of the so-
matosensory cortex, that has been processed for cytochrome oxidase. The pat-
tern of raylike stripes of darkly stained tissue matches the pattern of rays on the
nose. Medial, up; posterior, right. [Courtesyof K. Catania.]
of the skin under which lie arrays of receptors and nerve sual sensations. The relative size of these areas also corre-
endings. When it hunts for food, the animal waves these lates with their importance to the animal. For example, the
fleshy rays, bringing them into contact with the substrate. visual cortex in the tarsier occupies nearly a third of the
The projections from this remarkable organ were traced entire cortical surface, whereas it takes up only a tiny frac-
anatomically to the somatosensory cortex. Sensory input tion of the cortex in a rat (see Figure 11-11).Within these
from the rays crosses the midline of the mole; and, in the areas, the sensory world is arranged in a highly ordered
contralateral somatosensory cortex, there are eleven fashion. In the primary visual cortex, for example, the two-
wedge-shaped cellular regions, each of which receives input dimensional world that stimulates the retina is mapped di-
from one ray (Figure 11-13C).Apparently, each ray of the rectly onto the two-dimensional surface of the cortex; that
nose maps directly onto a stripe of neurons in the cortex, il- is to say, points lying near one another in the environment
lustrating the principle that, in some cases, the nervous sys- excite cells that lie near one another in the cortex. This
tem organizes sensory input by sending information from retznotopic mappzng was one of the first features of brain
specific sensory locations to morphologically distinct struc- organization to be discovered, and it has guided explo-
tures in the brain. Similar mapping has been reported for ration of other sensory systems. However, understanding
the projection from individual facial vibrissae (i.e., sensory how other sensory modalities are processed has presented
hairs) of mice and rats onto specific sets of neurons, called challenges. The visual scene and the area of the cortical sur-
barrel fields, in the somatosensory cortex. In species such as face can be directly represented in two-dimensional maps,
star-nosed moles or mammals that have prominent but mapping of the other senses requires more complex
"whiskers" on their faces, the somatosensory cortex spa- computations by the nervous system, as we will see.
tially sorts information from these readily identifiable sen- The motor cortex is located adjacent to the somato-
sory structures. Spatially sorting incoming sensory infor- sensory cortex, and it, too, exhibits topographical repre-
mation may be common among all species, even though it sentation of the periphery (seeFigure 11-12B).As in the so-
is less readily observable in species that lack such spectacu- matosensory cortex, the spatial distribution of neurons in
lar sensory organs. the motor cortex correlates with the location of the muscles
The auditory cortex of the temporal lobe and the visual that are controlled by the neurons. Neurons that are near
cortex of the occipital lobe (see Figure 11-11) are both neighbors in the motor cortex control muscles that are ,
purely sensory. Direct electrical stimulation of these areas neighbors in the body. In addition, many neurons are ded-
during neurosurgery evokes rudimentary auditory and vi- icated to controllingmuscles that make precise movements;
420 P H Y S I O L O G I C A L PROCESSES
......................................
fewer neurons control muscles that make only large, im- parasympathetic pathways dominate, lowering the heart
precise movements. For example, the muscles that move rate, diverting metabolic energy to housekeeping tasks such
the human fingers execute very detailed and finely tuned as digestion, and so forth. When an animal is threatened or
motions, and the amount of motor cortex dedicated to con- frightened, causing it to prepare for emergency activity, in-
trolling the fingers is very large. In contrast, the movements creased activity in sympathetic neurons inhibits the house-
of the toes are relatively simple and coarsely controlled, and keeping functions and enhances functions that can support
the amount of motor cortex dedicated to the toes is corre- intense physical activity. The heart rate becomes elevated,
spondingly quite small. Interestingly, the fine details of mo- levels of glucose in the blood rise, and blood flow to skele-
tor maps are somewhat plastic; they can change to some tal muscles increases. These two states-deep sleep and
extent, depending on use. alarm-are at opposite ends of a continuum. Most of the
The control of movement arises from activity in the time, the two systems are nearly in balance; as a result,
motor cortex, based on input from other areas of the cor- physiological variables such as heart rate are held at some
tex and the brain. The signal travels to the body by several intermediate value. A short list comparing some sympa-
pathways, including the corticospinal tract, which contains thetic and ~ a r a s ~ m ~ a t h eactions
t i c is given in Table 11-1.
the axons of neurons that have somata in the motor cortex The functional unit in both the sympathetic and
and that synapse in the spinal cord. (The convention of the parasympathetic divisions is the autonomic reflex arc.
naming tracts according to the location of the somata and Figure 11-14 shows a sympathetic reflex arc. The afferent
synapses is common in the vertebrate nervous system.) The side of an autonomic reflex arc is largely indistinguishable
number of synapses that separate neurons in the motor cor- from a somatic reflex arc, although the sensory neurons are
tex from the spinal motor neurons varies among species. likely to respond to different stimuli-for example, the
The pathway includes more synapses in lower vertebrates, concentration of glucose in the blood or the oxygen tension
and fewer in the higher vertebrates such as primates. In rab- in the tissues. The efferent side of an autonomic reflex arc
bits, for example, a signal from the motor cortex must be is quite different from a somatic reflex arc. In both divisions
transmitted across several synapses in the spinal cord be- of the autonomic nervous system, the motor output is car-
fore it reaches the motor neurons. In cats, few synapses sep- ried by two neurons in series (Figure 11-1SA).The soma of
arate the motor cortex from the spinal segment containing the first neuron, called the preganglionic neuron, is located
the target motor neurons; but, when the signal reaches the in the central nervous system, and the soma of the second
appropriate spinal segment, it must traverse several in- neuron, called the postganglionic neuron, lies in a sympa-
terneurons before it reaches the spinal motor neurons. In thetic chain ganglion (also called a paravertebral ganglion).
primates, some cortical motor neurons synapse directly Postganglionic neurons lie entirely outside of the central
onto spinal motor neurons, an arrangement that is thought
to confer special motor capacities on primates. However,
neurons that directly connect the motor cortex with spinal
motor neurons constitute only about 3% of the motor con-
trol neurons. Thus even in human beings, most motor con-
trol is achieved through less direct pathways.
Neurons controlling motor activity maintain a steady
background of low-level synaptic input to the motor neu-
rons. An increase in their activity synaptically activates mo-
tor neurons, which can produce forceful movements of the
limbs. This behavior occurs naturally when a subject con-
sciously generates a strong contraction of s muscle, which
is why the system is called the "voluntary" system. Move-
Preganglionic
neuron Postganglionic fi
ments can also be evoked experimentally in anesthetized
animals when clusters of neurons in the motor cortex are
directly stimulated with weak electrical current.
Eye
Radial muscles of iris Pupillary dilation
Iris sphincter muscles Pupillary constriction
Ciliary muscle (controlling Relaxation (focuses on distant objects) Constriction (focuseson close objects)
thickness of lens)
Lacrimal (tear) glands Stimulates production of tears
Salivary glands Stimulates production of a small amount Stimulates production of a large amount
of viscous saliva ("dry mouth") of dilute saliva
Arterioles Vasoconstriction, especially in vessels Little or no effect
supplying skin
Heart
Pacemaker cells Increases rate of heartbeat Decreases rate of heartbeat
Ventricular contractile fibers lncreases force of contraction Little or no effect
Lungs
Bronchioles Dilates bronchioles Constricts bronchioles
Mucous glands No effect Stimulates secretion of mucus
Gastrointestinal tract
Sphincter muscles Contract~on Relaxat~on
Motlllty and tone of smooth muscles lnhlblts St~mulates
Exocr~nesecretion of glands lnh~b~ts Stimulates
Gallbladder lnhrbrts contractron Stimulates contraction
Liver Increases glycogenolyslsand, therefore, No effect
blood sugar
Urinary bladder No effect Contraction of muscles
Adrenal medulla Stimulates secretion No effect
nervous system, and they synapse onto the target cell of the rons is acetylcholine; the neurotransmitter of sympathetic
autonomic reflex. postganglionic neurons is norepinephrine. (Sympathetic
The location and properties of these postganglionic postganglionic neurons also release a little epinephrine.)
neurons depend on the autonomic branch to which they Postganglionic neurons of the parasympathetic and
belong (see Figure 11-15). In the sympathetic branch, pre- sympathetic branches typically innervate the same target
ganglionic neurons synapse onto postganglionic neurons in organs (Figure 11-1SB) and typically exert opposite effects
the sympathetic chain ganglia, which contain the cell bod- on their shared targets. For example, pacemaker activity of
ies of the postganglionic neurons. The axons of the post- the heart is slowed down by acetylcholine released by
ganglionic neurons then extend to the target organs, which parasympathetic postganglionic neurons, whereas it is ac-
may lie quite far away from the ganglion. (An exception to celerated by norepinephrine released by sympathetic post-
this pattern is the celiac ganglion, which contains the so- ganglionic neurons. These actions of the parasympathetic
mata of sympathetic postganglionic neurons innervating and sympathetic systems are reversed in the digestive tract,
the stomach, liver, spleen, pancreas, kidney, and adrenal where acetylcholine from parasympathetic neurons stimu-
gland. It is located in the abdominal cavity.) lates intestinal motility and digestive secretion, whereas
Preganglionic neurons of the parasympathetic nervous norepinephrine from sympathetic neurons inhibits these
system, on the other hand, synapse onto postganglionic functions.
neurons in ganglia that lie near-or even in-the walls of Within the autonomic nervous system, there are two
the target organs. Hence, in the parasympathetic branch, kinds of receptors for each of the neurotransmitters acetyl-
axons of preganglionic neurons may be very long, and ax- choline and norepinephrine. For each transmitter, the two
ons of postganglionic neurons are typically short. Pregan- types of receptors can be distinguished by drugs that act as
glionic neurons of the sympathetic nervous system are lo- agonists (i.e., mimic the natural transmitter) or as blockers.
cated in the cervical, thoracic, and lumbar regions of the The two kinds of cholinergic receptors are called nicotinic
spinal cord. The cell bodies of parasympathetic pregan- and muscarinic. Nicotinic and muscarinic receptors were
glionic neurons lie in the head and in the sacral spinal cord. first distinguished from one another on the basis of physio-
The two divisions of the autonomic nervous system logical reactions to two substances-nicotine and mus-
also differ pharmacologically. All preganglionic neurons are carine-that are never produced naturally in the body.
cholinergic; that is, their neurotransmitter is acetylcholine. Nicotine, a plant alkaloid, acts as an agonist at some
However, the neurotransmitter of a postganglionic neuron cholinergic synapses, including the synapses between pre-
depends on the division to which the neuron belongs. The and postganglionic neurons in both divisions of the auto-
neurotransmitter of parasympathetic postganglionic neu- nomic nervous system (see Spotlight 6-3). Muscarine,
422 PHYSIOLOGICAL PROCESSES
...............................................................................
Sympathetic Parasympathetic
Reproductive
system
Figure 11-15 The two divisions of the autonomic nervous system share breviation:ACh, acetylcholine. (B) Locations and targets of pre- and post-
many targets. (A) Both the sympathetic and the parasympathetic divisions ganglionic neurons in the sympathetic (left) and parasympathetic (right)
innervate their targets through a two-neuron chain. The soma of each branches of the autonomic nervous system. Preganglionic neurons are
preganglionic neuron lies within the central nervous system. The pre- shown in color; postganglionic neurons are shown as black lines. This di-
ganglionic neurons synapse in peripheral ganglia onto postganglionic agram illustrates the human autonomic nervous system, but the system
neurons that contact the target organs. The two divisions differ pharma- is similar in most vertebrates. Abbreviations: Cl, first cervical segment of
cologically.The preganglionic neurons of both divisions are cholinergic. the spinal cord; TI, first thoracic segment; L1, first lumbar segment; S1,
The postganglionic neurons of the parasympathet~cdivision also are first sacral segment; CX, coxal segment. In the parasympatheticnervous
cholinergic, but the postganglionic neurons of the sympathetic system system, several pathways run in cranial nerves, which are indicated by ro-
are adrenergic, primarily using norepinephrine as their transmitter. Ab- man numerals.
which is extracted from particular toxic mushrooms, acts some complex behaviors and then turn to some properties
as an agonist at other cholinergic synapses, including of networks known to underlie behavior.
synapses made by parasympathetic postganglionic neurons
onto their target cells. Curare (D-tubocurarine)blocks the Basic Behavioral Concepts
action of acetylcholine on nicotinic receptors (including A basic problem facing any animal is what to do (and not
those at the endplate of skeletal muscle), whereas atropine to do) in any given situation. Understanding the mecha-
blocks muscarinic receptors. Nicotinic and muscarinic re- nisms that allow animals to make these choices is a major
ceptors have completely different molecular structures and challenge. Behavioral scientists have taken two comple-
response mechanisms. Nicotinic receptors consist of pro- mentary approaches to meeting this challenge. The neu-
tein complexes that bind the neurotransmitter and contain roethological approach has been to bring the animal into
ion-selective channels within their structure. Muscarinic re- the laboratory and to observe behavior in this drastically
ceptors are molecules with seven transmembrane domains, simplified set of well defined circumstances. In the labora-
and they affect ion channels through the mediation of in- tory, there are no predators; the number and sex of con-
tracellular second messengers (see Chapter 6). specifics are controlled by the experimenter; there are un-
The adrenergic postganglionic neurons of the sym- usual lights, smells, and sounds; and the animal is often
pathetic division innervate two kinds of norepinephrine limited to a relatively small and confined space. In some
receptors, designated a and P receptors. Like the acetyl- cases, the nervous system of the animal is exposed surgi-
choline receptors, the adrenergic receptors are distin- cally to allow the experimenter to record from neurons
guished from one another by their pharmacology. Adren- while behavior is going on. Studies under these reduced
ergic a receptors are more sensitive to norepinephrine than conditions are useful, even necessary, for obtaining answers
to the drug isoproterenol, and they are selectively blocked to circumscribed, well defined questions about behavior,
by phenoxybenzamine. Adrenergic P receptors are more but the results from such experiments can be difficult to
sensitive to isoproterenol than to epinephrine and are se- translate into an understanding of how animals deal with
lectively blocked by propranolol. The two types of adren- challenges in their everyday lives. The ethologicalapproach
ergic receptors activate separate, but parallel, intracellular is to go into the field and observe the animal as it goes
regulatory pathways. about its business in its normal environment. Observing an-
Although all vertebrates have autonomic nervous sys- imals in their normal state is an ancient practice, and our
tems, the two divisions are not well defined in all groups. ancestors no doubt made such observations with dinner in
For example, in teleost fishes, sympathetic and parasym- mind. However, for the physiologist, these natural condi-
pathetic divisions are distinguishable; but, in cyclostome tions raise severe problems that differ from those created by
fishes, the autonomic nervous system is not partitioned laboratory conditions. While observing an animal in its
into two divisions. The autonomic nervous system of am- natural setting, it is difficult to determine the relative im-
phibians appears to be essentially identical with that of portance of all the different things that the animal does, and
mammals, but reptiles may not have such a clearly defined it is essentially impossible to record activity from the ani-
distinction between sympathetic and parasympathetic di- mal's nervous system. However, progress has been made in
visions. Comparative studies of the autonomic nervous sys- recognizing repeatable patterns in natural behaviors.
tem have been relatively rare, and further research may re- On the basis of observations made in the field and of
veal some phylogenetic surprises. many ingenious experiments, many workers have compiled
detailed inventories of what animals do and how and
where individual animals spend their time. These reports
ANIMAL BEHAVIOR offer a wealth of information but few organizing principles.
To understand how behavior can be initiated and con- From considering such observations, ethologists Konrad
trolled by the brain and other parts of the central nervous Lorenz and Niko Tinbergen developed an interpretive
system, it is necessary to know the properties of neurons framework that ultimately crystallized these data into a
and the ways in which information is transmitted from one useful system. They recognized that the behavioral reper-
neuron to the next. We have already considered how indi- toire of an animal seemed to be constructed from elemen-
vidual neurons are linked into circuits. The simplest neu- tal motor and sensory "units." They called the unit motor
ronal circuits are reflex arcs that depend on a small number patterns fixed action patterns (also called modal action pat-
of neurons, but the circuits that produce behavior can be terns), and the corresponding unit sensory elements they
immensely complex. .To understand these more compli- called key stimuli (also called sign stimuli). They were
cated circuits, we must consider several questions. First, awarded a Nobel Prize in 1973 in recognition of the im-
what exactly about behavior do we want to explain? Sec- portance of their observations.
ond, can we construct this explanation from known circuits Fixed action patterns have six properties:
and their interactions? Third, do general principles emerge
or is every behavior a "special case"? To suggest how 1. They are relatively complex motor acts, each consist-
clearly and completely these questions can currently be an- ing of a specific temporal sequence of components.
swered, in the remainder of this chapter we first consider They are not simple reflexes.
2. They are typically elicited by specific key stimuli rather these patterns has been used to deduce taxonomic
than by general stimuli. relationships.
3. Fixed action patterns are normally elicited by an en- 6. Fixed action patterns are typically performed in a rec-
vironmental stimulus; but, if an experimenter removes ognizable form even by animals that have had no prior
the stimulus after the animal's movements have begun, experience with the key stimuli. That is, these patterns
the behavior will usually continue to completion. This are inherited genetically, although it is clear that in
all-or-none property of fixed action patterns distin- many species the patterns can change somewhat with
guishes them from reflexes, which typically require experience. This property of heritability sparked a
continued stimulation to maintain movement. It is as long-lived debate about the dichotomy between inher-
though the key stimulus is required to turn the pattern ited and learned behaviors (i.e., "nature versus nur-
on; but, once begun, it plays out independently of fur- ture") that still flares up periodically like an incom-
ther stimulation. pletely extinguished fire.
4. The stimulus threshold for fixed action patterns varies
with the state of the animal, and the variation can be The stereotyped nature of muscle activity during fixed
quite large. For example, immediately after an animal action patterns has made them extremely useful to physi-
has copulated, it cannot be induced to copulate again ologists interested in the cellular basis of motor activity dur-
without intense stimulation. The threshold for elicit- ing behavior. The ability to reliably elicit precisely the same
ing copulation then drops as time passes. muscle contractions over and over again by applying the
5. When they are presented with the appropriate stimu- appropriate key stimulus has provided a window into the
lus, all members of a species (perhaps that are the same working of the nervous system, particularly in simple ani-
age or the same sex or both) will perform a given fixed mals. Some typical fixed action patterns are illustrated in
action pattern nearly identically. Some fixed action Figure 11-16.
patterns are common to many species of a genus. The Key stimuli, which specifically stimulate an animal to
properties of fixed action patterns are so reliable perform a fixed action pattern, have also been called re-
within some taxa that comparison of variations within leasers, because they appear to "release" a prepatterned be-
Figure 11-16 Fixed action patterns can be observed in most species. carried out by a domestic dog. (B) The courtship behavior of a male fid-
These behaviors appear to be genetically programmed, rather than dler crab. The crab attempts to attract a female by waving its large claw
learned. (A) Mating behavior in (1) octopus and (2) Helixpomatia. (3) Male (1)forward, then (2) laterally, then (3) upward, and finally (4)forward again.
Sepia officinalis in sexual display. (4) European wildcat striking with its [Part A, 2, 3, 5, and 6, adapted from Tinbergen, 1951, 1 adapted from
paw. (5) A digger wasp capturing its prey. (6) Male three-spined stickle- Buddenbrock, 1956,4 adapted from Lindemann, 1955, 7 adapted from
back quivering to stimulate a female to spawn. (7) The "mouse jump" Lorenz, 1954; part B adapted from Lorenz and Tinbergen, 1938.1
BEHAVIOR: I N I T I A T I O N , PATTERNS, A N D CONTROL 425
...............................................................................
havioral response within an animal. To sort out the features spring, or aggressive encounters, animals use complex sets
of a stimulus that are essential to its effectiveness, etholo- of movements to communicate their behavioral intent. In
gists have made models that are shaped and colored to courtship, for example, a movement by one animal sends a
mimic the suspected signal. The features are varied among signal to its partner. The partner may, in turn, move in a
the various models to determine which of them is most ef- way that sends a signal back.
fective at eliciting the particular fixed action pattern. A clas- Genetic components of fixed action patterns and key
sic example of a fixed action pattern that has been studied stimuli have been used to probe how species-specific be-
with the use of models is the aggressive response of a court- havior is organized. An example of this approach has been
ing male three-spined stickleback fish. When a courting the study of the wing movements that produce the chirps
male sees another male stickleback, it displays aggressively and trills in the courting song of a male cricket. Patterns of
and may even attack. Presenting courting male fish with cricket songs are species-specific and are largely indepen-
models that differed in shape and in color revealed that the dent of environmental factors other than temperature.
key stimulus for releasing this behavior is the red underside Moreover, the sound pattern produced by a species is di-
characteristic of male sticklebacks (Figure 11-17A). The rectly related to the pattern of action potentials (APs) in
shape of the model male fish was relatively unimportant, motor neurons that control the sound-producing muscles.
and the red underside was ineffective as a sign stimulus One set of studies examined sound production by males of
if the model was in any orientation other than horizontal two related species. One species produces a short trill (con-
(Figure 11-17B).Thus, it is not simply the color red but the sisting of about two sound pulses), whereas the other
red belly in the proper visual context that acts to release the species produces a long trill (consisting of about ten sound
aggressive behavior of the male fish. The discovery of these pulses). The trills of F, hybrids, the progeny produced by
key stimuli suggested further experimental work, just as the crossing individuals of the two species, consisted of an in-
discovery of fixed action patterns had. The highly specific termediate number of pulses (about four). Backcrosses to
nature of the releasing stimuli suggested that there might be produce various genetic combinations demonstrated that
adaptations within the stickleback's visual pathways that the neuronal network producing the song pattern is under
were specially tuned to important features, acting perhaps rigid genetic control, sufficiently precise to specify differ-
as sensory filters to admit only the appropriate parts of the ences as small as the exact number of APs going to the
stimulus. Indeed, the theme of specialized detectors has chirp-producing muscles.
proved to be an important and powerful organizing con- Similar experiments on vertebrate species have indi-
cept in sensory physiology. cated that, in these higher organisms, too, some aspects
Fixed action patterns and key stimuli do not occur in of behavior are genetically controlled and that hybrid in-
isolation but are instead woven seamlessly into behavioral dividuals perform an intermediate form of the behavior.
interchanges between animals. In courtship, caring for off- Even quite complex behavioral patterns requiring pattern
Examples of Behavior
A brief introduction to the fundamental units of behavior view, negative phototaxis would occur when the light im-
cannot begin to capture the remarkable range of behavioral pinging on one eye produced a strong ipsilateral motor
capacities known in animals. Various animals have evolved output (i.e., on the same side as the eye), causing the ani-
specialized sensory and motor capabilities that allow them mal to veer away from the source of the light. Positive pho-
to produce many interesting behaviors. We will consider totaxis would occur if the light stimulus to an eye stimu-
some well known examples to demonstrate the complexity lated contralateral locomotor output (i.e., on the side away
of the neuronal systems that underlie behavior: from the light), causing the animal to turn toward the
source of light. Consistent with this hypothesis, it has been
Animal orientation found that, if they are blinded in one eye, positively pho-
Many animals move predictably with respect to specific totactic animals will orient so that the intact eye points
stimuli-a behavior called orienting. This behavior re- away from the light.
quires the integration of sensory input and the coordination Sensory information modulates behavior in other ways
of motor output, so it depends on the properties of the sen- as well. For example, it can be used to correct for structural
sory receptor neurons, the connections within the central or functional asymmetries that arise in the central nervous
nervous system, and the muscles that cause the body of the system or in the structures (e.g., wings, legs, fins, etc.) that
animal to move. We now consider several examples of ori- influence locomotion. A locust can continue to fly in a
entation and navigation to illustrate some of the mecha- straight line even after one of its four wings is either partly
nisms that underlie these ubiquitous behaviors and to ex- or entirely removed, as long as it can use its eyes for orien-
emplify the complex behavioral capabilities of even simple tation. In the dark, even an intact locust that has been teth-
organisms. ered so that its behavior can be observed will roll about its
A common nocturnal sight for many city dwellers is longitudinal axis if it is induced to fly. The roll is due to
the scurrying of cockroaches as they abandon their snacks slight asymmetries in the structure of the wings and in the
and disappear when the kitchen light is switched on. This centrally generated motor output. A tethered locust ceases
behavior is an example of a taxis-a movement that is di- to roll if it is provided with visual cues in the form of an ar-
rected with respect to a stimulus. Scurrying cockroaches tificial horizon (see Figure 11-18B);visual input allows the
move away from a light stimulus, which is called negative motor output to the wings to be corrected, stabilizing the
phototaxis (Figure 11-18). An animal that turns toward insect's position. The visual input provides information to
the light is said to exhibit positive phototaxis. Jacques the central neurons that control flight, regulating the rela-
Loeb (1918) suggested early in the twentieth century that tive outputs to the left and right sets of wing muscles so
simple taxes occur when asymmetrical motor activation is as to hold the horizon seen by the animal in a horizontal
caused by asymmetrical sensory input. According to this orientation.
BEHAVIOR: I N I T I A T I O N , PATTERNS, A N D CONTROL 427
...............................................................................
The importance of sensory feedback for orientat~on orient toward potential victims that are as far away as
and locomotion in human beings is confirmed in our daily 0.5 m. At 15 cm or less, the scorpion can determine the dis-
experience. For example, as noted in Chapter 1,the driver tance, as well as the direction, of the source of vibration.
of a car continually makes minor steering adjustments: her Besides typical mechanoreceptor sensilla, a scorpion pos-
eyes are the sensors in a feedback system in which her neu- sesses a set of especially sensitive receptors on each of its
romuscular system, coupled to the car's steering mecha- eight legs (Figure 11-19).Although the sensitivity of these
nism, corrects any deviations that the car makes from the receptors to vibration is considerably less than that calcu-
center of the lane. Deviations may be due to asymmetries in lated for cochlear hair cells, it is nonetheless impressive.
the driver's nervous and muscular systems or to irregulari- With the use of a finely tuned mechanical displacement
ties or imperfections in the road and vehicle. When a per- stimulator, it has been shown that these receptors respond
son is deprived of this visual feedback, behavior changes. when the tarsal segment of the leg is displaced by less than
Blindfolded human subjects, either walking or driving a car 0.1 nm, which makes the receptors capable of detecting the
in an open flat field, take a more or less circular course, direction in which sand-borne vibrations are traveling.
with the size and direction (clockwiseor counterclockwise) To orient accurately toward the source of vibration, a
of the average circle differing among subjects. Similar turn- scorpion keeps all of its eight legs in contact with the sub-
ing biases are seen in animals at all phylogenic levels. Visual stratum in a precisely spaced circle (see Figure 11-19B). In
and other forms of sensory feedback compensate for these this arrangement, the leg closest to the source will be the
inherent locomotor biases, some of which may be due to first to sense the vibrational waves along the sandy sub-
congenital asymmetries in the function of the muscles and stratum. Central neurons that receive the sensory inputs
nervous system. from the receptors appear to compare the timing of im-
Many animals locate their prey from vibrations that are pulses that arise in the vibration receptors of all the legs.
set up in the substrate by their prey. A spider is alerted to Those legs facing the stimulus source intercept the waves
prey in its web, for example, by vibrations of the silken first, and those on the opposite side receive the stimu-
threads-vibrations that are detected by mechanoreceptor lus about 1ms later (vibrational waves travel 40-50 m spl .
organs located in the spider's legs. Another group of arach- in the sand). By integrating the timing of APs from the dif-
nids, the nocturnal desert scorpions, use sand-borne vibra- ferent legs, the central nervous system calculates the di-
tions produced by movements of their prey to locate and rection of the stimulus source, and it then produces the
I 5 cm
Gap
I I I I I I I
0.6 0.5 0.4 0.3
Time (s)
0.2
O1 t
Capture
Animal navigation been disabled. For example, it is now evident that, to vary-
Navigation over long distances is a striking extension of ing degrees, birds use particular landmarks, other visual
simple orienting responses. Many kinds of animals-from cues such as the plane of polarized light, odors, sounds,
monarch butterflies to golden plovers and gray whales- the position of the sun and stars, and even the Earth's mag-
migrate over long distances through unfamiliar territory. netic field as they find their way back home or to some
These navigational abilities have long been surrounded by other location.
an aura of mystery, because the animals typically rely on Animals from many phyla can navigate. Bees use the
cues that people cannot detect. In addition, the study of sun's position and the pattern of polarized light in the sky
navigation has been difficult because animals often use to keep track of the direction from their hive to food
several different sensory cues to guide them, and this re- sources, and they can signal this direction to their hive
dundancy has made the experimental study of navigation mates by their "waggle dances" (Figure 11-21).Some birds
confusing. Often, one system predominates when condi- can navigate over vast stretches of ocean that appear to be
tions for other systems are unfavorable; but, if conditions devoid of landmarks. When they are exposed to the night
change, the animal can shift its strategy. Experimenters- sky in a planetarium, some species of nocturnally migrat-
who are inclined to vary only one parameter at a time- ing birds-for example, warblers-orient with respect to
have found themselves mystified by an animal's continued the projected patterns of stars. As time passes during the
ability to find its way when one of its sensory modalities has night, if a projected constellation is moved across the dome
Figure 11-21 Bees can encode information
about the location of a food source with respect
t o the hive by indicating the direction of the
source related to the position of the sun. In this
I meridian
experiment, three potential food sources (num-
bered 1 to 3) were placed in very different loca-
tions. Scout bees that find a source indicate the
direction of the food with respect to the hive by
I Horizon - the direction in which they move on the vertical
wall of the hive. The direction ofthe straight part
of their "dance" shows the direction of the food
source relative t o the sun. [Adapted from
Camhi, 1984.1
vertical wall
of hive
"OW" I
to mimic the Earth's rotation, the birds orient themselves in The behavior of homing pigeons is a good example. Hom-
the "correct" direction with respect to the projected sky, ing pigeons deprived of familiar landmarks can still find
continually compensating for the time-dependent shift in their way home, even on an overcast day when it is impos-
the position of constellations. Arbitrary changes in the po- sible to detect the position of the sun. Under these condi-
sition of the projected sky will cause the birds to change tions, they typically fly in circles for a few minutes and then
their orientation. It appears that in order to compensate for head in the correct direction toward home. However, if
the rotation of the Earth relative to celestial references, small magnets are attached to their heads or if they are
birds, bees, and other celestial navigators refer to internal transported to release points in containers that make it im-
"clocks." This mechanism is called a clock-compass, and possible for them to detect Earth's magnetic field en route,
the underlying physiology is poorly understood. If a bee or they become disoriented. Local magnetic anomalies-for
a bird is put on a light-dark schedule with "dawn" and example, those caused by rich iron deposits-also disrupt
"dusk" shifted from natural lighting by several hours, it the pigeons' ability to orient properly.
will enter the incorrect time into its internal clock-compass Cave salamanders of the genus Eurycea can find their
and will orient with a compass deviation equivalent to the way "home" in complete darkness, so it seemed possible
artificial phase shift in its day-night cycle (Figure 11-22). that these animals, too, rely on Earth's magnetic field for
Animals can also depend on cues unavailable to human navigation. In experiments to test their ability to use a
beings to guide orientation. For example, although it has magnetic field as a navigational cue, salamanders were
long been suspected that some animals use the Earth's mag- trained to go to a particular place in a maze that was al-
netic field for orientation and navigation, only recently has ways oriented in the same way with respect to Earth's
the hypothesis been supported by experimental evidence. magnetic field. They were then tested in cross-shaped pas-
North netite in parts of the cerebral cortex. Catastrophic strand-
ings of these whales in unfamiliar inshore waters have been
shown to correlate significantly with the occurrence of
geomagnetic disturbances in the areas in which they swam
astray.
The distribution and location of magnetite suggests that
it may play a role in the detection of magnetic fields, but no
actual receptor cells that transduce magnetic signals into
West East neuronal signals have been identified. Some species of mud-
dwelling bacteria may provide a model showing how ori-
entation based on magnetite might work. These mud-
dwelling bacteria contain particles of magnetite, and they
have been found to orient to Earth's magnetic field. Bacte-
ria that live in the Northern Hemisphere orient toward
the North Pole, whereas those indigenous to the Southern
Hemisphere orient toward the South Pole, and this differ-
ence has been shown to depend on the orientation of
South
magnetite particles in the bacteria. The orientation of the
Figure 11-22 The clock-compass of homing pigeons can be manipu- magnetite enables the bacteria to swim downward into
lated by changing the timing ofthe pigeons' l~ght cycle. Homing pigeons deeper mud, following the angle with which the magnetic
were trained to return home from each of four locations.Then half of the
field enters the Earth at each location. If the bacteria are
pigeons were maintained on an artificial cycle that was 6 hours ahead of
the normal light-dark cycle. When the time-shifted pigeons were released placed in a drop of water within an artificial magnetic field,
from each of the test locations, they took off in a direction that was 90" they collect at the appropriate side of the water drop; if the
counterclockwise to the correct orientation. The solid black line indicates magnetic field is reversed, they swim to the opposite side of
the direction to the home cage. The solid red arrow indicates the aver- the drop.
age direction taken by pigeons that had been maintained on a normal
The American eel (Anguillayostrata) illustrates another
light-dark cycle, and the dashed red arrow indicates the average direc-
tion taken by pigeons that were time shifted. There were approximately method based On geomagnetism that is
10 pigeons maintained under each set of conditions. [Adapted from able only to marine organisms. Larval eels migrate from
Camhi, 1984.1 spawning grounds in the Sargasso Sea to the Atlantic coast
of North America, a distance of about 1000 km. When it
was initially suggested that they employ the Earth's mag-
sageways whose orientation to Earth's magnetic field could netic field for guidance, the response was skepticism be-
be manipulated. The results of the experiment confirmed cause the density of the magnetic field is so low. However,
that the salamanders can use Earth's magnetic field for the lateral-line system of eels contains extremely sensitive
orientation. This finding has significant implications con- electroreceptors, and these electroreceptors are the basis for
cerning the mechanism by which animals can detect mag- this ability. The movement of seawater in ocean currents
netic fields. Animals, such as birds, that move rapidly acts as an enormous generator, because the salt water func-
could be detecting the magnetic field directly. Alternatively, tions as a conductor moving through the Earth's magnetic
they could be detecting magnetic fields by sensing electri- field. The geoelectrical fields set up in the sea by ocean cur-
cal currents induced in their saline body fluids as these con- rents, such as the Gulf Stream, reach intensities of about
ductors are moved through Earth's magnetic field (as de- 0.5 mV.cm-l, or a 1.0 V drop over 20 km. The minute
scribed by Maxwell's laws of electricity and magnetism). electrical currents produced by these small voltage gradi-
Salamanders move very slowly compared with birds, and ents can apparently be detected by the eel's electroreceptors,
it is unlikely that electrical fields set up in their body flu- as shown by training eels to reduce their heart rates when
ids would be large enough to provide a basis for indirectly the surrounding electrical field changes. The heart rate of
detecting magnetic fields. It therefore seems most likely a trained eel drops when the surrounding electrical field
that at least some animals are able to detect magnetic fields changes by as little as 0.002 mV.cm-l. Because the fields
directly. generated in the ocean are two or three orders of magnitude
What sort of mechanism could underly a magnetic greater than this value, it is entirely possible that eels can
sense? This question cannot be answered with certainty. orient with respect to the geoelectrical field.
However, magnetite, a magnetic material of biological ori- This brief survey of animal orientation and navigation
gin, has been found in or near the brains of several animals reveals the complexity of the behavior generated by the un-
that appear to respond to magnetic fields-a finding that derlying neuronal systems. In view of these capabilities, it
is both intriguing and suggestive. For example, a small is daunting to begin a study of the neurons responsible for
black magnetite-containing structure lies between the producing behavior, and, as we will see, the descriptions of
brain and the skull of pigeons. Pelagic whales that may the neuronal basis of behavior commonly lag far behind the
navigate by using Earth's magnetic field also have mag- descriptions of the behavior itself.
PROPERTIES OF NEURONAL CIRCUITS
Despite the extreme complexity of nervous systems, several
generalizations can be made about their organization and
function. First, neuronal circuits consist of specific connec-
tions between neurons, and the pattern of connectivity is es-
sentially the same in all normal individuals of a species.
These connections, which are typically established during
embryonic development, are maintained and shaped by use
throughout an organism's life. If a particular circuit remains
unused for a long period, connections in the circuit become
weaker and there can be significant loss of function. Recent
evidence indicates that the opposite also is true: within cer-
tain limits, repeated activity can increase the strength of ex-
isting connections.
The crucial importance of proper connections in pro-
ducing behavior is illustrated by an experiment on a simple
reflex connection in a frog (Figure 11-23). In this experi-
ment, the consequences of altering neuronal connections
were tested by surgically disconnecting the sensory fibers
entering the spinal cord from one side and reconnecting
them to the dorsal root of the opposite side. The severed
frog neurons reconnected to the appropriate kind of central
neurons, but contacted the wrong side of the spinal cord. In
a normal frog, a noxious stimulus to a leg causes a reflex
withdrawal of the leg. After neuronal connections had re-
generated in this new configuration, a noxious stimulus to
the leg on the surgically manipulated side evoked an inap-
propriate withdrawal of the other leg. The abnormal con-
nections produced inappropriate behavior.
Connections within the nervous system also play an
enormously powerful role in sensory perception. In the
nineteenth century, Johannes Miiller formulated the law
of nerve-specific energy, stating that the modality of a
sensation is determined not by the nature of the stimulus,
but rather by the central connections of the nerve fibers
activated by a stimulus. This notion is now universally Figure 11-23 The "wiring d~agram"of the nervous system is crucial for
accepted, and we know that the various senses, as well producing appropriate behav~or.(A) In a normal frog, when a noxious
as the topographic distribution of sensory receptors, stimulus is applied to a leg, the leg is reflexly withdrawn. (B) Ifthe dorsal
are represented by particular regions of the cortex (see roots are cut and are caused to regrow into the contralateral spinal cord,
Figure 11-12).Direct electrical stimulation of local areas in the frog responds to a noxious stimulus on the surgically manipulated
side by withdrawing the opposite, unstimulatedleg. After the surgery, the
the somatosensory cortex evokes sensations in the subject's
sensory input from the right leg is connected to the motor neurons con-
consciousness that are more or less similar to those pro- trolling the left leg. [From "The Growth of Nerve Circuits" by R. W. Sperry.
duced by stimulation of the corresponding sense organ. Copyright 01959 by Scientific American, Inc. All rights reserved.]
Conversely, peripheral stimulation excites electrical signals
at points in the somatosensory cortex that correspond to
the specific region of the skin that was stimulated. nized into an enormously complex and varied circuitry. The
A second generalization about the nervous system is two basic kinds of signals-propagated, all-or-none APs
that the metabolic state, electrical properties, and summed and nonpropagated, graded synaptic and receptor poten-
total of all synaptic inputs impinging on each individual tials-are treated in Chapter 6. Synapses can be excitatory
neuron determine how that neuron will respond at any or inhibitory, strong or weak. In the end, all neuronal sig-
given moment. Each active neuron, by virtue of its connec- nals depend on the flow of ionic currents through ion chan-
tions, in turn influences activity in other neurons. nels, driven by electrochemical gradients.
A third generalization is that the complexity and vari- Behavior is produced when sensory information is re-
ety of functions carried out by nervous systems arise from ceived by the central nervous system, processed, and used
two levels of organization: (1)individual neurons can gen- to generate motor output. Alternatively, ongoing activity in
erate different kinds of signals and (2)neurons are orga- the central nervous system can, by itself, generate motor
BEHAVIOR: INITIATION, PATTERNS, A N D CONTROL 433,
...............................................................................
output-which happens, for example, when you decide to knee-jerk reflex) are independent of either a sensory filter
stop reading and close this book. To understand the entire network or a motor control network.
process, it is necessary to understand how the central ner- Even a small number of neurons can be combined into
vous system processes sensory information and how neu- circuits in a number of different ways. In fact, in higher ver-
ronal activity can lead to patterns of muscle contraction. tebrates, it is common for a single neuron to receive thou-
We now consider the pieces of a complete input-output sands of presynaptic terminals from other neurons, some of
pathway that can generate behavior: sensory input, central them excitatory and some inhibitory. Each neuron may it-
processing, and motor output. self branch many times and innervate many other neurons.
Divergence-the repeated branching of an axon-gives
Pieces of the Neuronal Puzzle each neuron widespread influence on many postsynaptic
In considering the complete network that underlies a neurons (Figure 11-24A).Convergence of inputs onto a sin-
particular behavior, we can recognize subcircuits whose gle neuron (Figure 11-24B)allows a neuron to integrate sig-
properties affect the way that the entire network functions. nals from many presynaptic neurons. Most neurons, such
Sensory filter networks transmit only certain features of as a mammalian spinal motor neuron, are rarely depolar-
complex sensory input, while blocking out other features. ized to threshold without considerable spatial and tempo-
Central pattern-generating networks (CPGs)produce pat- ral summation of excitatory synaptic input. As a result,
terned motor output that generates more or less stereo- neurons typically produce APs only when there is more or
typed movements. The output of some pattern genera- less simultaneous activity in a number of excitatory pre-
tors-for example, those governing locomotion and synaptic neurons, ~roducinga convergence of inputs. There
respiration-is cyclic. The output of other pattern gen- can be both divergence and convergence within a single net-
erators-such as those that control the movement of work. For example, when information from the retina is
the tongue when frogs and toads capture their insect transmitted to the brain, signals from each point at the cen-
prey-is noncyclic. Superimposed on some central pattern- ter of the visual field are carried in several neurons, indi-
generating networks is a motor command system in which cating divergence of information from single photorecep-
moment-to-moment changes in sensory input can modu- tors. In contrast, signals from the peripheral part of the
late the motor output. Some behaviors require the partici- visual field are combined over a wide area, indicating con-
pation of both a sensory filter network on the input side vergence of signals from many photoreceptors.
and a central pattern generator on the output side. An ex- Neurons typically receive simultaneous barrages of ex-
ample is the feeding reflex of the frog, described in the next citatory and inhibitory synaptic inputs, which are inte-
section. However, some of the simplest behaviors (e.g., the grated by the postsynaptic cell. Excitation of a neuron can
A D~vergence B Convergence
Figure 11-24 Information is carried through divergent and convergent neuron, allowing it to innervateseveral others. (B) Convergence refers to
pathways in the nervous system. (A) Divergence is the branching of one the innervation of a single neuron by many presynaptic neurons.
434 P H Y S I O L O G I C A L PROCESSES
.......................................
be suppressed if its active inhibitory synapses outweigh its
active excitatory synapses; so inhibitory synapses modulate
the ease with which excitatory inputs can bring the neuron
to threshold (Figure 11-25A). When many inhibitory
synapses onto a neuron are active, a greater number of ex-
citatory synapses must be activated to bring the post-
synaptic neuron to the firing level. However, the net effect
of inhibition depends on connections within the network
(Figure 11-25B).Inhibition of activity in an inhibitory neu-
ron-called disinhibition-can cause a net increase of ex-
citation in the network.
Feedback is employed extensively in neuronal cir-
cuits. An example of positive feedback is shown in
Figure 11-26A, in which a branch of a neuron in a hypo-
thetical reverberating circuit excites an interneuron that
feeds back to reexcite the initial neuron and keep it firing
for extended periods. In theory, this neuron could continue
to fire indefinitely once it has been excited by synaptic in-
put. If the interneuron in a feedback circuit is inhibitory in-
stead of excitatory (Figure 11-26B), it produces negative Figure 11-26 Neurons can be arranged in local feedbackcircuits. (A) Re-
feedback, reducing the probability that the initial neuron current facilitation. If the interneuron is excitatory, it produces positive
will fire. A well known instance of negative neuronal feed- feedback and prolongs activity in the initial neuron. (B) Recurrent ~nhibi-
tion. If the interneuron is inhibitory, it produces negative feedback and
back is that onto the motor neurons in the vertebrate spinal
limits activity in the initial neuron.
cord. Here, the a motor neurons make small branches that
innervate short inhibitory interneurons, the Renshaw cells
(Figure 11-27). Renshaw cells are excited each time the mo- tor neurons fire, and they feed back, reducing the level of
excitation within the motor neurons. The Renshaw cells re-
spond to stimulation by producing a high-frequency train
of impulses, which in turn cause inhibitory postsynaptic
potentials in the motor neurons. We do not fully under-
stand the functional significance of the Renshaw input
onto motor neurons, but we do know that it limits motor
neuron activity. Strychnine blocks the glycine-mediated
synapses made by Renshaw cells onto the motor neurons-
and probably other glycine-mediated synapses as well-
producing convulsions, spastic paralysis, and death from
paralysis of the respiratory muscles. These gruesome con-
sequences of blocking inhibitory synapses demonstrate the
importance of synaptic inhibition in normal function of the
nervous system.
Sensory Networks
Sensory neuronal networks-the first step in generating
appropriate behavioral responses-sort and refine the
mass of information that is available to an animal. Individ-
ual sensory neurons respond to only a limited range of
stimulus energy, which can be described by a tuning curve
(Spotlight 11-2). This property of receptors and other prop-
erties of sensory networks combine to filter incoming sen-
sory information. It is now clear that, in addition, sensory
neuronal networks can magnify, amplify, add, subtract, and
Figure 11-25 The net effect of inhibitory neuronal activity depends on even completely reconfigure the original pattern of sensory
the arrangement of the network. (A) Activity in the inhibitory neuron input. The visual system is particularly well studied, so we
(gray), reduces the probability of APs (indicated by a dashed arrow) in the
will examine it in some detail to illustrate some general or-
terminal cell, labeled T. (B)If two inhibitory neurons are arranged in series
and if the second one is tonically excited or spontaneously active, exci-
ganizational principles in sensory systems. In addition, we
tation of the first inhibitor will reduce inhibitory input onto the terminal will consider the auditory system of the barn owl as an ex-
cell (T), increasing its net activation (indicated by a solid arrow). ample of how neurons can transform sensory signals.
BEHAVIOR: INITIATION, PATTERNS, A N D CONTROL 435
...............................................................................
Figure 11-27 Renshaw cells feed back on and inhibit spinal u motor neu- shaw cell was excited by an antidromic AP caused in motor neuron b by
rons. A simplified diagram of motor neuron axons with their collateral an electrical stimulus. Action potentials in the Renshaw cell caused an in-
branches innervating a Renshaw cell. The Renshaw cells make glyciner- hibitory postsynaptic potential in motor neuron a. These inhibitory post-
gic inhibitory synapses onto the motor neurons. The trace shown next to synaptic potentials can be blocked by strychnine. [Adapted from Eccles,
each recording electrode illustrates a typical record. In this case, the Ren- 1969.1
separate objects. The neuronal mechanisms for emphasiz- What causes this illusion? It is a consequence of lateral
ing such differences probably evolved because small differ- inhibition at the level of the receptors. This phenomenon
ences in stimulus energy can be important cues to an ani- was studied in a series of experiments in the laboratory of
mal. You can observe contrast enhancement in your own H. K. Hartline at Rockefeller University in the mid-1950s,
visual system by looking at Figure 11-29. Each band in the and the work received recognition by a Nobel Prize in 1967.
figure appears to be lightest at its border with its darker In these experiments, the activity of a single ommatidium
neighbor and darkest at its border with its lighter neighbor. was recorded first when a bright stimulus light focused on
In fact, the actual luminosity of each band is uniform across only that ommatidium. You might suppose that adding
its entire width, and the apparent difference in brightness more stimulus light to the whole eye would increase the
within a band is an illusion. number of APs produced by that single ommatidium.
However, adding more light by switching on the room lights decreases with distance; inhibition is strongest between
decreased the firing rate of the single ommatidium. The dif- nearest neighbors. Lateral inhibition in the Limulus eye is
fuse light in the room stimulated the surrounding omma- mediated through the lateral plexus, a set of collateral
tidia and inhibited the test ommatidium. This phenomenon, branches from eccentric-cell axons that form inhibitory
lateral inhibition, has since been observed in many other vi- synapses on one another (see Chapter 7).Action potentials
sual systems, as well as in other sensory systems. in eccentric-cell collaterals cause the release of inhibitory
In another experiment, the source of lateral inhibition transmitter from synaptic terminals onto neighboring ec-
in the lateral eye of the horseshoe crab, Limulus, was centric-cell axons. Because the inhibition exerted by a unit
shown to be neighboring photoreceptors (Figure 11-30). on its neighbors increases as the unit's activity increases, a
While the response of a single ommatidium to a steady light strongly stimulated ommatidium will strongly inhibit
was recorded, a light pulse was presented to nearby om- neighboring, less strongly stimulated units. At the same
matidia. The onset of light in adjacent receptor cells caused time, the strongly stimulated unit receives weak inhibition
a reduction in the response of the ommatidium whose ac- from its neighbors. This interaction enhances the contrast
tivity was being recorded. Inhibition between interacting in activity level between neighboring units that are exposed
units is completely reciprocal, and the amount of inhibition to different intensities of light (Figure 11-31). Contrast
Direction of
movlng light
- 41C--
g 10 -
C
8, 8-
w
!?? I I I
LL
0 0.5 0 0.5
Distance between edge
and test ommatidium (mm)
Figure 11-31 Contrast is enhanced most strongly atthe boundaries be- ornmatidium (black plot in part 0). However, if the mask is removed, and
tween dim and bright regions. (A) In this experiment, the activity of a sin- the light border is allowed to pass over all the ommatidia, the output of
gle ommatidium is recorded, while a strongly lit rectangular f~eldis the single ommatidium will generate a response similar to that repre-
moved across the moderately lit compound eye. (B) Activity in the om- sented by the red plot. [From "How Cells Receive Stimuli" by W. H. Miller,
matidium is plotted againstthe position of the approaching bright edge. F. Ratliff, and H. K. Hartline. Copyright 01961 by ScientificAmerican, Inc.
If all other ommatidia are masked, the output of the ommatidium will All rights reserved.]
change in an abrupt, steplike manner as the bright rectangle reaches the
enhancement is greatest for units that lie at the boundary commonly faced by neuronal networks in general. In this
separating a bright and a dim region, because lateral inhi- section, we examine some of what is known about the neu-
bition diminishes with distance. Thus, lateral inhibition ronal processes underlying visual perception.
sharpens the visual detection of edges by increasing con- The visual pathway of vertebrates begins in the retina
trast at the borders between areas of different luminosities. and continues to the optic tectum in lower vertebrates
You experienced the effects of these interactions when you (Figure 11-32A), or to the lateral geniculate nuclei and
looked at Figure 11-29. the visual cortex in birds and mammals (Figure 11-32B).
Visual processing thus begins in the very first neurons The visual system can be viewed as a series of connected
of the network. Processing of visual information by neu- cellular plates (Figure 11-32C). The cells within each plate
rons farther along the chain continues to abstract and ac- have properties in common. In the projections from one
centuate properties of borders and other features of visual plate to the next, information both converges and diverges.
stimuli. There is a significant amount of synaptic interconnec-
tion and, hence, potential image processing within the
Information processing in the vertebrate retina retina itself. The photoreceptors connect with bipolar cells
The image of the world that falls onto the retina is a rela- that, in turn, contact ganglion cells, whose axons make up
tively accurate representation of the field viewed by that the optic nerve (Figure 11-33).The receptors are first-order
eye, limited only by the optics of the eye. The way in which cells, the bipolar cells are second-order cells, and the gan-
the visual system transforms this raw material into a per- glion cells are third-order cells in the afferent pathway. This
ceived image has been the subject of intense study at several nomenclature is oversimplified,because there are two other
levels of organization, ranging from the study of visual types of retinal neurons-the horizontal cells and the
transduction (see Chapter 7) to a consideration of neurons amacrine cells-that are particularly important for medi-
in the brain that might recognize entire perceived objects. ating lateral interactions within the retina. The horizontal
The visual system is the best studied of the sensory systems, cells receive input from neighboring and moderately distant
probably because vision is so important to primates, in- receptor cells, and they synapse onto bipolar cells. The
cluding human beings. However, principles that have amacrine cells interconnect bipolar cells and ganglion cells.
emerged from the study of vision apply to other sensory Studies combining intracellular recording techniques
systems as well, which suggests that evolution may have with the injection of fluorescent marker dyes have re-
settled on certain universal solutions to some problems vealed the electrical activity typical of each retina cell type
B Fields of vision
\ , , I
$ 8 ,
' ' I , # ,
,,> , :, ' '! ; , ,,'
s t
Optic tecturn
Amphibian
Visual cortex
Mammal
Figure 11-32 Visual information is transmitted from the retina to the tion are organized in layers. The retina contains the firstthree layers, and
brain through layers of cells. (A) In an amphibian, the left and right sides the remainder are in the brain, in the lateral geniculate nuclei and in the
of the optic tectum each receive projections from the entire field of view cortex. Information converges and diverges between the layers, and it
of the contralateral eye. (B) In a mammal, each side of the visual field is flows in both directions between the layers. [Part A from "Retinal Pro-
projected to the opposite side of the visual cortex. For example, the tem- cessing of Visual Images" by C. R. Michael. Copyright O 1969 by Scien-
poral half ofthe left retina and the nasal half of the right retina project to tific American, Inc. All rights reserved. Part B adapted from Noback and
the left visual cortex. (C)The neurons that initially process visual inforrna- Demarest, 1972.1
BEHAVIOR: I N I T I A T I O N , PATTERNS, A N D CONTROL 439
...............................................................................
1I
Outer
nuclear
layer
Figure 11-33 The function of the vertebrate retina is based on five ma- system through the optic nerves. Horizontal and amacrine cells, which are
jor types of neurons. Photoreceptors receive light stimuli and transduce located in the outer and inner plexiform layers, respectively,carry signals
them into neuronal signals. Bipolar cells carry signals from photorecep- laterally. [From "Visual Cells," by R. W. Young. Copyright O 1970 by Sci-
tors to the ganglion cells, which send their axons into the central nervous entific American, Inc. All rights reserved.]
(Figure 11-34). Vertebrate photoreceptor cells hyperpolar- in the center of the retina on which visual images are
ize when they are illuminated (see Chapter 7). They release sharply focused). This lack of convergence and divergence
synaptic transmitter continuously in the dark, and trans- produces very high visual acuity based on one-to-one-to-
mitter release is reduced when they hyperpolarize in re- one connections between cone photoreceptors, bipolar
sponse to illumination. Similarly, horizontal cells produce cells, and ganglion cells. (Cones are the majority of pho-
only graded hyperpolarizations in response to light (see toreceptors in the fovea.) Outside the fovea, each ganglion
Figure 11-34). Bipolar cells can produce graded potential cell receives input from many receptor cells-primarily
changes of either polarity. A ganglion cell responds with the rods-conferring on these ganglion cells a greater sensitiv-
' same polarity as do bipolar cells that innervate it. It be- ity to dim illumination but a lower degree of visual acuity.
comes depolarized and fires APs when the bipolar cells Structurally, the output of the retina is carried in the
synapsing on it depolarize, and it becomes hyperpolarized optic nerve by axons of ganglion cells, but how is the
and ceases spontaneous firing when its bipolar inputs hy- output organized? Understanding the information ex-
perpolarize. Amacrine cells respond transiently at the onset ported by ganglion cells hinges on the concept of a recep-
and offset of light in response to input from bipolar cells. tive field, an idea that was first proposed by Sherrington
Bipolar cells typically connect more than one receptor and was applied to visual processing by Hartline in the
to each ganglion cell, and they may also connect each re- 1940s. A cell's receptive field is the area on the retina in
ceptor cell to several ganglion cells. Thus, convergence and which light stimuli affect the cell's activity. The receptive
divergence already exist between the first- and third-order field of a ganglion cell is roughly centered on the cell and
cells of the visual system, but the amount depends on reti- varies in size, depending on the degree to which photore-
nal location. In mammals, both convergence and diver- ceptor and bipolar cells converge in the pathway to each
gence are minimal in the fovea, or area centralis, (the area ganglion cell. At the center of the fovea, a ganglion cell's
Figure 11-34 Each type of retinal neuron has a distinctive elec-
trical response to light. Activity was recorded in each type of cell
in response to a spot of light focused direaly on the receptors in
the region (left) and in response to an annulus of light surround-
v- -
ing the photoreceptors (right). The duration of the stimulus is in-
Spot 500 p m dicated in the lower trace on each record. In this example, the
annulus ganglion cell is activated by a light that shines on the center of its
receptive field. Note that the bipolar and ganglion cells produce
responses of opposite polarities to the spot and the annulus. This
effect is believed to be due to lateral inhibition similar to that in
RECEPTOR lntracellular record I 1 m~ Limulus. Notice that the off bipolar cell and the on-center gan-
glion cell shown in this figure would not be synaptically con-
nected. See Figure 11-36 for a detailed depiction of how ganglion
Light stimulus cell responses are related to signals in bipolar cells. [Adapted
from Werblin and Dowling, 1969.1
HORIZONTAL
v
OFF
BIPOLAR
ON-CENTER
I 2 m V
GANGLION
h-
receptive field extends over only one or a few photorecep- weaker off response is elicited by a spot of light that illu-
tors; at the periphery of the retina, where convergence is minates only a part of the field. The ring surrounding the
great, the receptive field of a ganglion cell can be as large as center of the receptive field is called the inhibitory surround
2 mm in diameter. of the receptive field. An off-center cell exhibits the con-
Each ganglion cell is spontaneously active in the dark, verse behavior, ceasing or reducing its activity when the
and the level of activity changes when a spot of light falls center of its receptive field is illuminated and increasing its
within its receptive field. Depending on which receptor cells firing when the surround is illuminated.
are illuminated, the frequency of APs in a ganglion cell may The center-surround organization of receptive fields de-
increase if a small spot of light enters the cell's receptive pends on lateral inhibition similar to that found in the com-
field-an on response. Alternatively the frequency of APs pound eye of Limulus. Lateral interaction in the vertebrate
may drop in response to a light-an off response. The re- retina takes place primarily through the activity of the hor-
ceptive field of a ganglion cell is typically divided into a cen- izontal cells in the outer plexiform layer (see Figure 11-33).
ter and a surround, and the response of the cell depends on The horizontal cells have extensive lateral processes and
whether the center or the surround is being stimulated or are interconnected with neighboring horizontal cells
both are (Figure 11-35). In an on-center ganglion cell, the through electrotonic junctions. In addition, they make
frequency of APs increases when the center of its receptive chemical synapses on bipolar cells and receive synaptic in-
field is illuminated (see Figure 11-35A). If a ring of light puts from receptor cells. Light that falls in the surround of
shines on the entire receptive field, with the center of the a ganglion cell's receptive field exerts its effects through lat-
ring over the center of the field, activity in the cell drops. A eral connections made by horizontal cells. Because hori-
A On-center ganglion cell B Off-center ganglion cell Figure 11-35 Retinal ganglion cells have ei-
Stimulus ther on-center responses or off-center re-
sponses to light stimuli. (A) Four recordings
I I I II
Stimulus Stimulus
zontal cells form an extensive syncytial network, commu- receptive field because it receives direct synaptic input from
nicating with one another through low-resistancegap junc- on bipolar cells. It is inhibited by light in the surround of its
tions, input from any receptor onto a horizontal cell pro- receptive field, because horizontal cells that receive input
duces a hyperpolarizing signal that spreads electrotonically from surrounding photoreceptors inhibit the on bipolar
in all directions away from the receptor. Every bipolar cell cells in the direct pathway from photoreceptors to the gan-
receives input from surrounding receptor cells by means of glion cell.
horizontal cells, and this input falls off with distance be- The responses of on and off bipolar cells depend on
cause the graded, hyperpolarizing potentials in horizontal how the cells respond to the neurotransmitter released by
cells decay as they spread electrotonically. The indirect photoreceptor cells and the different neurotransmitter re-
input that a bipolar cell receives from outlying receptors leased by horizontal cells. On bipolar cells are steadily hy-
through the horizontal cell network opposes the direct in- perpolarized in the dark by transmitter that is steadily se-
put that it receives from photoreceptors, providing the creted from the partially depolarized receptor cells. When
basis of the center-surround organization in retinal recep- a light stimulus causes photoreceptors to hyperpolarize,
tive fields. The local, direct-line pathway from photore- their release of transmitter drops and on bipolar cells are
ceptor through bipolar cell to ganglion cell produces the allowed to depolarize. This depolarization causes the on
center response. The indirect pathway from photorecep- bipolar cells to release an excitatory transmitter that depo-
tors through horizontal cells to bipolar cells and, thence, larizes ganglion cells, increasing the frequency of APs in the
to ganglion cells mediates the response to the surround. ganglion cells. In contrast, the off bipolars have a different
These two pathways show how particular features of a class of postsynaptic channels with different ionic selectiv-
stimulus can be extracted by even a relatively simple neu- ity and are steadily depolarized in the dark by neurotrans-
ronal network. mitter released by the photoreceptors. When light falls on
The distinctive responses of on-center and off-center the photoreceptors and they hyperpolarize, reduction in
ganglion cells arise from their connectionswith two classes their release of neurotransmitter causes the off bipolar
of bipolar cells: on bipolar cells and off bipolar cells. These cells to hyperpolarize. This hyperpolarization is accompa-
two types of bipolar cells respond oppositely to synaptic nied by a drop in transmitter release by the off bipolar
* input, both from receptors and from horizontal cells cells, producing a hyperpolarization of postsynaptic gan-
(Figure 11-36).The off bipolar cells become hyperpolar- glion cells.
ized by illumination of receptors, whereas the on bipolar In summary, the receptive field organization of the ver-
cells become depolarized. In both types of bipolar cells, a tebrate retina depends on three basic features:
light flashed onto the surround produces a response, me-
diated by horizontal cells, that is of the opposite electrical 1. Two kinds of ganglion cells receive input from
sign to that produced by illumination of the center. Each two corresponding kinds of bipolar cells. The connec-
bipolar cell causes potential changes in its ganglion cell or tions produce on-center and off-center ganglion cell
cells that are of the same sign as the potential change oc- responses.
curring in the bipolar. Thus, ganglion cells innervated by on 2. Receptors in the surround of the receptive field exert
bipolars will have on-center receptive fields, whereas those their effects through a network of electrically inter-
innervated by off bipolars will have off-center fields. An connected horizontal cells that synapse onto the two
on-center ganglion cell is excited by light in the center of its kinds of bipolar cells.
442 PHYSIOLOGICAL PROCESSES
..............................................................................
Optic nerve
Off center On surround
Figure 11-36 Connections within the retina produce the responses char- lar cells and ganglion cells to a spot of light. (B) Responses of bipolar cells
acteristic of on-center and off-center ganglion cells. Two kinds of bipo- and ganglion cells to an annulus of light. Amacrine cells have been omit-
lar cells, EON,and BoFF,respond oppositely to direct input from the re- ted from the diagram for simplicity. The direct pathway from photore-
ceptors, R, and to indirect input carried laterally by the horizontal cells, H. ceptors to ganglion cells, G, is shown in color. The indirect, lateral path-
The on bipolar cells become depolarized during activation of the overly- way through horizontal cells is shown in gray. The plus and minus signs
ing receptor cells and are weakly hyperpolarizedby lateral input from hor- indicate synaptic transfer that conserves (+) or inverts (-)the polarity of
izontal cells. The offbipolars behave oppositely. (A) Responses of bipo- the signal.
3. Direct input to bipolar cells from overlying receptors polarization of the presynaptic cell. The postsynaptic re-
and indirect input to the cells through the horizontal sponse depends on the ionic currents produced in the post-
cell network oppose each other and thereby produce synaptic cell as a result of the modulated release of trans-
the contrasting center-surround organization seen in mitter by the presynaptic neuron.
both the on-center and the off-center ganglion cells.
Information processing in the visual cortex
The organization of the retina reveals several general What happens to a retinal image after it has been trans-
principles that apply to the other parts of the central ner- formed into an array of receptive field responses within the
vous system. First, nerve cells can signal each other elec- retina? Physically, the information is carried by axons to vi-
trotonically without APs if the distances are small. Non- sual areas within the brain. The details of this pathway vary
spiking neurons can in fact convey more information more among species. In mammals and birds, the axons of retinal
accurately than can all-or-none signals. Electrotonic signals ganglion cells are routed either to the ipsilateral or to the
are attenuated with distance, which limits the range of ef- contralateral side of the brain at the optic chiasm, the site
fects such as lateral inhibition. Second, reception of stimuli where some axons cross the midline (see Figure 11-32B);
is not necessarily synonymous with depolarization. In some whereas, in vertebrates more primitive than birds, all op-
nerve cells (e.g., photoreceptors and some horizontal cells), tic fibers are routed to the contralateral side at the chiasm
hyperpolarization is the normal response to stimulation; it (see Figure 11-32A).To some extent, the degree of crossing
modulates synaptic transmission by causing a drop in the at the optic chiasm depends on how much overlap there is
steady release of transmitter. Third, the postsynaptic re- between the visual fields of the two eyes. In animals in
sponse in a neuron cannot be predicted from the sign of the which the visual field of one eye is entirely different from
potential change in the presynaptic neuron. A cell can be ei- the visual field of the other eye, all retinal ganglion cell ax-
ther depolarized or hyperpolarized in response to hyper- ons cross the midline. In mammals, ganglion cell axons
B E H A V I O R : I N I T I A T I O N , PATTERNS, A N D C O N T R O L 443
...............................................................................
synapse with fourth-order cells in the lateral geniculate nu- point in the visual field, but the eye of origin will switch
cleus of the thalamus. Lateral geniculate neurons send ax- from left to right as o w electrode moves from one layer to
ons to synapse on fifth-order cortical neurons in the occip- the next.
ital cortex (see Figure 11-11)in an area called Area 17, also Are there functional differences among the layers that
called primary visual cortex because it is the first region of receive information from each eye? Yes, the cells in each
the cortex in the pathway to receive visual information. layer respond to particular properties of a stimulus, and the
The pattern of synaptic relations within the lateral response varies from layer to layer. For example, in the
geniculate is based on the source and the nature of infor- monkey, cells within the four dorsal layers respond to the
mation carried by retinal ganglion cells, and it constitutes color of a stimulus, whereas cells of the two deepest layers
another step in the processing of visual input. Each lateral do not. In contrast, the two deepest layers respond to
geniculate nucleus, or body, is composed of six layers of movement, whereas the outer four layers do not. This spa-
cells, stacked like a club sandwich that has been folded tial sorting of outputs from ganglion cells illustrates an-
(Figure 11-37). The top four layers contain neurons with other principle of brain organization: information about a
small somata, which are called parvocellular neurons, and single stimulus is divided among parallel pathways. This
the bottom two layers contain neurons with large somata, pattern, called parallel processing, is a major theme of re-
called magnocellular neurons. Input onto these neurons is search into higher brain function. The receptive fields of
tightly organized. Each lateral geniculate body receives in- neurons in the geniculate do not differ substantially from
formation from only one half of the visual field (i.e., one of those of retinal ganglion cells. That is, they have a concen-
the two visual fields illustrated in Figure 11-32B), and cells trically arranged center-surround organization of either the
in each layer receive input from only one retina. Each neu- off-center or on-center type.
ron in the lateral geniculate receives information from only The difficult question of how the visual world is orga-
one eye. Neurons in a given layer receive information from nized in the next visual projection area, Area 17, was ex-
the same eye, and the layers alternate from one eye to the tensively and insightfully analyzed by David Hubel and
other, with the pattern of alternation changing between the Torsten Wiesel in the 1960s, and they received the Nobel
fourth and fifth layers (see Figure 11-37).Across all layers, Prize in 1981 in recognition of the importance of their
the topography of the corresponding retinal surface is pre- work. In their experiments, they recorded the activity
served exactly, and the topography is kept in register of individual neurons in the brain of an anesthetized cat
among the layers. If we pass an electrode along the path in- while a simple visual stimulus-such as a dot, circle, bar,
dicated by a dashed line in Figure 11-37, we will encounter or edge-was projected onto a screen positioned to cover
cells that respond to a light stimulus in precisely the same the entire visual field of the cat (Figure 11-38A). The
Ret~na cortex
Stimulus 1
responses that they recorded from cortical neurons were They found that cells of each type were arranged systemat-
correlated with the position, shape, and movement of the ically in space according to their optimal stimuli.
projected images. In retrospect, Hubel, Wiesel, and their The receptive fields of simple cells are long and bar
collaborators made two important decisions in their exper- shaped, and the on region of the field has a straight border
iments that allowed them to uncover order and regularity separating it from the off region (Figure 11-38B), rather
amid the enormous complexity of the visual brain. First, than the circular border found for cells in the retina and in
they chose to use more complex stimuli than just simple the lateral geniculate. As for retinal ganglion and geniculate
spots, and they asked which of these stimuli was most ef- cells, the receptive field of a simple cell lies in a fixed posi-
fective in eliciting a response in each neuron. Second, they tion on the retina and, hence, represents a particular part of
recorded from many cells with each electrode penetration, the total visual field. There is some variation in the recep-
allowing them to learn what neighboring cells had in com- tive fields of simple cells: some have a bar-shaped on region
mon and how cells were grouped in the brain. These strate- surrounded by an off region; for others, the receptive field
gies allowed them to discover several different kinds of or- consists of an off bar surrounded by an on region; and, for
der among the interconnections of the visual cortex, and still others, it consists of a straight edge with an off region
their discoveries have provided a model for examining on one side and an on region on the other side.
other sensory systems. A stimulus bar elicits maximal activity in a simple cell
Hubel and Wiesel's major discovery about the re- when it overlaps completely with the cell's on receptive field
sponses of cells in the visual cortex was that they responded (Figure 11-38C). When the bar is rotated so it no longer
to entirely different properties of stimuli than did retinal aligns with the orientation of the receptive field, either it has
ganglion cells. Cortical cells responded most strongly to no effect on the spontaneous activity of the simple cell or
bars projected in different orientations. They called the two it inhibits the cell's activity. If the bar of light is displaced so
major classes of cells that they found simple cells and com- that it falls just outside the on region, the cell is maximally
plex cells, based on the nature of their optimal stimulus. inhibited. The orientation and the on-off boundaries
BEHAVIOR: INITIATION, PATTERNS, A N D CONTROL 445
................................................................................
differ from one simple cell to another; so, when a bar of
light moves horizontally or vertically across the retina, it ac- On-center
tivates one simple cell after another as it enters one recep- retinal receptive
tive field after another. fields
I
What makes simple cells respond specifically to straight
bars or to borders of precise orientation and location?
Hubel and Wiesel suggested-and recent experiments have
I
confirmed-that each simple cell receives excitatory con- I Lateral
nections from lateral geniculate cells whose on centers are I geniculate cells
I
arranged linearly on the retina (Figure 11-39A). Simple I
cells that respond to borders, rather than to bars, are Receptive
field of
thought to receive inputs as shown in Figure 11-39B. A
simple cell
simple cell would receive maximal input when light fell on I
all of the receptors that activate the on-center fields of the L- - + Simple cell w i t h
ganglion cells and the geniculate cells on the pathway to bar-shaped fi.eld
that cell. Any additional illumination would fall on the in-
hibitory surround of the ganglion cells and could only re-
L!?
duce the response of the cortical cell.
Complex cells constitute the next level of abstraction in Off
the processing of visual information. Complex cells are be-
lieved to be innervated by simple cells, which would make
complex cells sixth-order cells in the hierarchy of visual in-
formation processing. Like simple cells, complex cells re-
spond best to straight borders of specific angular orienta- r-- Receptive fields
tion on the retina. Unlike the simple cells, however, I of lateral
I
complex cells do not have topographically fixed recep- I geniculate cells
I
tive fields. Appropriate stimuli presented within relatively I
large areas of the retina are equally effective at activating I
I
complex cells; as for simple cells, general illumination I
over the whole retina is not an effective stimulus. Some I
Recept~ve
complex cells respond to bars of light of specific orientation field of
(Figure 11-40A).Others give an on response to a straight simple cell
border when the light is on one side and an off response
when the light is on the other side. Still other complex cells
respond optimally to a moving border that progresses in
I
I
I
I
I
44 Lateral geniculate cells
or no response at all. These receptive fields can be explained on-off edge field
by a combination of synaptic inputs from simple cells. As a
light-dark border moves through the receptive fields of the
simple cells that synapse onto a complex cell, each simple
cell excites the complex cell in turn as the light-dark border
passes the in the Figure 11-39 The responses of simple cells in the visual cortex arise from
the simple cells. This arrangement could produce direc- the pattern of their synaptlc inputs. (A) The fixed, bar-shaped receptive
tional sensitivityto movement of the on-off boundary (see field of a s~mplecell arises from convergence of outputs from ganglion
~i~~~~1 1 - 4 0 ~ )1f. the boundary moved so that sequential and lateral geniculate cells whose circular on-center receptive fields are
linearly aligned. (B)An on-off straight-border receptive field results from
were One after the convergence of off-center and on-center geniculate cells onto the
would be excited, exciting the complex cell. When each simple cell.
simple cell became inhibited by the dark side of the moving
edge, the next one in line would be excited. In contrast, if
the boundary moved so that simple cells were sequentially a first step toward analysis and recognition. The spatial re-
exposed first to inhibition and only later to stimulation, one lations among visual cortical cells are correlated in an or-
simple cell after another would inhibit the complex cell, derly fashion with their functional properties. In their ex-
counteracting any tendency for excitation caused by the periments, Hubel and Wiesel discovered that cells adjacent
bright side of the edge. to one another responded to similar features of a stimulus.
The properties of individual cortical cells suggest that When they penetrated the visual cortex with an electrode
they abstract features of the visual scene, such as edges, as that was perpendicular to the cortical surface and recorded
446 PHYSIOLOGICAL PROCESSES
...............................
A
Rece~tivefields of
response response
Off On
"EdgeH-receptive
fields projected
from retina to
simple cells
cells
Figure 11-40 Responses in complexcells could be based on their pat- ity in the complex cell. In contrast, a horizontal bar of light produces only
tern of input from simple cells. (A) Some complex cells respond to bars a subthreshold response in the simple cells, and hence no signal is sent
of light that have a specific angular orientation, but their location can be to the complex cell. (B) Some complex cells respond to edges of light
anywhere within a large receptivefield. This pattern of response could be moving in only one direction. This response pattern could be produced
evoked by the convergence of many simple cells, each having similarly by the convergence of a population of simple cells, all of which are sen-
oriented bar-shaped receptive fields. In this example, the vertical bar of sitive to light-dark edges of the same orientation. Excitation of the com-
light stimulates one simple cell to fire because it falls on a row of gan- plexcell would occur ifthe edge were moved so that it illuminated the on
glion-cell receptive fields that produce the bar-shapedreceptive field of side of the simple-cell receptive fields before it illuminated the off side.
a simple cell. If the bar were moved to the right, it would excite another Movement in the opposite direction would produce only inhibition.
simple cell that synapses onto the same complex cell, producing activ-
responses from cells that were located along that pathway, sponding to touch or to the bending of a particular joint.
they discovered that the cells along each pathway re- However, the orderly array of orientation columns was
sponded to bars having the same orientation. When they only the first of the functionally based subdivisions found
moved the electrode laterally and made another penetra- in the visual cortex. The next to be discovered related to
tion, they found a column of cells that responded optimally the eye from which the visual signal came. By injecting one
to a stimulus having a different orientation from that of the eye with a radioactive tracer molecule that is transported to
optimal stimulus for the neighboring column of cells. Each the visual cortex, Hubel and his colleagues identified the
such set of cells is called a cortical column. In contrast, projection pattern of each eye onto the cortical surface.
recording from cells along a track parallel to the surface re- These experiments revealed a second columnar system in
vealed an amazingly regular change in the optimal stimulus which alternating columns represent one or the other eye
orientation, with the preferred orientation shifting about (Figure 11-41B, but see Spotlight 11-3).Three-dimensional
10 degrees each time the electrode advanced 50 pm. This reconstructions of these columns, called ocular dominance
result implies that the cells of the visual cortex are orga- columns, show their distribution across the cortical surface
nized in columns according to a feature of their optimal (Figure 11-41C).
stimulus and that this difference changes in an orderly fash- These experiments revealed that the visual cortex is
ion across the cortex (Figure 11-41A). subdivided into small functional units that analyze the stim-
The columnar organization of cells with similar re- ulus into its constituent features before passing it on to
sponse properties had been seen earlier in the somatosen- higher levels for further analysis. This modular organiza-
sory cortex, where adjacent columns contained cells re- tion is superimposed on a fundamental spatial map, which
BEHAVIOR: INITIATION, PATTERNS, AND CONTROL 447
...............................................................................
SPOTLIGHT 11-3 eyes from precisely the same small region of the visual field
These cort~calcells recelve extremely accurate neuronal projec-
SPECIFICITY OF tions from ganglion cells "seelngn the same part of the f~eld,but
located In the two retinas These f ~ n d ~ n g
conf~rm
s the suggest~on
NEURONAL of Johannes Muller, made more than a century ago, that Infor-
mation originating from analogous receptors (i.e., those that
CONNECTIONS AND
"see" the same part of the visual field) on both right and left reti-
INTERACTIONS nas converges on specific neurons in the brain. This high degree
of morphological specificity underscores the precision with
Notice in Figure 11-32B that the half of a visual image that falls which synaptic contacts are established within the central ner-
on the temporal part (the side toward the ear) of one retina falls vous system.
on the nasal part (the side toward the nose) of the other retina The neurons of the visual cortex are arranged in a remark-
and vice versa. In human beings, the ganglion cells on the right ably orderly manner.When a recording electrode is gradually ad-
side of each retina send their axons t o the right side of the brain, vanced through the cortex in a path perpendicular t o its surface,
and those on the left side send their axons t o the left side of the and encounters neurons in successively deeper layers, all of the
brain. Thus the nasal half of the right retina and the temporal half neurons along the track have receptive field properties in com-
of the left retina project t o the left side of the brain. mon. For example, all of the cells may b e simple cells with the
David Hubel and Torsten Wiesel, in their studies of visual same orientation.
processing in the brain, found that some neurons in the right and The precise and orderly arrangements of neuronal connec-
left visual cortices have receptive fields in both retinas, and that tivity present one of the greatest challenges t o neurobiology: t o
these receptive fields are optically in register. That is, cortical cells discover the mechanisms that guide neurons t o find functionally
that receive input from both retinas get information from both appropriate synaptic partners during embryonic development.
1Into cortex
Figure 11-41 Neurons of the visual cortex are arranged in columns that
are perpendicular to the cortical surface. (A) D~agramillustratingthe or-
ganization of columns of cells that respond to orientation of stimuli.
Columns are collections of cells for which the optimal stimulus orienta-
tion is the same for all cells in the column. Cells in adjacent columns have
different optimal stimulus orientations, and this orientation varies sys-
tematically from column to column. (B)The eye that excites cortical neu-
rons also alternates between neighboring columns. Red columns are ex-
cited by the left eye; gray columns by the right eye. (C) Simon LeVay's
reconstruction of the ocular-dominance columns in part of Area 17.
[Adaptedfrom Hubel, 1995.1
persists through the layers of the visual cells. To understand All of the various levels of cortical organization must
the nature of the spatial map at the cortical level, experi- be combined to provide the next set of cortical cells with
ments have been done to plot the visual field onto the a complete picture of visual stimuli, and the manner
cortex directly, with the use of a radioactive labeling tech- in which this synthesis is accomplished is still the subject
nique (Figure 11-42).Radioactive 2-deoxyglucose was in- of intense research. For example, it now appears poss-
jected into an anesthetized monkey and then a complex, ible that some high-order visual neurons may be active
target-shaped stimulus was projected onto its retina. Active only if a specific object (e.g., a face) enters their receptive
neurons take up more 2-deoxyglucose than do resting neu- field.
rons, so cortical neurons that were activated by the stimu- The visual cortex has taught physiologists several
lus would be expected to contain more radioactivity than important principles about the organization of sensory
their inactive neighbors. The pattern of radioactivity ob- networks. First, the visual system is organized hierarchi-
served in the visual cortex revealed that, although the two- cally. At each level, cells require more complicated stimuli
dimensional surface of the retina was completely repre- to excite them optimally, and this complexity arises from
sented on the cortical surface, the cortical pattern was not the convergence of cells having simpler receptive fields
an exact replica of the spatial features of the stimulus on the onto cells having more complicated receptive fields. Sec-
retina. Instead, retinal regions representing the center of ond, although convergence is apparent as we follow a
gaze (the fovea) were greatly magnified relative to those stimulus into the system, the parallel analysis of distinct
representing the peripheral view. This pattern corresponds features of a stimulus requires divergence of information
to the difference in visual acuity across the retinal surface, as well. The simultaneous analysis of different features of
as well as to differences in the convergence of primary pho- a stimulus, which occurs along parallel pathways, appears
toreceptors onto subsequent layers of neurons. This distor- to be an important principle of functional organization.
tion of the map in accord with the needs and habits of the Third, the activity of cortical neurons in Area 1 7 results
animal is characteristic of all animals with well developed in abstractions of some features of the visual stimuli.
visual systems. For example, animals, such as rabbits, that Fourth, the visual cortex does not receive a simple one-to-
live on large open plains have an elongated, horizontal re- one projection from the retina in either space or time. In-
gion of specialization, called a retinal streak, that provides stead, some regions in the visual field are expanded dra-
the greatest number of photoreceptors, and the least con- matically in their cortical representation, whereas others
vergence, for receiving stimuli along the visual horizon. are compressed.
Figure 11-44 Owls move thew heads to orlent toward a sound, a be-
havlorthat IS read~lyobservable (A) Experimental setup forstudylng the
ab~l~tyof an owl t o locate the source of a sound The target speaker can
be moved to any locatlon In a hemisphere surrounding the front of the
owl (6)Coordinate systems for determ~nlngthe locatlon of a sound El-
evatlon lnd~catesthe angle along a vertlcal axls and az~muthlndlcates the
angle In the horizontal plane Dlstance 1s malntatned constant [Adapted
from "The hear~ngof the barn owl" by E I Knudsen Copyr~ght0 1981
by Sclent~ficArnerlcan, Inc. All rlghts resewed.]
was plugged (Figure 11-4SA). With the right ear plugged, lies-at least in part-in anatomy. The region around the
the owl oriented below the actual source and slightly to the openings of an owl's ears is made of stiff feathers, called the
left. With its left ear plugged, it oriented above the source facial ruff,which form a surface that very effectively directs
and slightly to the right. In other words, when the sound sounds into the ear canals like the fleshy pinna of the mam-
was louder in the right ear, it seemed to the owl to be corn- malian ear. When these feathers are removed, the exter-
ing from above; whereas, when the sound was louder in the nal auditory canals of the owl are seen to be asymmetric
left ear, it seemed to be coming from below. The slight dif- (Figure 11-4SB).The opening of the right ear is directed up-
ference in the azimuth angle of orientation suggests that ward, whereas the opening of the left ear is directed down-
some information about the horizontal location is available ward. This arrangement could provide a basis for discrim-
from the intensity, but intensity cannot entirely account for inating elevation from intensity cues. The importance of the
orientation along that dimension. facial ruff was revealed by removing these feathers. If it
How can interaural intensity differences allow an owl lacked its facial ruff, an owl was no longer able to identify
to discriminate the elevation of a sound source?The answer the elevation of sound sources, although its estimates along
-
BEHAVIOR: INITIATION, PATTERNS, A N D CONTROL 451
...............................................................................
Preaural flap
/
Facial ruff
Figure 11-45 Plugging one of its ears caused an owl to make errors in site direction. (B) Facial ruff showing the asymmetry in the auditoryopen-
locating the source of a sound. (A)A target was presented directly in front ings. The right ear canal points slightly upward, whereas the left ear canal
of the owl, which had either a hard (high attenuation, solid symbols) or a points slightly downward. This small difference is amplified by the posi-
soft (less attenuation, open symbols) plug in one ear. Note that with the tion of the feathers in the facial ruff. [Adapted from "The hearing of the
left ear plugged (circles) the owl judged the sound to be above ~tsreal lo- barn owl" by E. I. Knudsen. Copyright 01981 by Scientific American, Inc.
cation. With the right ear plugged, the owl made mistakes in the oppo- All rights resewed.]
the horizontal axis remained as accurate without the facial sity of sounds arriving at each ear, and azimuth is judged
ruff as with it. Thus, the facial ruff must amplify the direc- from the ongoing temporal disparity between sounds ar-
tional asymmetry of the ears and is essential for discrimi- riving at each ear.
nating differences in elevation among sound sources. How is information about a sound's location in space
How does an owl locate sounds along the horizontal or represented in the nervous system? The ears cannot di-
azimuthal meridian? From behavioral experiments, it was rectly provide the brain with a representation of external
clear that differences in the time at which sounds arrived at space. Instead, as we have seen, an owl must compute the
each of the ears were important for this discrimination. difference in intensity between sound signals sensed by its
However, the relevant cue could have been either disparity two ears to determine the elevation of a sound, and it must
in the onset (or offset) of the sound or ongoing disparity compute an ongoing evaluation of disparity between
that occurred during the duration of sound (Figure 11-46). sound signals reaching its ears to determine the position of
Disparity of onset (or offset) refers to the difference in the the sound in the azimuthal plane. How and where these
time at which a given signal first reaches each ear; the ear comparisons are made and how the output is represented
nearest the source receives the signal first. Disparity can in the brain were discovered by Knudsen and Konishi in
' also occur between the signals that are received at the two the late 1970s.
ears as a sound continues; just as the onset of a sound Knudsen and Konishi identified a collection of space-
reaches the two ears at different times, identifiable features specific neurons in a midbrain nucleus. Each of these cells
of the sound reach first one ear and then the other. These responds best to sound signals that are located at a partic-
two types of disparity were independently varied by im- ular point in space, and each cell has a receptive field with
planting small speakers in an owl's ears. In response to dis- an on-center, off-surround organization similar to that
parity of onset, the owls failed to make "correct" head found in retinal ganglion cells (Figure 11-47A). Sounds that
movements; whereas, in response to ongoing disparities are located within the center region of the cell's receptive
ranging from 10 to 80 ps, owls made rapid head orienta- field (mean diameter = 25") excite the cell, whereas sounds
tions to the correct place in the azimuth corresponding to in the surround of the receptive field inhibit its response.
that time difference (Figure 11-46B). These experiments The neurons are arrayed in the nucleus to form a spatial
show that owls orient to sounds in space with remarkable map (Figure 11-47B) analogous to the retinotopic map de-
accuracy. Elevation is judged from differences in the inten- rived from the retina and the somatotopic map derived
452 P H Y S I O L O G I C A L PROCESSES
...............................................................................
Figure 11-46 Owls judge azimuthal location
from the disparity between ongoing sounds
that arrive at the ears. (A) Onset disparity oc-
curs when a sound reaches one ear before the
other. Ongoing disparity refers t o the contin-
ued difference in sound waves as perceived
by both ears. (B) Owls use the ongoing dis-
parity between the sounds reaching the two
ears t o localize a signal accurately in space.
The linear relation between azimuth and on-
going disparity between signals at the two
ears suggests that this type of disparity is the
relevant cue. [Adapted from "The hearing of
the barn owl" by E. I. Knudsen. Copyright O
1981 by Scientific American, Inc. All rights re-
Disparity Disparity sewed.]
at onset at offset
Right
ear
! I \ \ Ongoing disparity\ \ ! I
Left
ear
Time
Motor Networks
Optic
The sensory side of the nervous system acquires and ana-
tectum
lyzes information about the outside world, which is essen-
Left tial for producing behavior that is matched to an animal's
MLD current circumstances. This information must then be
passed on to neurons that are responsible for generating co-
ordinated movement. Relatively little detail is known about
Lateral
the interface between the sensory and motor sides of this
Posterior
3-Medial
Anterior
process, in part because investigators have worked inde-
pendently at understanding either sensory or motor sys-
tems. However, in a few cases, this sensory-to-motor con-
nection has been successfully explored, either in very simple
reflexes of vertebrates or in more complex behavior of
differences in sound intensity are computed by the owl's invertebrates.
brain are still being investigated. We will consider motor control systems of increasing
The barn owl's map of acoustic space was the &st ex- complexity, from those that produce simple reflex re-
ample of a brain map that is generated de nouo from the re- sponses through networks controlling repetitive actions to
454 P H Y S I O L O G I C A L PROCESSES
............................................................................
complex networks that reveal general principles of central
neuromotor organization. Motor patterns of different com-
plexity display various amounts of flexibility. Fixed action
patterns are relatively inflexible; they occur repeatedly with Central
little variation, but many behaviors are extremely plastic. feedback 1 I I
The animal can shape them to fit each new set of circum-
stances. One of the challenges for studies of motor control
is to understand the neuronal activity that allows an or-
ganism to produce behavior that changes from moment to
moment, as the situation changes.
muscles is the reflex arc. It takes only two kinds of neu- added to an isolated muscle or a joint bends, stretching the
rons-muscle stretch receptors (also called la-afferent muscle that runs over the joint), the muscle spindles are
neurons) and spinal a motor neurons-connected together stretched too. Stretching the central region of the muscle
to produce the myotatic reflex, or muscle stretch reflex spindles increases the frequency of APs in the la-afferent
(Figure 11-49A). Because the basic form of this reflex axons. These afferent axons make excitatory synapses di-
requires only the synapse between afferent and efferent rectly on the a motor neurons that control the muscle that
neurons, with no interposed interneurons, it is a monosyn- contains their spindle organs; so, when the activity in the
aptic reflex. la-afferent axons increases, it tends to excite the motor
BEHAVIOR: I N I T I A T I O N , PATTERNS, A N D CONTROL 455
...............................................................................
Figure 11-49 Only two klnds of neurons are requ~red
to produce the muscle stretch reflex (A) The steady
state of a muscle that 1s hold~ngup a l~ght we~ght(B)
If a heav~erwe~ght1s added to the muscle, ~tstretches
the muscle, actlvatlng stretch receptors, whlch
synapse onto or motor neurons In the same sp~nalseg-
ment and cause the muscle to contract more force-
fully If the sensory axons were cut, there would be no
l a afferent feedback onto the motor neurons and the we~ght
would cause the muscle to elongate (dashed Ihnes)
(C) Sequence of events that lead to produalon of the
stretch reflex.
Extra-
fusal
fibers
I
Error
Weight added Notice that, when a muscle contracts under the influ-
to muscle ence of the stretch reflex, tension is removed from the mus-
cle spindles. If nothing else happened, the la-afferents
$. Error would then become silent; and, if the muscle were to be
Length stretched a little more, the muscle spindles would be unable
to respond unless their own length could be regulated. The
intrafusal fibers-under the control of another set of mo-
tor neurons, the y-efferents-regulate the length of the
stretch receptors. When a muscle shortens, driven by its a
@ I I I I I I ~ I I I I I I lIl l l l l l i l l l l l l l laafferent motor neurons, activity in the y-efferents also causes the in-
APs in 1a afferents
trafusal fibers to shorten, maintaining a constant tension in
the spindle fibers. In this way, the y-efferents allow the spin-
@JlI I I I l l l l l ~l l LLl l l I l l l I I I I I I I L cv motor neuron dle fibers to maintain their sensitivity to muscle stretch
More APs In (Y motor neurons through a wide range of muscle length.
cause muscle to contract
Centrally generated motor rhythms
Locomotion and respiration typically consist of rhythmic
neurons, causing reflex contraction in the muscle that was movements produced by repetitive patterns of muscle con-
stretched (Figure 11-49B, C). traction. Each phase of such a neuromotor cycle is both
The stretch receptors provide negative feedback because preceded and followed by characteristic activity in motor
stretch of the muscle initiates neuronal activity that causes neurons. Bursts of activity are consistently related to one
the muscle to contract, opposing the stretch. A familiar ex- another in time. Logically, these repetitive acts could de-
ample of a stretch reflex is the knee-jerk reflex evoked when pend on moment-to-moment sensory input to the nervous
the tendon that crosses the knee cap (also called the patella) system or on the autonomous motor output of pattern-
is tapped. Tapping the tendon produces a sudden stretch of generating networks that happens entirely independently of
the quadriceps muscle on the ventral surface of the thigh, sensory input or on some combination of these two mech-
activating the muscle and causing the knee joint to extend. anisms (Figure 11-50).Regulation of repetitive motor out-
The arclike nature of the reflex is revealed when the dorsal put has been examined in many animal systems, and it ap-
root into the appropriate segment of spinal cord is cut. Sev- pears that both mechanisms play a role. These experiments
ering the dorsal root leaves all of the motor innervation in- are typically carried out in semi-intact preparations-that
tact but removes sensory input to the spinal segment. When is, in animals in which the nervous system has been exposed
the dorsal root is cut, the muscles innervated by the spinal for recording but that can still carry out recognizable
segment go limp, even though their motor input is intact. behaviors. In some behaviors, isolated nerve cords can
456 PHYSIOLOGICAL PROCESSES
...........................................
Figure 11-50 Motor output from the nervous sys-
tem depends on a comblnatlon of sensory lnput
Ongoing sensory
input I and central pattern generation Sensory Input
arlses In part from the environment and In part
from sensory receptors ~nthe body of the anlmal
Central pattern generators-represented by a
Pattern tape recorder In thls dlagram because these neu-
qenerator rons produce the same pattern of output over and
overagaln-play an Important role In shaplng be-
Sensory havlor, but they provlde only part of the Input onto
filter network the motor neurons.
Associative
networks
n
BEHAVIOR
produce all features of a motor output pattern; and, al- cles that cause alternate up-and-down movements of the
though the concept of an isolated nerve cord behaving may two pairs of wings, and these muscles receive the appropri-
seem strange, these behaviors can be studied either in semi- ate sequence of nerve impulses carried by several motor ax-
intact preparations or in isolated nerve cords, depending on ons. (See Chapter 10 for more information on insect flight.)
which is most convenient. The patterns of activity in these motor neurons continue to
Central motor patterns have been most clearly demon- occur with appropriate phase relations, even if sensory in-
strated in the nervous systems of some invertebrates-for put from the muscles or joints of the wings is eliminated by
example, in the neuromotor control of rhythmic locomo- cutting the sensory nerves (Figure 11-SlA).This persistence
tory movements. Grasshopper flight is controlled by mus- suggests that the motor pattern may be largely generated
A C Movement
\ CNS
/' 4
9
15
- (C)Cyclic organization of flight motor output. Externalsensory input (e.g.,
a puff of air on hair receptors) stimulates the flight motor output. Wing
movements activate stretch receptors that provlde Input stlmulatlng the
0 10 20 25 42 50 60 84 fllght motor Notlce that thls loop resembles the posltlve feedback loop
T ~ m e(s) illustrated In Flgure 11-26 [Adapted from Wllson, 1964,1971.]
BEHAVIOR: I N I T I A T I O N , PATTERNS, A N D CONTROL 457
.............................................................................
within the central nervous system by a network of neurons the ventral swim interneurons-which synapse onto the
that interact to coordinate the timing between contraction flexion neurons (Figure 11-52B).The cerebral neuron (C2),
of different muscles. the dorsal swim interneurons, and the ventral swim in-
Does sensory input play any role in the control of grass- terneurons are linked by reciprocal connections many of
hopper flight, which seems to be driven by centrally pro- which are a mixture of excitatory and inhibitory synapses.
grammed motor output? Sensory feedback from stretch re- Reciprocal inhibitory synapses between neurons have been
ceptors at the base of each wing is stimulated by the found in many central pattern generators that produce
movement of the wings and can modify motor output, in- rhythmic outputs; the reciprocal inhibitory synapses in the
creasing the frequency, intensity, and precision of the central pattern generator for swimming in Tritonia have
rhythm. If these receptors are destroyed, the neuronal out- been shown to be necessary for generating swimming in
put to the wing muscles slows down to about half its nor- this species. After the initial stimulus, the dorsal and ven-
mal frequency, although the phase relations among im- tral swim interneurons produce alternating bursts of neu-
pulses in the different motor neurons are retained. The ronal activity, which activate the flexion neurons responsi-
original frequency of the rhythm can be restored if the ble for motor output. Intracellular recordings of activity in
nerve roots containing the axons of the wing-joint recep- all five neuron types indicate that the swimming rhythm de-
tors are stimulated electrically (Figure 11-SIB). Interest- pends on both the membrane properties of individual neu-
ingly, although the motor rhythm increases in frequency rons and their synaptic connections. Thus, the rhythm is
when it is ~rovidedwith sensory input, the timing of the neurogenic, produced by interactions between neurons. Re-
motor output is not closely related to the timing of impulses cently, it has been demonstrated that synaptic strengths
in the sensory nerves. Random stimulation of the wing- among the neurons of this network can be modulated dur-
joint receptor axons can speed up the motor output, al- ing an episode of swimming to change the properties of the
though sensory input is most effective if it occurs during a network, even while it is producing the swimming output.
particular phase of the wingbeat cycle. Thus, propriocep- Autonomous central neuronal control also exists to
tive feedback is not required for proper phasing of motor various degrees in vertebrates. Respiratory movements,
impulses to the flight muscles; but, when the central flight which are driven by cells in the brain stem, persist in mam-
pattern generator becomes activated, sensory feedback re- mals when sensory input from the thoracic muscles is elim-
inforces its output (Figure 11-SIC). inated by cutting the appropriate sensory nerve roots.
What turns the flight motor on and off? When a Toads in which all sensory roots, except those of the cranial
grasshopper jumps off the substrate to begin flight, hair re- nerves, have been cut still produce simple coordinated
ceptors on its head are stimulated by the passing air. This walking movements, although these movements are hard
specific sensory input initiates the output of the flight mo- to discern, because loss of the myotatic reflex arc causes
tor. When the insect alights, the flight central pattern gen- muscles to become flaccid. Motor output to the swimming
erator is turned off by signals originating in mechanore- muscles of sharks and lampreys continues in a normal, al-
ceptors of the foot (the foot is called the tarsus in insects). ternating pattern when segmental sensory input is elimi-
Endogenous pattern-generating networks have been nated. However, the intersegmental sequencing of motor
shown to exist in a number of invertebrate nervous sys- output, which normally travels from the anterior to the
tems. For example, the cyclic motor output to the abdom- posterior may be disrupted.
inal swimmerets of the crayfish persists not only in an iso- Walking movements have been investigated in cats that
lated nerve cord, but even in single, isolated, abdominal are supported on a treadmill after the brain stem has been
ganglia. This intrinsic rhythm is initiated and maintained transected above the medulla oblongata (called a spinal cat
by activity of "command" interneurons, whose somata are preparation). Such studies reveal that the walking sequence
located in the supraesophageal ganglion of the brain. Al- can occur without input from the brain. Moreover, a rudi-
though the bursting pattern in each abdominal ganglion re- mentary walking rhythm has been seen to continue even af-
quires maintained activity in one, or perhaps several, of the ter the dorsal roots have been transected, eliminating sen-
interneurons, there is no simple one-to-one relation be- sory input. Thus, even in vertebrates, some aspects of
tween the firing pattern of these interneurons and the pat- rhythmic movements are programmed into intrinsic con-
tern of motor output to the swimmerets. The crucial in- nections between neurons within the spinal cord and hind
terneurons appear to be providing a general level of brain, and they can continue even if sensory feedback and
excitation, which keeps the central pattern generator active. other sensory inputs are disrupted.
One of the best studied rhythmic patterns is es-
cape swimming in the nudibranch mollusk Tritonia Central command systems
(Figure 11-52A). This sea slug swims away from noxious The stimulation of appropriate neurons in the central ner-
stimuli by making alternating dorsal and ventral flexions of vous system can elicit coordinated movements of various
its body, which are produced by alternating contractions of degrees of complexity. Electrical stimulation of one such
dorsal and ventral flexor muscles. The central pattern is command system, in the nerve cord of the crayfish, causes
generated by interconnections of three types of neurons- the animal to assume the defense posture, with open claws
a cerebral neuron (C2),the dorsal swim interneurons, and held high and body arched upward on extended forelegs.
CPG
Flexible
neurons
C2
DSI
DFN
Figure 11-52 Swimming in the nudibranch mollusk Tritonia is controlled properties and synaptic interactions determine the swim motor pattern,
by a central pattern generator consisting of three types of neurons. (A) If which changes if these parameters change. (C) Recordings of activity in
a Tritonia is threatened (such as by a nudibranch-eating starfish), it rises swim central pattern generating (CPG) neurons in an isolated brain after
off the substrate and swims by rhythmically contracting dorsal and ven- electrical stimulation of the pedal nerve.Abbreviations: C2, cerebral neu-
tral flexor muscles. (B) Three interconnected types of neurons act to- ron; DSI, dorsal swim interneurons; VSI, ventral swim interneurons; DFN,
gether to generate the swim motor pattern. An excitatory synapse is dorsal flexion neurons; VFN, ventral flexion neurons. [Part A courtesy of
represented by a bar; an inhibitory synapse by a solid circle; a combina- P. Katz; parts B and C adapted from Katz et al., 1994.1
tion of the two symbols represents a multifunctional synapse. Membrane
Appropriate sensory input excites this system through one the neuron is both necessary and suficient for causing the
specific interneuron, and this interneuron diverges broadly, particular motor output. That is, removing the neuron
producing excitation in some motor neurons and inhibition from the network must block, or greatly modify, the be-
in others. Command systems in arthropods characteristi- havior (necessity) and activating only that neuron must
cally activate many muscles in a coordinated manner and produce the behavior (sufficiency).
produce reciprocal actions in a given body segment; that is, When the necessity and sufficiency tests are carried
antagonists are inhibited while synergists are excited. Per- out to determine the neuronal basis of many behaviors,
haps not surprisingly, the command interneurons that are three observations appear again and again. First, many
most effective in eliciting a coordinated motor response are neurons are multifunctional, functioning differently under
generally least easily activated by simple sensory input. different conditions. For example, some retinal bipolar cells
The discovery of this command neuron in the crayfish have been found to carry signals from rod photoreceptors
initially caused physiologists to hypothesize that a lot of an in dim light and from cone photoreceptors in bright light.
animal's behavior might be controlled by a small popula- There must be a shift in their connectivity pattern as the
tion of command neurons, each of which was responsible level of ambient light changes. Second, one neuron may be-
for producing and shaping a particular behavior. In this long to different levels of a hierarchical control system (see
case, "choosing" among behaviors would depend on which Figure 11-48). For example, one neuron in the Tritonia
command neurons were most active. However, further swim control network acts both in the central pattern gen-
study of the neuronal basis of behavior suggests that most erator for swimming and in the command system for es-
command functions arise within networks of neurons, in cape. Third, because networks can be reconfigured, de-
which all contributing neurons play an important role. To pending on the situation, there must be mechanisms that
determine experimentally whether a neuron fills a com- can modify neuronal connectivity. Anatomical connections
mand function, it is necessary to show that the activity of may constrain the range of possible outputs for a set of neu-
BEHAVIOR: I N I T I A T I O N , PATTERNS, A N D CONTROL 459
............................................................................
rons, but functional connections define their output at any of these three principles in the organization of command
given time. One of the best understood mechanisms for systems.
shifting neuronal networks among possible functional con- Many invertebrates escape from potential predators by
figurations is neuromodulation (see Chapter 6 ) . Neuro- using stereotypical movements. One well studied example
modulators can cause changes in synaptic efficacy that dy- is the crayfish, Procambarus clarkii, which has two types of
namically reconfigure a collection of neurons into a new escape response, depending on the location of the stimulus
functional unit. Recognition that "anatomy is not destiny" (Figure 11-53A).In each behavior, at least one giant axon
in the nervous system has changed the way systems are an- is part of the control circuitry, a typical pattern in the neu-
alyzed. In this section, we consider two systems that have ronal control of many escape responses. Large axons carry
been analyzed in sufficient detail to provide examples signals rapidly, allowing an animal to escape faster. In the
Lateral giant
interneuron
Motor
Sensory
giant
receptor
Sensory
interneuron
Muscle I
Abdominal
segment
1, 2, o r 3
crayfish, there are two giant fibers: the medial giant in- circumstances. The premier example of dynamic network
terneuron, which controls flexion to propel the animal assembly can be found in a collection of 30 large neurons
backward; and the lateral giant interneuron, which plays that make up the stomatogastric ganglion (STG) of crus-
a key role in propelling the animal up and forward. The ba- taceans. The esophagus and stomach of lobsters and crabs
sic circuitry surrounding the lateral giant interneuron is is a complex structure that is responsible for ingesting, stor-
shown in Figure 11-53B. The neuronal network surround- ing, chewing, grinding, and filtering food (Figure 11-54).
ing the medial giant is quite different-on both the input There are four functional regions of the stomatogastric sys-
and the output sides of the network-which explains why tem: the esophagus, the cardiac sac, the gastric mill, and the
the behavior is so different when the crayfish is touched on pylorus. The neurons of the STG control all of the muscu-
its antenna. lar chambers responsible for ingestion and peristaltic move-
The escape responses of crayfish illustrate some other ment of food. They also control the bony teeth that are re-
features of motor control systems. First, if a crayfish is stim- sponsible for chewing and grinding it. Because most
ulated repeatedly, it fails to respond after about 10 minutes neurons in the STG are motor neurons that innervate the
of stimulation; the response is said to habituate. Although muscles in the stomatogastric system, their intrinsic prop-
habituation can occur at many different points in the net- erties have been of direct interest in efforts to discover the
work, it has been found that this behavior habituates be- functional architecture of each subnetwork that can be
cause less neurotransmitter is released from the terminals formed from this small set of neurons. The ganglion can be
of the sensory afferent neurons when stimuli are repeated divided into three networks of neurons, which control mus-
for long periods. Second, the overall control of the crayfish cles in the esophageal, gastric mill, and pyloric regions of
tail includes a second, parallel pathway that can also elicit the stomatogastric system. The esophageal, gastric, and py-
the tail-flip response. The fast flexor motor neurons, which loric networks can each generate patterns of rhythmic out-
are not giant motor neurons, produce more precise control put that are independent of the other two (Figure 11-55A).
of the tail flip, although it is neither as rapid nor as vigor- The frequency of output from each of the networks is a
1 ous as the flip produced by the giant neurons. When the tail
flip is initiated by the motor giant neurons, the second path-
characteristic of the network.
Input from modulatory neurons changes the behavior
way is activated, too, although the slow pathway can op- of these neurons drastically. For example, two electrically
erate by itself. Third, if the level of serotonin, a neuromod- coupled neurons, called PS neurons, reconfigure the net-
ulator, changes in the crayfish, the response of the crayfish works. When PS neurons fire, a valve between the esopha-
to a particular stimulus can change dramatically. An ag- gus and the stomach opens, and swallowing behavior is ini-
gressive crayfish can become submissive, and vice versa. tiated. Then an entirely new rhythm begins, coordinating
Thus neuromodulation must modify the connections be-
tween sensory and motor neurons.
The crayfish escape response is a typical fixed action
pattern, and the neurons that control it illustrate several
features of command systems outlined earlier. Perhaps the
most important feature is the existence of multiple control
points within the network, which offer several ways to ini-
tiate or to slightly alter the performance of the behavior.
This flexibility within the constraints of a fixed action pat-
tern has been a source of insight into the organization of
behavior.
dilator muscle
I enters
all three parts of the STG system to produce a set of peri- works that may be somewhat plastic, allowing flexibility in
staltic waves that travel from the esophagus to the pylorus behavioral responses. Understanding behavior at the neu-
(Figure 11-55B).All other rhythms are inhibited during this ronal level requires an understanding of how neurons in-
behavior. When activity in the PS neurons ceases, yet an- teract to produce behavioral output.
other rhythm transiently appears, but eventually all of the In the course of evolution, the primitive, distributed,
neurons in the swallowing network return to their original anatomically diffuse "nerve nets" characteristic of coelen-
activity patterns. The neurons that control swallowing be- terates became condensed into nerve cords and ganglia,
havior are called the swallowing network, which includes which are seen even in some jellyfish. In segmented animals,
neurons that, in the absence of PS neuron activity, are ac- the anterior end, initially specialized as the location of
tive in the esophageal, gastric, or pyloric networks. many sensory organs, became differentiated to contain a
Some of the neuromodulators that control the activity superganglion, or brain.
of STG neurons have been identified. The biogenic amine The most complicated nervous systems are found in
serotonin and the neuropeptides proctolin and cholecys- vertebrates. These systems can be divided into the central
tokinin all change the output pattern of at least some neu- and the peripheral nervous systems. All neurons in the ner-
rons in the STG. vous system are afferent neurons, efferent neurons, or in-
The reconfiguration of a small pool of neurons into sev- terneurons, and most of the neurons in complex neuronal
eral functional networks suggests a new view of the neu- networks are interneurons. To a large extent, the connec-
rons responsible for controlling motor output. Previous tions in central networks appear to be preprogrammed ge-
work has shown that a single anatomically defined network netically; but, during development and thereafter, they are
can produce different forms of output in response to neu- sustained by and can be modified by use.
romodulatory agents, but the stomatogastric system sug- The integration of input and the production of subse-
gests that the composition of the network, too, can be plas- quent activity in each neuron of a network depends pri-
tic. Dynamic specification of many functional networks marily on two major sets of factors: (1)the organization of
within a defined set of neurons offers a large increase in the circuits and synapses formed by interacting neurons, and
number of possible ways that motor output can be con- (2) the way in which individual neurons process or inte-
trolled. Clearly, one challenge is to discover where the con- grate incoming information to produce their own APs. The
trol of this mechanism resides and how it is regulated. integrative properties of a neuron depend on the anatomy
of the neuron, on its connections, and on the properties of
its cell membrane and ion channels.
SUMMARY The study of the neuronal control of behavior has been
All behavior is controlled by the motor output of the ner- aided by the identification of specialized behaviors called
vous system. Motor neurons are organized into &verse net- fixed action patterns. These highly stereotyped motor
462 PHYSIOLOGICAL PROCESSES
......................................
patterns are typically elicited by a particular stimulus, or system. How do they differ anatomically? How do
key stimulus. Understanding the behavioral capacities of they differ functionally and biochemically?
animals at the neuronal level is the goal of neuroethology. It has been said that all sensation takes place in the
Sensory neuronal networks sort and refine information brain. Explain what is meant by this statement.
available to the animal. They magnify, amplify, filter, and How can an increased rate of firing in an inhib-
reconfigure the original sensory input. The mammalian vi- itory interneuron produce increased activity in other
sual system has taught us a great deal about how sensory neurons?
systems function. Electrical recording from cells in the vi- What is the source of the continuous low-level synap-
sual cortex indicates that individual central neurons are ac- tic input and slow tonic firing in vertebrate spinal a
tivated by, and extract, certain features of a stimulus, rather motor neurons?
than generating a point-to-point representation of the pe- Describe the organization of the vertebrate retina.
ripheral input. In addition, studies of the visual system in- Each eye of a primate sees about the same field, but
dicate that there is a heirarchical arrangement of neurons the right hemisphere of the brain "sees" the left half
and that the specificity of sensory features evoking activity of the visual field, whereas the left hemisphere "sees"
in the neurons increases with each level, until only specific the right half of the field. How does this occur?
features of the visual stimulus evoke responses at the higher Why does the evening sky appear to have a lighter
levels. Some cells may be activated only by such compli- band outlining the silhouette of a mountain range?
cated stimuli as a face. Studies in the barn owl have shown What is meant by the "receptive field" of a cortical
that a map of auditory space is computed from intensity neuron?
and timing differences between sounds as they are received How can the receptive field of a simple cell in the vi-
at the two ears. Such computed spatial maps are in regis- sual cortex be a bar or a straight edge, when cells of
ter anatomically with other sensory maps, such as the the lateral geniculate have circular receptive fields?
retinotopic map from the visual system. What would happen to your posture if all of your
The simplest neuronal networks are monosynaptic re- muscle spindles suddenly ceased to function?
flex arcs, the most familiar of which is the stretch reflex of How do y-efferent fibers change the sensitivity of
vertebrates. More complex behaviors include locomotory muscle spindles?
movements that are based in part on central "motor pro- Discuss some of the general insights into neuronal or-
grams" that determine, for example, the sequence of mus- ganization that have resulted from studies of the
cle contractions that produce coordinated locomotory retina and visual cortex.
movements. Feedback from proprioceptive sensory neu- The nervous system is sometimes compared to a tele-
rons can exert an influence on the strength and the fre- phone system or a computer. Discuss some properties
quency of the motor output and, in most rhythmic motor of the nervous system that make this a good analogy
activity, it also contributes to fine-tuning its coordination. and others that make it a poor analogy.
Muscle output is controlled by a hierarchiacal system. What evidence indicates that some complex behavior
An example of the lowest level of control is the monosy- patterns are inherited and cannot be ascribed entirely
naptic stretch reflex arc that is responsible for maintaining to learning?
postural tone. At the next level are rhythmic motor patterns What supports the statement that the major factor re-
characteristic of walking, swimming, and crawling. Finally, sponsible for differences in the functioning of differ-
the control of complex fixed action patterns resides at the ent nervous systems is neuronal circuitry and not the
top of the hierarchy. Attempts to understand the relations properties of single nerve cells?
between levels of control have been most successful in the What are releasing stimuh and fixed action patterns?
relatively simple motor systems of invertebrates. In these Give at least one example of each.
model systems, it is clear that a particular neuron may par- What is a central pattern generator? What are some
ticipate in several motor networks that function at different of the properties of central pattern generators, and
levels. Moreover, in some systems, neuromodulatory sub- what roles do they play in the control of behavior?
stances dynamically control the configuration of networks. Describe some examples of central pattern generators.
What is a command neuron? What is a command
system? Describe some examples of command
REVIEW QUESTIONS
systems.
1. Action potentials in all neurons are fundamentally How can one neuron play different roles in several
alike; so how is the modality of input from the vari- central pattern generators?
ous sense organs recognized by the central nervous
system?
2. Describe the general organization of the vertebrate
SUGGESTED READINGS
brain and spinal cord. Camhi, J. 1984. Neuroethology. Sunderland, MA: Sinauer.
3. Compare and contrast the sympathetic and the (An excellent textbook summarizing many subfields of
parasympathetic divisions of the autonomic nervous this rapidly growing discipline.)
Carew, T. J., and C. L. Sahley. 1986. Invertebrate learning Kandel, E., J. Schwartz, and T. Jessell. 1991. Principles of
and memory: From behavior to molecule. Ann. Rev. Neural Science, 3d ed. New York: Elsevier. (A giant
Neurosci. 9:435-487. (A review of progress toward compendium of information about the nervous system,
understanding this important form of plasticity in the with some emphasis on vertebrate-particularly mam-
nervous system.) malian-species.)
Dowling, J. 1987. The Retina: An Approachable Part of Knudsen, E. I. 1981. The hearing of the barn owl. Scientific
the Brain. Cambridge, MA: Belknap Press. (A descrip- American 245:113 - 125. (A very readable discussion
tion of the structural and functional organization of the of the remarkable auditory nervous system of this bird,
vertebrate retina, written by a major contributor to our including a description of some very creative physio-
current knowledge of this remarkable organ.) logical experimentation.)
Ewert, J.-P. 1980. Neuroethology. Berlin: Springer-Verlag. Konishi, M. 1985. Birdsong: From behavior to neuron.
(An introduction to a relatively new field: the study of Ann. Rev. Neurosci. 8:125-170. (A review of the
the neuronal basis of behavior.) neuronal basis of the production of bird songs, written
Grillnel; S., and P. Wallen. 1985. Central pattern generators by one of the most eminent experts on the avian brain.)
for locomotion, with special reference to vertebrates. McFarland, D. 1993. Animal Behaviour: Psychobzology,
Ann. Rev. Neurosci. 8:233-261. (Areview of the prop- Ethology, and Evolution. New York: Wiley. (A classic
erties of central pattern generators, with emphasis on text covering the study o f animal behavior.)
the CPG for swimming in lampreys.) Nicholls, J. G., A. R. Martin, and B. G. Wallace. 1992.
Gwinner, E. 1986. Internal rhythms in bird migration. From Neuron to Brain: A Cellular and Molecular Ap-
Scientific American 25494 - 92. (A biological approach proach the Nervous System, 3d ed. Sunderland, MA:
to this otherwise apparently mysterious navigational Sinauer. (A very readable text describing both single-
ability.) cell properties and circuitry.)
Hubel, D. 1995. Eye, Brain, and Vision.New York: Scientific Nauta, W. J. H., and M. Feirtag. 1986. Fundamental
American Library Paperbacks. (Anexceedingly readable Neuroanatomy. New York: W. H. Freeman and Com-
review of information processing in the visual system writ- pany. (A comprehensive description of mammalian
ten by one of the most prolhc and creative researchers in neuroanatomy.)
the field.)
P A R T
INTEGRATION OF
PHYSIOLOGICAL SYSTEMS
c-
,
..
- ' - ~ o r s a l vessel
_,
/ /
~ u t Septum ~e'ntral Atera1 Nerve ANTERIOR
vessel heart cord
B Blood pressure
L0
Ventral vessel
Dorsal vessel
Time (s)
Figure 12-1 In the giant earthworm, Megascolides australis, peristaltic pumped into the ventral vessel. (B)Peak blood pressure is about twice as
contractions of the dorsal vessel and pumping by the lateral hearts are high in the dorsal vessel, because of its peristaltic contractions, than in
both important in moving blood. (A) Blood flows from the dorsal vessel the ventral vessel. [Adapted from Jones et al., 1994.1
into the lateral hearts, present in the 13 anterior segments, and then is
open circulation of two groups of invertebrates. The hemo- Animals with an open circulation generally have a lim-
coel is often large and may constitute 20%-40% of body ited ability to alter the velocity and distribution of blood
volume. In some crabs, for instance, blood volume is about flow. As a result, in bivalve mollusks and other animals
30% of body volume. Open circulatory systems have low that have an open circulation and use blood for gas trans-
pressures, with arterial pressures seldom exceeding 0.6- 1.3 port, changes in oxygen uptake are usually slow and max-
kilopascals (kPa),or 4.5-9.7 rnm Hg (1kPa = 7.5 rnrn Hg). imal rates of oxygen transfer low per unit weight.
Higher pressures have been recorded in portions of the open Nonetheless, such animals exert some control over both
circulation of the terrestrial snail Helix, but these are excep- the flow and distribution of hemolymph; moreover, the
tional. In snails, these high pressures are generated by con- blood is distributed throughout the tissues in many small
tractions of the heart, whereas in some bivalve mollusks high channels in animals with an open circulation. In the ab-
pressures in the foot are generated by contractions of sur- sence of such features, even moderate rates of oxygen con-
rounding muscles rather than of the heart. sumption would be impossible because of the large diffu-
CIRCULATION 469
......................................
A Hemocoel Ostium oxygen transfer. They have evolved a tracheal system in
Efferent \ with valves ,Heart which direct gas transport to tissues occurs through air-
filled tubes that bypass the blood, which plays a negligible
role in oxygen transport. Consequently, although insects
have an open circulation, they have a large capacity for aer-
obic metabolism. The insect tracheal system is described in
Chapter 13.
Closed Circulations
In a closed circulation, blood flows in a continuous circuit
of tubes from arteries to veins through capillaries. All ver-
Crayfish
\J' tebrates and some invertebrates, such as cephalopods
(octopuses, squids), have this type of circulation (Figure
12-2C). In general, there is a more complete separation of
functions in closed circulatory systems than in open ones.
Nephridial Mantle
B The blood volume in the closed circulation of vertebrates
vesse~s
typically is about 5% - 10% of body volume, much smaller
than that of open-circulation invertebrates. The extracellu-
lar volume in vertebrates, expressed as a percentage of
body volume is similar to the hemocoel volume in inverte-
brates. The closed circulatory system of vertebrates is a spe-
cialized portion of their extracellular space.
In a closed circulation, the heart is the main propulsive
organ, pumping blood into the arterial system and main-
taining a high blood pressure in the arteries. The arterial
system, in turn, acts as a pressure reservoir forcing blood
-. through the capillaries. The capillary walls are thin, thus
branchial vessel vessel branchial vessel
allowing high rates of transfer of material between blood
Bivalve mollusk and tissues by diffusion, transport, or filtration. Each tis-
sue has many capillaries, so that each cell is no more than
two or three cells away from a capillary. Capillary net-
works are in parallel, allowing fine control of blood distri-
Cephalic artery bution and, therefore, oxygen delivery to tissues. Animals
C A / with a closed circulation can increase oxygen delivery to
a tissue very rapidly. For this reason, squid, unlike many
other invertebrates, can swim rapidly and maintain high
rates of oxygen uptake; that is, their closed circulation
permits sufficient flow and efficient distribution of he-
molymph to the muscles to support short bursts of high-
level activity.
Vena cava ~ f f e r e n t Branchial
v
CePhalica branch~alartery heart
The blood is under sufficiently high pressure in a closed
circulation to permit the ultrafiltration of blood in the tis-
Cephalopod
sues, especially the kidneys. Ultrafiltration refers to the sep-
' Figure 12-2 Most invertebrates have an open circulation, but aration of an ultrafiltrate, devoid of colloidal particles, from
cephalopods have a closed circulation. The main blood vessels in the plasma by filtration though a semipermeable membrane
open circulationof crayfish (A) and bivalve mollusks (B) empty into a large (capillary wall) using pressure (blood pressure) to force the
surrounding space, the hemocoel, which makes up about 30% of total fluid through the membrane. Ultrafiltration occurs in most
body volume. Compared with an open circulation, the closed circulation
of cephalopods (C) is characterized by a higher blood pressure and more
vertebrate kidneys, resulting in the net movement of a pro-
efficient delivery of oxygen. In all diagrams only the main blood vessels tein-free plasma from the blood into the kidney tubule. In
are shown. The arrows indicate blood flow. general, all capillary walls are permeable and as pressures
are high, so fluid slowly filters across the walls and into the
space between cells. A lymphatic system has evolved in con-
junction with the high-pressure, closed circulatory system of
sion distances for oxygen between the hemolymph and the vertebrates to recover fluid lost to tissues from the blood.
active tissue. The extent of filtration depends largely on the blood pres-
Insects have an open circulation but do not depend on sure and the permeability of the capillary wall. Filtration
it for oxygen transport and thus can achieve high rates of across capillary walls can be decreased either by a reduction
470 INTEGRATION OF PHYSIOLOGICAL SYSTEMS
.........................................
in the permeability of the capillary walls or in the pressure of reservoir, and since there is little change in pressure as the
the blood. For example, the vessels in some tissues have less venous volume decreases, the volumes and flows in other
permeable walls than those found in other tissues. And in the regions of the circulation are not markedly altered.
liver and lung, where permeability is high for other reasons,
pressures are lower than those in the rest of the body.
THE HEART
Different pressures can be maintained in the systemic
(body)and pulmonary (lung)circulations of mammals be- Hearts are valved, muscular pumps that propel blood
cause the mammalian circulatory system is equipped with around the body. Hearts consist of one or more muscular
a completely divided heart (Figure 12-3).The right side of chambers connected in series and guarded by valves or, in
the heart pumps blood in the pulmonary circulation, and a few cases, sphincters (e.g., in some molluscan hearts),
the left side pumps blood in the systemic circulation. This which allow blood to flow in only one direction. The mam-
means, however, that the flows in the pulmonary and sys- malian heart has four chambers, two atria and two ventri-
temic circuits must be equal because blood returning from cles. Contractions of the heart result in the ejection of blood
the lung is pumped around the body. In other vertebrates into the circulatory system. Multiple heart chambers per-
the heart is not completely divided and flow to the lung can mit stepwise increases in pressure as blood passes from the
be varied independently of body blood flow. venous to the arterial side of the circulation (Figure 12-4).
The venous system collects blood from the capillaries
and delivers it to the heart via the veins, which are typically
low-pressure, compliant structures in which large changes
in volume have little effect on venous pressure. Thus, the
venous system contains most of the blood and acts as a
large-volume reservoir. Blood donors give blood from this
Neurogenic and myogenic pacemakers have been studied extensively in a variety of species. A myo-
In vertebrate hearts, the pacemaker is situated in the sinus veno- genic heart may contain many cells capable of pacemaker ac-
sus or in a remnant of it called the sinoatrial node (see tivity, but because all cardiac cells are electricallycoupled, the
Figure 12-4).The pacemaker consists of small, weakly con- cell (or group of cells) with the fastest intrinsic activity is the
trade, specialized muscle cells that are capable of spontaneous one that stimulates the whole heart to contract and deter-
activity.These cells may be either neurons, as in the neurogenic mines the heart rate. These pacemaker cells will normally
pacemaker in many invertebrate hearts, or muscle cells, as in overshadow those with slower pacemaker activity; however,
the myogenic pacemaker in vertebrate and some invertebrate if the normal pacemaker stops for some reason, the other
hearts. Hearts are often categorized by the type of pacemaker pacemaker cells take ovel; producing a new, lower heart rate.
and hence are called either neurogenic or myogenic hearts. Thus, cells with the capacity for spontaneous electrical ac-
In many invertebrates, it is not clear whether the pace- tivity may be categorized as pacemakers and latent pace-
maker is neurogenic or myogenic. Decapod crustaceans makers. In the event that a latent pacemaker becomes un-
(shrimps, lobsters, crabs), however, do have neurogenic coupled electrically from the pacemaker, it may beat and
hearts. In these animals, the cardiac ganglion, situated on control a portion of cardiac muscle, generally an entire cham-
the heart, acts as a pacemaker. If the cardiac ganglion is re- ber, at a rate different from that of the normal pacemaker.
moved, the heart stops beating, although the ganglion con- Such an ectopic pacemaker is dangerous because it will de-
k tinues to be active and shows intrinsic rhythmicity The car- synchronize the pumping action of the heart chambers.
diac ganglion consists of nine or more neurons (depending
on the species),divided into small and large cells. The small Cardiac pacemaker potentials
cells act as pacemakers and are connected to large follower An important characteristic of pacemaker cells is the absence
cells, which are all electrically coupled. Activity from the of a stable resting potential. Consequently, the membranes
small pacemaker cells is fed into and integrated by the large of cells in pacemaker tissue undergo a steady depolarization,
follower cells and then distributed to the heart muscle. The termed a pacemaker potential, during each diastole
crustacean cardiac ganglion is innervated by excitatory and (Figure 12-5).As the pacemaker potential brings the mem-
inhibitory nerves originating in the central nervous system brane to the threshold potential, it gives rise to an all-or-none
(CNS);these nerves can alter the rate of firing of the gan- cardiac action potential. The interval between cardiac APs,
glion and, therefore, the heart rate (beats per minute). which of course determines the heart rate, depends on the
Vertebrate, molluscan, and many other invertebrate rate of depolarization of the pacemaker potential, as well as
hearts are driven by myogenic pacemakers. These tissues the extent of repolarization and the threshold potential for
472 INTEGRATION OF PHYSIOLOGICAL SYSTEMS
............................................................................
Figure 12-5 Pacemaker cells undergo spon-
taneous depolarlzat~onof the membrane, re-
ferred to as the pacemaker potent~al,trlgger-
tng cardlac actlon potent~alsautorhymlcally
(curve A) A more rap~ddepolar~zat~on tn-
g (curve B ) and thus the
creases the f ~ r ~ nrate
heart rate, whereas a slower depolar~zat~on
slows the flrlng rate (curve C) and heart rate.
Dotential
Time (sec)
the cardiac Al? A faster depolarization brings the membrane other hand, norepinephrine released from sympathetic
to a firing levd sooner and thus increases the frequency of nerves accelerates the pacemaker depolarization potential,
firing, leading to a faster heart rate, whereas a slower depo- thus increasing the heart rate (Figure 12-6B). Although nor-
larization does the opposite (see Figure 12-5). epinephrine increases sodium and calcium conductance, this
Pacemaker activity has its origin in time-dependent probably is not the main mechanism involved in speeding up
changes in membrane conductance. In the frog sinus veno- the pacemaker rhythm. It is possible that norepinephrine de-
sus, the pacemaker depolarization begins immediately af- creases the time-dependent potassium efflux during diastole
ter the previous AP,when the potassium conductance of the and thereby increases the rate of pacemaker depolarization.
membrane is very high. The potassium conductance then
gradually drops, and the membrane shows a correspond- Cardiac action potentials
ing depolarization owing to the intracellular accumulation The action potentials that precede contraction in all verte-
of potassium ions and a moderately high, steady conduc- brate cardiac muscle cells are of longer duration than those
tance for sodium. The pacemaker depolarization continues in skeletal muscle. The AP in skeletal muscle is completed
until it activates the sodium conductance. The Hodgkin cy- and the membrane is in a nonrefractory stage before the on-
cle then predominates to produce the rapid regenerative up- set of contraction; hence, repetitive stimulation and tetanic
stroke of the cardiac AP (see Chapter 5). contraction are possible (Figure 12-7A).In cardiac muscle,
Acetylcholine, which is released from parasympathetic by contrast, the action potential plateaus and the mem-
terminals of the vagus nerve (tenth cranial nerve), slows the brane remains in a refractory state until the heart has re-
heart by increasing potassium conductance of the pacemaker turned to a relaxed state (Figure 12-7B).Thus, summation
cells. This increased conductance keeps the membrane po- of contractions cannot occur in cardiac muscle.
, tential near the potassium equilibrium potential for a longer Cardiac APs begin with a rapid depolarization that re-
time, thereby slowing the pacemaker depolarization and de- sults from a large and rapid increase in sodium con-
laying the onset of the next upstroke (Figure 12-6A). On the ductance. This differs from the slow depolarization of the
.-
E 0
Q)
+
g -20
0
C -40
2?
I
c2 Seconds
E -60 I
Q)
Stimulation
5
8
-80 Pacemaker
potential
+2? Seconds St~mulation
Pacemaker
potential
Figure 12-6 Parasympathet~cst~mulat~on vla the vagus nerve and sym- decrease In the durat~onand frequency of the actlon potentlal (B) Sym-
pathet~cst~mulat~on have opposite effects on the pacemaker potent~al pathet~cst~mulat~onproduces an Increase ~nthe frequency of flr~ngofthe
and heart rate (A) Vagus st~mulat~onproduces a rlse In dlastol~c(rest~ng) pacemaker cells [Hutter and Trautweln, 19561
transmembranepotent~al,a decrease In the rate of depolarlzat~on,and a
CIRCULATION 473
........................................
A Skeletal muscle B Cardiac muscle
I Plateau phase
Muscle contraction
Muscle contraction
Figure 12-7 Action potentials in skeletal muscle are of very short dura- st~mulation(0). For this reason, repetltlve stlmulatlon during a contrac-
tion (A), whereas cardiac action potentials exhibit a prolonged repolar- tion and summation of contractions can occur in the skeletal muscle but
ization, or plateau phase, during which cardiac muscle is refractory to not In heart muscle
pacemaker potential, which is marked by a stable sodium tricular AP is shorter, thus heart rates generally are higheq
conductance and decreasing potassium conductance. Re- than in larger mammals.
polarization of the cell membrane is delayed while the Because of the great diversity among the hearts of dif-
membrane remains depolarized in a so-called plateau phase ferent invertebrate phyla, few generalizations can be made
for hundreds of milliseconds (see Figure 12-7B).The long about the ionic mechanisms generating the cardiac AP of
duration of the cardiac AP produces a prolonged contrac- invertebrate hearts. The one widespread characteristic is
tion, so that an entire chamber can fully contract before any participation of calcium. For instance, bivalve mollusk
portion begins to relax-a process that is essential for ef- hearts have a calcium AP.
ficient pumping of blood.
The prolonged plateau of the cardiac AP results from Transmission of excitation over the heart
maintenance of a high calcium conductance and a delay in Electrical activity initiated in the pacemaker region is con-
the subsequent increase in potassium conductance (unlike ducted over the entire heart, depolarization in one cell re-
the situation in skeletal muscle). The high calcium conduc- sulting in the depolarization of neighboring cells by virtue
tance during the plateau phase allows Ca2+ions to flow of current flow through gap junctions (see Figure 4-35).
into the cell, because the equilibrium potential for calcium These junctions between cells are located in regions of close
is directed strongly inwards. This influx is especially im- apposition between neighboring myocardial cells, termed
portant in lower vertebrates, in which a considerable pro- the intercalated disk. Adhesion of cells at intercalated disks
portion of the calcium essential for activation of contrac- is strengthened by the presence of desmosomes. The area of
tion enters through the surface membrane. In birds and contact is increased by folding and interdigitation of mem-
mammals, the surface-to-volumeratio of the larger cardiac branes (Figure 12-9).The extent of infolding and interdig-
cells is too small to allow sufficient entry of calcium to fully itation increases during development of the heart and also
activate contraction. Therefore, most of the calcium is re- varies among species. Gap junctions are regions of low re-
leased-by depolarization of the T tubules (see Chapter sistance between cells and allow current flow from one cell
10)-from the extensive sarcoplasmic reticulum charac- to the next across intercalated disks.
teristic of the hearts of higher vertebrates. A rapid repolar- Although the junctions between myocardial cells can
ization terminates the plateau phase, due to a fall in calcium conduct in both directions, transmission is usually unidi-
conductance and an increase in potassium conductance. rectional because the impulse is initiated in and spreads
The duration of the plateau and the rates of de- only from the pacemaker region. There are usually several
polarization and repolarization vary in different cells of the pathways for excitation of any single cardiac muscle fiber,
same heart. The summation of these changes are recorded since intercellular connections are numerous. If a portion
as the electrocardiogram (Figure 12-8).Atrial cells gener- of the heart becomes nonfunctional, the wave of excitation
ally have an AP of shorter duration than ventricular cells. can easily flow around that portion, so that the remainder
The duration of the AP in atrial or ventricular fibers from of the heart can still be excited. The prolonged nature of
hearts of different species also varies. The duration of the cardiac APs ensures that multiple connections do not result
AP is one factor correlated with the maximum frequency of in multiple stimulation and a reverberation of activity in
the heartbeat; in smaller mammals, the duration of the ven- cardiac muscle. An AP initiated in the pacemaker region
474 INTEGRATION OF PHYSIOLOGICAL SYSTEMS
. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
A Electrocardiogram Nucleus Myocardial cells
I / \
intercalated disk;
B Cardiac action potentials
Pacemaker h Figure 12-9 Electrical activitycan spread throughout the heart because
of the close apposition of myocardial cells at intercalated disks, which are
densely packed with gap junctions. Shown here is a schematic diagram
of myocardial cells from a mammalIan heart. The folding and the inter-
digitation of membranes are characteristicof intercalateddisks. Although
desmosomes are present in these reglons, thereby strengtheningthe ad-
hesion between cells, they are not easily distinguished.
Increasing
catecholamine
/ stimulation
The end-systolic volume is determined by two parameters: elevated arterial pressure results from the increased stretch
of cardiac muscle during diastole.
The pressures generated during ventricular systole As noted above, epinephrine and norepinephrine re-
The pressure in the outflow channel from the heart leased from sympathetic nerves or circulating in the blood
(aortic or pulmonary artery pressure) increase the force of contraction of the ventricle; hence both
the rate and the extent of ventricular emptying are in-
Increasing the venous filling pressure causes an increase in creased by these catecholamines. The effects of cholinergic
end-diastolicvolume and results in an increased stroke vol- (i.e., vagus) nerve activity on the rate and force of ventric-
ume from an isolated mammalian heart (Spotlight 12-1). ular power output during each beat are much less marked
End-systolic volume also increases, but not as much as end- than the effects of adrenergic sympathetic nerves. This dif-
diastolic volume. Thus, cardiac muscle behaves in a way ference stems from the much more extensive innervation of
similar to that of skeletal muscle in that stretch of the re- the ventricles by adrenergic nerves than by cholinergic
laxed muscle within a certain range of lengths results in the nerves.
development of increased tension during a contraction. In- The effects of sympathetic nerve stimulation andlor in-
creases in arterial pressure also cause a rise in both end- creased circulating levels of catecholamines represent a se-
diastolic and end-systolic volume with little change in ries of integrated actions. Stimulation of pacemaker cells
stroke volume. In this instance, the increased mechanical leads to an increase in heart rate. Conduction velocity over
work required to maintain stroke volume in the face of an the heart is increased to produce a more nearly synchro-
CIRCULATION 477
......................................
nous beat of the ventricle. Both the rate of production of but in this instance there are large changes in stroke volume
ATP and the rate of conversion of chemical energy to me- rather than in heart rate. The increases in cardiac output
chanical energy in ventricular cells increase, leading to an are probably caused by an increase in venous return. The
increase in ventricular work: the rate of ventricular emp- sympathetic nerves are not involved in increasing cardiac
tying increases during systole, so that the same or a larger output per se but rather in raising heart rate and maintain-
stroke volume is ejected in a much shorter time. This in- ing stroke volume, avoiding the large pressure oscillations
creased force of contraction is mediated by the action of associated with large stroke volumes and keeping the heart
catecholamines on both a- and p-adrenoreceptors (see operating at or near its optimal stroke volume for efficiency
Chapter 8 for more details). Thus, when adrenergic nerve of contraction. The sympathetic nerves thus play an im-
stimulation increases the heart rate, the same stroke volume portant role in determining the relation between heart rate
is ejected from the heart in a shorter time. So, although the and stroke volume, but additional factors are involved in
time available for emptying and filling the heart is reduced mediating the increase in cardiac output with exercise.
as heart rate increases, stroke volume remains quite stable
over a wide range of heart rates. For example, exercise in Changes in pressure and flow during a single heartbeat
many mammals is associated with a large increase in heart Contractions of the heart cause fluctuations in cardiac
rate with little change in stroke volume; only at the high- pressure and volume as illustrated by the tracings in
est heart rates does the stroke volume fall (Figure 12-10). Figure 12-11A. The following sequence of events occurs
This situation occurs because, over a wide range of heart during contraction of a mammalian heart (Figure 12-11B):
rates, increased sympathetic activity ensures more rapid
ventricular emptying, and elevated venous pressures result 1. During diastole closed aortic valves maintain large
in more rapid filling as heart rate increases. pressure differences between the relaxed ventricles and
There are limits, however, to which diastole can be the systemic and pulmonary aortas. The atrioventric-
shortened, determined by the maximum possible rate at ular valves are open, and blood flows directly from the
which the ventricles can be filled and emptied as well as the venous system into the ventricles.
nature of the coronary circulation. During contractions of 2. When the atria contract, pressures rise in them and
the myocardium, coronary capillaries are occluded, so flow blood is ejected from them into the ventricles.
is very reduced during systole. Flow rises dramatically dur- 3. As the ventricles begin to contract, pressures rise in
ing diastole, but a decrease in the diastolic period tends to them and exceed those in the atria. At this point, the
reduce the period of coronary blood flow. Catecholamines atrioventricular valves close, thus preventing backflow
also cause coronary vasodilation and increase coronary of blood into the atria, and ventricular contraction
flow. As we have noted, increases in cardiac output with ex- proceeds. During this phase, both the atrioventricular
ercise often are associated with large changes in heart rate and the aortic valves are closed, so that the ventricles
and small changes in stroke volume in mammals (see form sealed chambers and there is no volume change.
Figure 12-10).Following sympathetic denervation of the That is, the ventricular contraction is isometrzc.
heart, exercise results in similar increases in cardiac output, 4. Pressures within the ventricles increase rapidly and
eventually exceed those in the systemic and pulmonary
aortas. The aortic valves then open, and blood is
Maximum O2 ejected into the aortas, resulting in a decrease in ven-
consumption
I tricular volume.
I
Cardiac
I 5. As the ventricles begin to relax, intraventricular pres-
sures fall below the pressures in the aortas, the aortic
valves close, and there is an isometric relaxation of the
ventricle.
Figure 12-11 During a single cardiac cycle, sequential contraction of the ing a single cardiac cycle. (B) Sequence of events in contraction of mam-
atria and ventricles and the opening and closing of valves produce char- malian heart. Black indicates contracted muscle; gray, relaxed muscle.
acteristic changes in pressure and volume. (A) Changes in pressure and See text for discussion. [Part A adapted from Vander et al., 1975.1
volume in the ventricles and aorta (left) and pulmonary artery (rightjdur-
tension in the muscle and pressure in the ventricle increase ventricles eject equal volumes of blood, but the pressures
rapidly. The second phase is essentially isotonic; tension generated in the pulmonary circuit (right ventricle) are
does not change very much, for as soon as the aortic valves much lower; consequently, the external work done by the
open, blood is ejected rapidly from the ventricles into the right ventricle is much less than that done by the left ven-
arterial system with little increase in ventricular pressure. tricle. As described in the previous section, blood is ejected
Thus, tension is generated first with almost no change in from the ventricle when intraventricular pressures exceed
length; then the muscle shortens with little change in ten- the arterial pressure. If the arterial pressure is elevated,
sion. In other words, during each contraction, cardiac mus- more external work must be done by the heart to raise the
cle switches from an isometric to an isotonic contraction. intraventricular pressure enough to maintain stroke volume
at the original level. This, of course, means that there is an
Work done by the heart extra strain on the heart if blood pressure is high.
It is a simple principle of physics that external work done is Not all energy expended by the heart will appear as
the product of mass times distance moved. In the present changes in pressure and flow; some energy is expended to
context, work can be calculated as the change in pressure overcome frictional forces within the myocardium, and
times flow. Flow is directly related to the change in volume more is dissipated as heat. The external work done by the
with each beat of the ventricle. With the pressure given in heart, expressed as a fraction of the total energy expended,
grams per square centimeter and the volume in cubic cen- is termed the efliciency of contraction. The external work
timeters, pressure times volume equals grams times cubic done can be determined from measurements of pressure and
centimeters divided by square centimeters, which equals flow and converted into milliliters of 0, consumed. This, in
grams times centimeters-the equivalent of mass times dis- turn, can be expressed as a fraction of the total 0, uptake
tance moved, or work. Thus, a plot of pressure times vol- by the heart in order to measure the efficiency of contrac-
ume for a single contraction of a ventricle yields a pressure- tion. In fact, not more than 10% -15% of the total energy
volume loop whose area is proportional to the external expended by the heart appears as mechanical work.
work done by that ventricle. Energy is expended to increase wall tension and raise
Figure 12-12 illustrates pressure-volume loops for the blood pressure within the heart. According to Laplace's
right and left ventricles of a mammalian heart. The two law, the relationship between wall tension and pressure in
CIRCULATION 479
...............................................................................
pericardial sac depend on the rigidity of the pericardium
and on the magnitude and rate of change of the heart vol-
ume. The membrane may be thin and flexible (compliant),
in which case pressure changes within the pericardial cav-
ity during each heartbeat are negligible. Or the pericardium
may be quite rigid (noncompliant), in which case the in-
trapericardial pressure oscillates during each heartbeat.
The compliant pericardium enveloping the mammalian
heart is formed of two layers, an outer fibrous layer and an
inner serous layer. The serous layer is double, forming the
inner lining of the pericardial space and the outer layer (epi-
cardium) of the heart itself. In mammals, the serous layer
secretes a fluid that acts as a lubricant, facilitating move-
ment of the heart.
Crustaceans and bivalve mollusks have a noncompliant
pericardium. In these animals, contractions of the ventricle
reduce pressure in the pericardial cavity and enhance flow
I
+
0 50
Filling
100
+
150 200
into the atria from the venous system (Figure 12-13).Thus,
tension generated in the ventricular wall is utilized both to
Volume (ml) eject blood into the arterial system and to draw blood into
the atria from the venous system.
Figure 12-12 The area of a ventricular pressure-volume loop is propor-
The peficardium of elasmobranchs (sharks)and lung-
tional tothe external work done by a ventricle in one cardiac cycle. Shown
here are loops for the right and left ventricles of the mammalian heart.
fishes also is noncompliant, whereas that of teleosts is com-
Once around a loop in a counterclockwise direction is equivalent to one pliant. The elasmobranch heart consists of four cham-
heartbeat. Ventricular filling occurs at low pressure; pressure increases bers-sinus venosum, atrium, ventricle, and conus-all
sharply only when theventricles contract (the sharp upswing on the right- contained within a rigid pericardium (Figure 12-14).The
hand side of each loop). Ventricularvolumedecreases as blood flows into reduction in intrapericardial pressure that occurs during
the arterial system, and ventricular pressure falls rapidly as the ventricle
relaxes. Filling then begins again. Note that although the volume
ventricular contraction in elasmobranchs produces a suc-
changes in both ventricles are similar, the pressure changes are much tion that helps expand the atrium and thereby increases ve-
larger in the left ventricle than in the right one. Therefore, the left ventri- nous return to the heart. If the pericardial cavity is opened,
cle has a larger loop and hence does more external work than the right cardiac output is reduced; hence the increased venous re-
ventricle.
turn to the atrium caused by reduced pericardial pressure is
Afferent Ventral
branchial aorta Ventricle
artery
Figure 12-15 In a "typical" water-breathingteleost such as the trout, the aorta. Most teleosts contain a low-hematocrit secondary circulation,
respiratory circulation through the gills and the systemic circulation are in which supplies nutrients but not much oxygen to the skin and gut. Black
series. In the four-chamber, undivided heart, the pacemaker is in the si- arrows indicate flow of deoxygenated blood; red arrows, flow of oxy-
nus venosus. The ventricle ejects blood into the compliant bulbus and genated blood. BV, body volume.
short ventral aorta. Blood flows through the gills into a stiff, long dorsal
482 INTEGRATION OF PHYSIOLOGICAL SYSTEMS
...............................................................................
gills and the air-breathing organ. In the tropical freshwa- suggested for the spongy heart of amphibians. In addition,
ter teleost Hoplerythrinus, the posterior gill arches give rise the absence of sinoatrial valves in the Channa heart and the
to the coeliac artery, which perfuses the swimbladder and arrangement of the veins probably play an important role
connects to the dorsal aorta by a narrow ductus. When the in preventing mixing of oxygenated and deoxygenated
animal is breathing water, most of the cardiac output is di- blood as these flows return to the heart in common vessels.
rected to the first two gill arches and flows to the body. Fol- Finally, muscular ridges on the wall of bulbus may prevent
lowing intake of air, the proportion of the blood flow to the mixing of the oxygenated and deoxygenated flows when
posterior gill arches and therefore to the swimbladder in- they are ejected from the heart. Once again this situation
creases, providing increased opportunity for oxygen uptake is similar to that seen in amphibians.
from the swimbladder. The division of the heart is more complete in the lung-
fishes (Dipnoi),which possess gills, lungs, and a pulmonary
circulation. The African lungfish, Protol)terus, has a partial
There are many more species of alr-breathing septum In the atrium and ventr~clcand 5piral folds in the
flsh in tropical than in temperate reglons. Why? bulh~lsiordis (Figure 12-17).This arrangement mantains
the srpdrdtlon of oxygenated ~ n deoxygenated
d blood in
the heart. The anterior gill arches lack lamellae and oxy-
genated blood can flow from the left side of the heart di-
The air-breathing fish Channa argus uses several mech- rectly to the tissues. Within the lamellae present in the pos-
anisms for achieving some separation of oxygenated and terior gill arches is a basal arterio-arterial connection that
deoxygenated blood in the circulation. The most important allows blood to bypass the lamellae when only the lung is
mechanism is a division of the ventral aorta into two ves- in operation (e.g., during estivation, a state of torpor oc-
sels, a posterior and an anterior ventral aorta. The anterior curring in the summer). Blood from the posterior gill arches
vessel supplies the first two gill arches and the air-breathing flows to the lungs or enters the dorsal aorta via a ductus.
organ, whereas the posterior vessel supplies the posterior The ductus is richly innervated and is undoubtedly involved
arches (Figure 12-16).The posterior arches are reduced in in controlling blood flow between the pulmonary artery
size, and the fourth arch is modifikd so that the afferent and and the systemic circulation. The initial segment of the pul-
efferent branchial arteries are in direct connection. Oxy- monary artery is muscular and is referred to as the pul-
genated blood is preferentially directed to the posterior monary vasomotor segment. This vasomotor segment and
arches, and the deoxygenated blood to the first two arches. the ductus probably act in a reciprocal fashion: when one
This is achieved without division of the heart. The ventri- constricts, the other dilates. The ductus in lungfish is anal-
cle, however, is spongy (trabeculate), which may serve to ogous to the ductus arteriosus of fetal mammals, acting as
prevent the mixing of blood in the ventricle, as has been a lung bypass when the lung is not functioning.
Efferent
branchial
artery
Afferent
branchial
artery
~ulbus Ventricle
Figure 12-16 Even though the heart of the air-breathingteleost Channa dorsal aorta. The fourth gill arch is modified so that the afferent and ef-
argus is undivided, the flows of oxygenated and deoxygenated blood are ferent branchial arteries are connected. Compare with Figure 12-15,
partially separated. Deoxygenated blood (black arrows) preferentially which illustrates the circulation of more typical water-breathing teleosts.
flows through the first two gill arches and a~r-breathingorgan, whereas [Adaptedfrom lshirnatzu and Itazawa, 1993.1 .
oxygenated blood (red arrows) flows through posterior arches into the
Tissues
Dorsal aorta A
~ u l b u scordis
' ~enkicle~ t r i u m EJOxygenated blood
Figure 12-17 The circulation of the African lungfish, Protopterus, is tinct pulmonary circulation. The absence of lamellae in the anterior gill
marked by nearly complete separation of oxygenated blood (red arches permits blood to flow directly to the systemic circulation via the
arrows) and deoxygenated blood (black arrows). This separation is dorsal aorta. The ductus and pulmonary vasomotor segment act recip-
achieved by a septum dividing the atrial and ventricular chambers and a rocally to direct blood to the dorsal aorta or lungs depending on whether
long spiral fold in the bulbus cordis. This fish possesses a lung and dis- the fish is breathing in water or air. [Adapted from Randall, 1994.1
Sub-
lrnocutaneous
Amphibians
Amphibia have two completely separated atria, but a sin-
gle ventricle. In the frog heart, the oxygenated and deoxy-
genated blood is separated even though the ventricle is un-
divided. Oxygenated blood from the lungs and skin is
preferentially directed toward the body via the systemic
arch, whereas deoxygenated blood from the body is di-
rected toward the pulmocutaneous arch. This separation of
oxygenated and deoxygenated blood is aided by a spiral
fold within the conus arteriosus of the heart (Figure 12-18).
Deoxygenated blood leaves the ventricle first during systole
and enters the lung circulation. Pressures rise in the pul-
mocutaneous arch and become similar to those in the sys-
temic arch. Blood then begins flowing into both arches,
with the spiral fold partially dividing the systemic and pul-
mocutaneous flows within the conus arteriosus.
The volume of blood going to the lungs or body is in-
- -
'~ertlcal septum
Horizontal
septum
Y Cavum venosum
-
Cavum venosum
Common
pulmonary
artery
Cavum venosum
and cavum
pulmonale
..
I I I I I I I I I I I I I I I I I I
Time (s) Time (100 ms)
Figure 12-20 In turtles, the pressures in the systemic and pulmonary out- at the indicated sites during a single heartbeat in (A) a turtle, Chrysemys
flows are nearly identical during systole, whereas in veranid lizards they scripta, and (B) a monitor lizard, Varanus exanthematicus. [Part Afrom
differ considerably. Shown are blood pressures measuredsimultaneously Shelton and Burggren, 1976; part B from Burggren and Johansen, 1982.1
tricle; consequently, the valves at the base of the left sys- of systole. Under some experimental circumstances peak
temic arch remain closed throughout the cardiac cycle right ventricular pressure becomes equal to left ventricular
(Figure 12-22C).All blood ejected from the right ventricle pressure and exceeds left systemic pressure. As a result, the
passes into the pulmonary artery and flows to the lungs. valves at the base of the left systemic arch open, and blood
Thus, the crocodilian reptile is functionally the same as the from the right ventricle is ejected into the systemic circula-
mammal in that there is complete separation of systemic tion during late systole (Figure 12-22D,E). In this case, a
portion of the deoxygenated blood returning to the heart
-
and pulmonary blood flow.
Crocodilian reptiles, however, have the added capacity from the body via the right atrium is recirculated in the sys-
This P -
to shunt blood from the pulmonary to the systemic circuit.
S shunt is achieved by active closure of a valve
at the base of the pulmonary outflow tract towards the end
temic circuit. Exactly when the P S shunt operates
normally in the animal is not clear. The role of the foramen
of Panizzae also remains enigmatic; it is only open during
A Diastole B Systole
Left
atrium
Atrioventricular
Figure 12-21 In veranid lizards, a pressure-tightseparation between the against the outer heart wall, forming a pressure-tight barrier. Deoxy-
cavum pulmonale and cavum venosum occurs during systole. (A) During genated blood remaining in the cavum venosum from the preceding di-
diastole, the muscular ridge only partially separates the cavum venosum astole is mixed with oxygenated blood from the cavum arteriosum and
and cavum pulmonale. Thus, oxygenated blood (red arrows) remaining flushed into the aortic arches. Deoxygenated blood with an admixture of
in the cavum venosum from the preceding systole is washed into the oxygenated blood is expelled from the cavum pulmonale into the pul-
cavum pulmonale by deoxygenated blood (black arrows). The cavum ar- monary arch. With no connection between the cavum venosum and
teriosum is filled with oxygenated blood. Separation between the cavum cavum pulmonale, different pressures can develop in the outflow tracts.
arteriosum and the cavum venosum is provided by at least one atrioven- [Adapted from Heisler et al., 1983.1
tricular valve. (B) During systole, the muscular ridge is pressed tight
486 I N T E G R A T I O N O F P H Y S I O L O G I C A L SYSTEMS
. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. . .
A Diastole
P -
Figure 12-22 Under some condltlons, a
S shunt operates durlng late systole in
crocodiles These schemat~cdlagrams and
pressure and flowtraclngs illustrate what hap-
oramen of panizza pens durlng the cardlac cycle with and wtthout
the shunt See text for d~scusslon[Adapted
from Jones, 1995 ]
c No P+ S shunt E P+ Sshunt
diastole, allowing flow between the aortic arches as the ing development to form the multi-chambered heart of the
heart relaxes. postnatal animal. The right side pumps blood to the lung,
the left side pumps blood around the body. Blood returning
Mammals and birds from the lungs enters the left atrium, passes into the left ven-
The heart in both mammals and birds, which consists of tricle, and is then ejected into the body circulation. Blood
four chambers, is in fact two hearts beating as one. The from the body collects in the right atrium, passes into the
heart originates as two separate tubes that join together dur- right ventricle, and is pumped to the lungs (seeFigure 12-3).
Valves prevent backflow of blood from the aorta to the Most of the blood ejected by the right ventricle is re-
ventricle, the atrium, and the veins. These valves are pas- turned to the systemic circuit via the ductus arteriosus.
sive and are opened and closed by pressure differences
Blood flow through the pulmonary circulation is greatly
between the heart chambers. The atrioventricular valves
reduced.
(bicuspid and tricuspid valves) are connected to the ven-
tricular wall by fibrous strands (see Figure 12-4).These A marked right-to-left (P +S) shunt operates; that
strands prevent the valves from being everted into the atria is, blood flows from the pulmonary to the systemic
when the ventricles contract and intraventricular pressures circuit.
are much higher than those in the atria. The walls of the
ventricle, especially the left chamber, are thick and muscu- At birth, the lungs are inflated, reducing the resistance
lar.'The inner surface of the ventricular muscle, or my- to flow in the pulmonary circuit. Blood ejected from the
ocardium, is lined by an endothelial membrane, the endo- right ventricle passes into the pulmonary vessels, resulting
cardium. The ventricular myocardium is covered by the in an increased venous return to the left side of the heart. At
epicardium. the same time, the placental circulation disappears, and the
resistance to flow increases markedly in the systemic circuit.
Mammalian fetus At birth, mammals shift from a placen- Pressures in the systemic circuit rise above those in the pul-
tal to a pulmonary circulation, a process that involves sev- monary circulation; if the ductus arteriosus fails to close af-
eral central cardiovascular readjustments. The lungs of the
mammalian fetus are collapsed, presenting a high resistance
to blood flow. In the fetus, the pulmonary artery is joined
-
ter birth, this pressure difference results in a left-to-right
(S P) shunt with blood flowing from the systemic to
the pulmonary circuit. Generally, however, the ductus arter-
to the systemic arch via a short, but large-diameter, blood iosus becomes occluded, and blood flow through the duc-
vessel, the ductus arteriosus (Figure 12-23).Heart function tus does not persist.
in the mammalian fetus exhibits three important features: If the ductus arteriosus remains open after birth, blood
flow to the lungs exceeds systemic flow, because a portion
of the left ventricular output passes via the ductus into the
pulmonary artery and to the lung. In these circumstances,
systemic flow is often normal, but pulmonary flow may be
twice the systemic flow, and cardiac output from the left
ventricle may be twice that from the right ventricle. The re-
sult is a marked hypertrophy of the left ventricle. The work
done by the left ventricle during exercise is also much
greater than normal, and the capacity to increase output is
limited. As a result, the maximum level of exercise is much
reduced if the ductus arteriosus remains open after birth.
Furthermore, this condition increases the blood pressure in
the lungs, leading to a greater fluid loss across lung capil-
lary walls and to possible pulmonary congestion. These
problems only become detrimental when the left ventricle
has become enlarged. An open ductus arteriosus is readily
and easily correctable by surgery.
Fetal blood is oxygenated in the placenta and mixed
with the blood returning from the lower body via the in-
ferior vena cava, a vein that in turn empties into the right
atrium (see Figure 12-23). In the interatrial septum is a
hole, the foramen ovale, that is covered by a flap valve;
oxygenated blood returning via the inferior vena cava
is directed into the left atrium through the foramen ovale.
Oxygenated blood is then pumped from the left atrium
into the left ventricle and ejected into the aorta, whence it
flows to the head and upper limbs. Deoxygenated blood
Figure 12-23 In the mammalian fetal heart, most of the blood ejected returning to the right atrium via the superior vena cava
from the right ventricle returns to the systemic circulation via the duaus is preferentially directed toward the right ventricle,
arteriosus. Oxygenated blood returning from the placenta is shunted whence it flows into the systemic circuit via the ductus ar-
from the right to the left atrium through the foramen ovale and then
teriosus. At birth, pressures in the left atrium exceed the
pumped into the aorta. After birth, the ductus arteriosus normallycloses,
so systemic and pulmonary circulations become separated. The numbers
pressure in the right atrium; as a result, the foramen ovale
refer to the percentage of the combined cardiac output from the right closes, but its position is later indicated by a permanent
and left ventricles that flows to and from different regions of the body. depression.
488 INTEGRATION OF PHYSIOLOGICAL SYSTEMS
.......................................
Bird embryo A network of blood vessels, forming the Capillaries
8 8
chorioallantois, lies just under the shell of bird eggs. Oxy- Arterioles I Venules
gen diffusing across the eggshell is taken up by blood pass- 5000 r I I I I I I
HEMODYNAMICS
As we have noted, contractions of the heart generate blood
flow through the vessels-arteries, capillaries, and veins-
that form the circulatory system. Before examining the
properties of these vessels in detail, it is necessary to discuss
the general patterns of blood flow in these vessels and the re-
lationship between pressure and flow in the circulatory sys-
tem. The laws describing the relationships between pressure
and flow apply to both open and closed circulatory systems.
In vertebrates and other animals with a closed circula-
tion, the blood flows in a continuous circuit. Since fluids are Figure 12-24 Blood velocity is inversely proportional to the cross-
incompressible, blood pumped by the heart must cause sectional area of the circulation at any given point. Blood velocity is high-
flow of an equivalent volume in every other part of the cir- est in the arteries and veins and lowest in the cap~llaries;the converse is
culation. That is, at any one time the same number of liters true for the cross-sectional area. [Adapted from Feigl, 1974.1
Laminar and Turbulent Flow Figure 12-25 Blood flow through smaller vessels approximates contin-
In many smaller vessels of the circulation, blood flow uous laminarflow, but in large elastic arteries pulsatile larninarflow is ob-
served. As shown in these velocity profiles,the flow rate is higher towards
is streamlined. Such continuous laminar flow is character- the center of the vessel. (A) The presence of red blood cells flattens the
ized by a parabolic velocity profile across the vessel profile of blood compared with that of plasma. (6) Pulsatile flow is marked
(Figure 12-25A). Flow is zero at the wall and maximal at by a flat profile and reversal of flow during each heartbeat.
CIRCULATION 489
......................................
the center along the axis of the vessel. A thin layer of blood strenuous exercise. The highest flow rates in the mam-
adjacent to the vessel wall does not move, but the next layer malian circulation are in the proximal portions of the
of fluid slides over this layer, and so on, each successive aorta and pulmonary artery, and turbulence may occur
layer moving at an increasingly higher velocity, with the distal to the aortic and pulmonary valves at the peak of
maximum at the center of the vessel. A pressure difference ventricular ejection or during backflow of blood as these
supplies the force required to slide adjacent layers past each valves close. In general, flow will be turbulent in portions
other, and viscosity is a measure of the resistance to sliding of the circulation where vessel walls are smooth and
between adjacent layers of fluid. An increase in viscosity the vessels are undivided only if Re is greater than about
will require a larger pressure difference to maintain the 1000, a value seldom observed. Small back eddies may
same rate of flow, as discussed later. form at arterial branches and, like the back eddies in rivers,
The pulsatile laminar flow characteristic of large arter- can become detached from the main flow regime, being
ies has a more complex velocity profile than the continuous carried downstream as small discrete regions of turbu-
laminar flow characteristic of smaller vessels. In large ar- lence. These eddies can form in the circulation when the Re
teries, blood is first accelerated and then slowed with each is as low as 200.
heartbeat; in addition, since the vessel walls are elastic, they
expand and then relax as pressure oscillates with each Relationship between Pressure and Flow
heartbeat. Close to the heart, the direction of flow reverses Flow will occur between two sites where there is a differ-
each time the aortic valves shut. The end result is that the ence in potential energy, which can be measured as a dif-
velocity across large arteries has a much flatter profile than ference in pressure. Thus differences in pressure between
the velocity across more peripheral blood vessels and the di- two points in a flow path establish a pressure gradient and
rection of flow oscillates (Figure 12-25B). therefore the direction of flow- from high to low pressure.
In turbulent flow fluid moves in directions not aligned (An exception is a fluid at rest under gravity, where pressure
with the axis of the flow, thus increasing the energy needed increases uniformly with depth but flow does not occur.) ,
to move fluid through a vessel. Laminar flow is silent; tur- When the heart contracts, the potential energy (pressure)in
bulent flow noisy. In the bloodstream, turbulence causes vi- the ventricle increases. Pressures generated by heart con-
brations that produce the sounds of the circulation. Detec- tractions are dissipated by the flow of blood, because en-
tion of these sounds with a stethoscope can localize the ergy is used to overcome the resistance to flow through the
points of turbulence. Blood pressure measurement with a vessels. For this reason, blood pressure falls as the blood
sphygmomanometer depends on hearing the sounds asso- passes from the arterial to the venous side of the circulation
ciated with blood escaping past the pressure cuff as blood (Figure 12-26).
pressure rises during systole. Sounds can be heard in vessels
when blood velocity exceeds a certain critical value and in
Capillaries
heart valves when they open and close. I I
Although turbulent flow is uncommon in the peripheral Arterioles! ! Venules
circulation, it does occur in some situations. The Reynolds
number (Re) is an empirically derived value that indicates
whether flow will be laminar or turbulent under a particu-
lar set of conditions. A high Reynolds number indicates
flow will be turbulent, whereas a low number indicates
flow will be laminar. The Re is directly proportional to the
flow rate, Q (in milliliters per second), and density, p, of the
blood, and inversely proportional to the inside radius of the
vessel, r (in centimeters), and viscosity, q, of the blood:
I Large veins I $s
Tunics
adventitia ,Tunica adventitia
elastic
p&q membrane
membrane
Figure 12-27 In the mammalIan per~pheralc~rculatlon,blood flows from larger vessels, the endothel~umIS surrounded by a muscle layer (tunlca
the heart v~aprogresslvely smaller arteries, then through the mlcroclrcu- medla) and outer flbrous layer (tun~caadvent~t~a)
[Adapted from M a r t ~ n ~
lat~on,and finally back to the heart v ~ aprogresslvely larger velns A layer and T~mmons,1995 ]
of endothellal cells, the endothellurn, llnes the lumen of all vessels In
is ejected into the arterial system, pressure rises and the According to Laplace's law, the wall tension required to
vessels expand. As the heart relaxes, blood flow to the pe- maintain a given transmural pressure within a hollow
riphery is maintained by the elastic recoil of the vessel structure increases with increasing radius (see equation
walls, resulting in a reduction in arterial volume (see 12-1).Elastic vessels thus are unstable and tend to balloon;
Figure 12-28). If arteries were simply rigid tubes, pressures that is, since they cannot develop high wall tension as pres-
and flow in the periphery would exhibit the same stops sure increases, they tend to bulge out. In blood vessels, this
and starts that occur at the exit of the ventricle during each instability is prevented by a collagen sheath that limits their
heartbeat. Although elastic, arteries become progressively expansion. Ballooning of a blood vessel (aneurism) can oc-
stiffer with increasing distension. As a result, they are cur, however, if the collagen sheath breaks down.
easily distended at low pressures, but then resist further In general, the elasticity of the arterial wall, as well as
expansion at high pressures. The response of arterial walls the thickness of the muscular layer, decreases with increas-
to distension is similar in a wide variety of animals, re- ing distance from the heart. That is, farther from the heart,
flecting similar structural and functional characteristics the arteries become more rigid and serve primarily as blood
(Figure 12-29). conduits. For example, the aorta of a dog becomes
INTEGRATION OF PHYSIOLOGICAL SYSTEMS
.................................................................
Arterial
oscillations
Ventricular
/ pressure
I oscillations Damping
\
oscillations
Figure 12-28 The systemic arterial system functions as a conduit and sistance of arteries account for the pressure-reservoirfunction (2) and
pressure reservoir; it also smoothes out pressure oscillations and controls damping of oscillations in pressure and flow (3). Controlled hydraulic re-
distribution of flow to capillaries. The conduit function (1) is served by the sistance in the peripheral vascular beds controls the distribution of blood
vascular channels along which blood flows toward the peripheryw~thmin- t o the various tissues (4). [Adapted from Rushmer, 1965a.l
imal frictional loss of pressure.The distensible walls and high outflow re-
progressively stiffer and its diameter decreases with in- tic arch narrows rapidly and becomes much more rigid
creasing distance from the heart (Figure 12-30).In a whale, than that of a dog. The elastic whale aortic arch expands
the aortic arch at the exit from the heart is very elastic and with each heartbeat, accommodating about 50% -75% of
has a large diameter, but the arterial system beyond the aor- the stroke volume; the remainder flows into the portion of
the arterial system downstream of the aortic arch. The
change in ventricular volume with each heartbeat can be as
much as 35 liters in a large whale, with a heart rate of
around 12- 18 per minute.
The extent of elastic tissue in arteries varies depending
on the particular function of each vessel. In fishes, for ex-
ample, blood pumped by the heart is forced into an elastic
bulbus and a ventral aorta (see Figure 12-15).The blood
then flows through the gills and passes into a dorsal aorta,
the main conduit for the distribution of blood to the rest of
the body. A smooth, continuous flow of blood is required
in the gill capillaries for efficient gas transfer. The bulbus,
the ventral aorta, and the afferent branchial arteries lead-
ing to the gills are very compliant and act to smooth and
maintain flow in the gills in the face of large oscillations
produced by contractions of the heart. The dorsal aorta,
which receives blood from the gills, is much less elastic than
the ventral aorta. If the dorsal aorta were more elastic than
the ventral aorta, there would be a rapid rush of blood
Relative distension (PIP,) through the gills during each heartbeat. This rush would in-
crease rather than decrease the oscillations in flow through
Figure 12-29 The elastic properties of arteries are surprisinglysimilar in
the gills. In this example, then, to ensure a steady blood
a wide variety of animals, with nautilus and octopus being notable ex-
ceptions. This similarity is reflected in plots of elastic modulus versus rel- flow through the gill capillaries, the major compliance must
ative distension, expressed as pressure (P) divided by the resting blood be placed before, not after, the gills to dampen the oscil-
pressure (P,) of the species. [Adapted from Shadwick, 1992.1 lations in flow through the gills. The ventral aorta must
CIRCULATION 495
......................................
Ventral aortic
blood flow
(ml min-') 30
Time
-
I
m
near the heart. Aortic blood pressures greater than 195
mm Hg have been recorded in an anesthetized giraffe
whose head was raised about 1.5 meters (Figure 12-33B).
-$
E
E
50
Arterial pressures in the legs of the giraffe are even higher
than aortic pressures; to prevent the pooling of blood, the
V)
giraffe has large quantities of connective tissue surround-
n
s? ing the leg vessels. As the giraffe lowers its head to the
ground, arterial blood pressure at the level of the heart is
reduced considerably, thus maintaining a relatively con-
stant blood flow to the brain. The wide variation in aor-
tic pressure as the giraffe moves its head position could
Time (ms) lead to extensive pooling of blood (head raised) or de-
Figure 12-32 In the aorta of mammals and birds, the peak blood pres-
creased flow (head lowered) in arterioles other than those
sure and pressure pulse both increase with distance from the heart. of the head. Pooling most likely is prevented by vasocon-
Shown are simultaneous recordings of a rabbit's blood pressure in the striction of these peripheral vessels when the head is raised.
aortic arch (2 cm from the heart) and at trifurcation of the aorta (24 crn Conversely, when the head is lowered, extensive vasodila-
from the heart). Note that the mean pressure is slightly less attrifurcation
tion of arterioles leading to capillary beds other than those
of the aorta than in the aortic arch near the heart. [Adapted from Langille,
1975.)
-
ies in the head, chest, and limbs. Once a person moves to
a sitting or standing position, the relationship between the
head, heart, and limbs changes with respect to gravity, and Vasoconstriction Vasodilation of
the heart is now a meter above the lower limbs. The result of vessels in Aortic pressure vessels in
is an increase in arterial pressure in the lower limbs and a lower body decreases lower body
decrease in arterial pressure in the head. The height of the
column of blood simply results in a higher blood pressure
due to gravity.
Gravity has little effect on capillary flow, which is de-
termined by the arterial-venous pressure difference rather
than the absolute pressure. That is, gravity raises arterial
and venous pressure by the same amount and therefore
does not greatly affect the pressure gradient across a capil-
lary bed. Because the vascular system is elastic, however, an
increase in absolute pressure expands blood vessels, partic-
ularly the compliant veins. Thus, pooling of blood tends to
occur, particularly in veins, in different regions of the body
as an animal changes position with respect to gravity. This I I I I I I
effect is related solely to the elasticity of blood vessels and 0 10 20 30 40 50 60
would not occur if the blood flowed in rigid tubes. Time (s)
The problems of pooling and maintaining capillary
Figure 12-33 As animals with long necks raise and lower their heads, the
flow are acute in species with long necks. For instance, cardiovascular system must adjust to maintain blood flow to the brain and
when the giraffe is standing with its head raised, its brain avoid pooling of blood in the lower part of the body. See text for discus-
is about 6 meters above the ground and over 2 meters sion. [Adapted from White, 1972.1
CIRCULATION 497
........................................
in the head probably maintains flow despite the lower aor- tices in the blood ejected into the aorta during systole. In
tic pressure. most other parts of the circulation, flow is laminar, and os-
The ability of the giraffe to regulate pressure and flow cillations in velocity are damped by the compliance of the
in peripheral vessels other than those to the head is partic- aorta and proximal arteries.
ularly crucial for kidney function. If the kidney tubule were Mean velocity in the aorta-the point of maximal
subjected to the enormous changes in blood pressure asso- blood velocity-is calculated as about 33 cm -s-' in hu-
ciated with the raising and lowering of the giraffe's head, mans, based on a cross-sectional area of about 2.5 cm2and
the rate of glomerular filtration would be chaotic. Each cardiac output of about 5 La min-'. If we assume that max-
time the animal lifted its head, the large increase in arterial imal velocity in a vessel is twice the mean velocity (valid
blood pressure would lead to a very high rate of ultrafiltrate only if the velocity profile is a parabola), then the maximal
formation in the kidney; this in turn would require that velocity of blood flow in the human aorta would be 66
fluid be reabsorbed at an equally high.rate. In the absence cm-s-l. If cardiac output is increased by a factor
of any appropriate controls, the giraffe could lower its head of 6 during heavy exercise, maximal velocity is raised
to drink and then lose any fluid gained as it was filtered to 3.96 m-s-l. In contrast, the pressure pulse associated
through the kidney when the head was raised. Thus, the gi- with each heartbeat travels through the circulation at
raffe must have mechanisms for adjusting peripheral resis- -
3-35 m s-l; thus, the pressure pulse travels faster than the
tance to flow in various capillary beds as it swings its head flow pulse.
from ground level to drink to a height of 6 m to eat. Simi-
lar problems must have been or are faced by a number of Venous System
other animals with long necks, like dinosaurs and camels. The venous system acts as a conduit for the return of blood
Pooling of blood, with changes of position with respect from the capillaries to the heart. It is a large-volume, low-
to gravity, is not a problem for animals in water, because pressure system consisting of vessels with a larger inside di-
the density of water is only slightly less than that of blood, ameter than the corresponding arteries (see Figure 12-27).
whereas air is much less dense than blood. In water, the hy- In mammals, about 50% of the total blood volume is con-
drostatic pressure increases with depth, and effectively tained in veins (see Figure 12-3). Venous pressures seldom
matches the increase in blood pressure due to gravity; thus exceed 11rnrn Hg (1.5 kPa), roughly 10% of arterial pres-
transmural pressure does not change, so the blood does not sures. The walls of veins are much thinner, contain less
pool. Clearly, the circulatory problems faced by tall terres- smooth muscle, and are less elastic than arterial walls; ve-
trial dinosaurs were very different from those of aquatic nous walls also contain more collagen than elastic fibers. As
dinosaurs. a result, the walls of veins are easily stretched and exhibit
much less recoil than occurs in arteries. The large diame-
Velocity of arterial blood flow ter and low pressure of veins permits the venous system to
Blood flow and the oscillations in flow with each heartbeat function as a storage reservoir for blood. If venous pres-
are greatest at the exit to the ventricle, decreasing with in- sures were high, then according to Laplace's law (seeequa-
creasing distance from the heart (Figure 12-34).At the base tion 12-l),very high wall tensions would develop, requir-
of the aorta, as noted earlier, flow is turbulent and reverses ing the walls to be very strong to prevent them from
during diastole as closure of the aortic valves creates vor- tearing.
In the event of blood loss, venous blood volume, not ar-
terial volume, decreases in order to maintain arterial pres-
sure and capillary blood flow. The decrease in the venous
blood reservoir is compensated for by a reduction in venous
volume. The walls of many veins are covered by smooth
muscle innervated by sympathetic adrenergic fibers. Stimu-
lation of these nerves cause vasoconstriction and a reduc-
tion in the size of the venous reservoir. This reflex allows
some bleeding to take place without a drop in venous
pressure. Blood donors actually lose part of their venous
reservoir; the loss is temporary, however, and the venous
system gradually expands as blood is replaced due to fluid
retention.
,\ \ I'
\\ Pericyte covered with ;
Figure 12-36 A m~croc~rculatory bed conslsts of small arter~es(arterl- from the arter~alto venous system can occur vla the thoroughfare chan-
oles), cap~llar~es,
and venules Cap~llar~es,
wh~chconslst of a s~nglelayer nel, but most blood flows through the network of cap~llar~es The pre-
of endothel~alcells surrounded by a basement membrane and have oc- cap~llarysphlncterhelps regulate flow Into the capillary bed Also see Fig-
cas~ondcontractile perlcyte cells wrapped around them. D~rectflow ures 12-27 and 12-37
500 INTEGRATION OF PHYSIOLOGICAL SYSTEMS
...............................................................................
muscle that becomes discontinuous in the metarterioles and A Continuous capillary
ends in a smooth muscle ring, the precapillary sphincter.
Capillary walls, which are completely devoid of connective
tissue and smooth muscle, consist of a single layer of en-
dothelial cells surrounded by a basement membrane of col-
lagen and mucopolysaccharides. The capillaries are often
categorized as arterial, middle, or venous capillaries, the
latter being a little wider than the other two types. A few
elongated cells with the ability to contract, called pericyte
cells, are found wrapped around capillaries. The venous
capillaries empty into pericytic venules, which in turn join (mucopolysaccharides
and collagen fibers)
the muscular venules and veins. The venules and veins are
valved, and the muscle sheath appears after the first post-
capillary valve. Even though the walls of capillaries are thin B Fenestrated capillary
and fragile, they require only a small wall tension to resist
stretch in response to pressure because of their small diam-
eter (see equation 12-1).
The innervated smooth muscle of the arterioles and, in
particular, the smooth muscle sphincter at the junction of
arteries and arterioles control blood distribution to each
capillary bed. Most arterioles are innervated by the sym-
pathetic nervous system; a few arterioles (e.g., those in the
lungs) are innervated by the parasympathetic nervous sys-
tem. Different tissues have varying numbers of capillaries
open to flow and show some variation in the control of
~ a g e m e nmembrane
t
blood flow through the capillary bed. In some tissues, open-
ing and closing of the precapillary sphincters, which are not
innervated and appear to be under local control, alter c Sinusoidal capillary
blood distribution within the capillary bed. In other tissues,
however, most, if not all, of the capillaries tend to be open
(e.g., in the brain) or closed (e.g., in the skin) for consider-
able periods. All capillaries combined have a potential vol-
ume of about 14% of an animal's total blood volume. At
any one moment, however, only 30%-50% of all capil-
laries are open, and thus only 5% -7% of the total blood
volume is contained in the capillaries.
Small
artery l5 r
-
1s 1 min
C--l
(500 pm) 5
L
Metarteriole
(70 ~ m )
a,
15 [-
0
Arteriole f'15
caiary [d
(25 w) ;
- 0
L Figure 12-38 The pressure pulse is reduced and the mean blood pres-
sure decreases as blood flows through a cap~llarybed. (A) Blood pressure
capillary g tracings recorded throughout capillary bed of the frog mesentery. The
pressure is smoothed and falls as blood flows across the capillary bed.
(B) Plot of blood pressure versus location in circulation of the subcuta-
- -
1 min
neous layers of the bat wing. Shaded area represents 2 1 SE (standarder-
ror) from the average values indicated by thick line. Also plotted are typ-
ical tissue and lymphatic pressures for comparison. [Part A from
, Weiderhielm et al., 1964; part B from Weiderhielm and Weston, 1973.1
capillary. The fluid, however, does not accumulate ln the tis- Capillary
sues, but 1s drained by the lymphatic system and returned
to the circulation. Thus, there typically is a circulation of end
fluid from the arterial end of capillary bed into the mtersti-
tial spaces and back into the blood across the venous end
Filtration Uptake
of the capillary bed or via the lymphatic system. Because of
this bulk flow of fluid, the exchange of gases, nutrients, and
wastes between blood and tissues exceeds that expected by
diffusion alone.
Net filtration of fluid across capillary walls will result
in an increase in tissue volume, termed edema, unless the 9
3
excess fluid is carried away by the lymphatic system. In the m
(0
E
kidney, capillary pressure is high and filtration pressures ex- a
ceed collold osmotlc pressures; hence, an ultrafiltrate is
formed in the kidney tubule, eventually to form urine. The osmotic pressure
kidney is encapsulated to prevent swelling of the tissue in
the face of ultrafiltratlon. In most other tissues, there is only 1
a small net movement of fluid across capillary walls and tis- Arterial Venous
end end
sue volume remains constant. A rise in capillary pressure,
owing to a rise in either arterial or venous pressure, will re- Capillary length
sult in increased loss of fluid from the blood and tissue Figure 12-39 Net fluid flow across the capillary wall depends on the dif-
edema. In general, though, arterial pressure remains fairly ference between the blood pressure and the colloid osmotic pressure of
constant t o prevent large oscillations in tissue volume. A the extracellular fluid. At the arterial end of the capillary, blood pressure
drop in colloid osmotic pressure can result from a loss of exceeds colloid osmotic pressure and fluid is filtered from the plasma into
the extracellular space (area 1). At the venous end, the reverse is true and
protein from the plasma by starvation or excretion or by in-
fluid is drawn back into the plasma from the extracellular space (area 2).
creased capillary wall permeability, leading to movement of Area 1 is somewhat larger than area 2 in most capillary beds; that is, there
plasma proteins into the interstitial space. If filtration pres- is a small net loss of fluid from the circulationto the extracellular space. In
sure remains constant, a decrease in colloid osmotic pres- general, this tissue fluid is drained and returned to the bloodstream via
sure will also result In an increase in net fluid loss to the tis- the lymphatic system.
sue spaces.
CIRCULATION 503
....................................
A Lyrnphatlc trunks
F 10 s 1 min
receptor
I Develops
into antibody
secreting
plasma cell .
Secretes
cytokines,
which promotes
growth and
responsiveness
of B and Tc
cells
IDevelops
into active CTL,
which destroys
altered self-cell
I Antibodies bind
to pathogen,
which are
eliminated by
phagocytes
Cytokines
ing the cell for degradation by phagocytes. T, cells can rec- a result of these and other interactions, the cells adhere to
ognize tumor cells and pathogen-infected tumor cells; upon the endothelium. Once firmly adhered, the leukocytes can
recognizing such cells, Tc are stimulated to mature into ac- move between the endothelial cells and migrate into the in-
tive cytotoxic T lymphocytes (CTLs),which destroy the al- fected tissue.
tered self-cells. Recognition
- of antigen by T, cells stimu-
A A
Sectional view of
vascular endothelium
/
Leukocyte migration
into infected tissue /
ICA
+
Selection tethering to LFA-1 Integrin-ICAM
inflamed endothelium production interaction
Figure 12-42 Leukocytes mlgrate from the clrculat~onInto tlssues at actlons between cell-surface molecules that cause leukocytes to adhere
sltes of lnflamrnatlon (A) Ovewlew of leukocyte adherence to and ex- to an Inflamed endothellurn [Adaptedfrom Kuby, 1997 I
travasatlon across Inflamedvascular endothellurn (B) Some of the Inter-
506 INTEGRATION OF PHYSIOLOGICAL SYSTEMS
...............................................................................
Control of the Central Cardiovascular System into two functional regions, which have opposing effects on
Baroreceptors monitor blood pressure at various sites in pressure:
the cardiovascular system. Information from barorecep-
Stimulation of the pressor center results in sympathetic
tors, along with that from chemoreceptors monitoring
activation and a rise in blood pressure.
the CO,, '. O,,
'. and DHof the blood. is transmitted to the
brain. ~~~~l~ or changes
in the composition
of the extracellular fluid of muscles activate afferent fibers
Stimulation of the depressor center results in parasym-
pathetic activation and a drop in blood pressure.
embedded in muscle tissue, and this in turn causes
changes in the cardiovascular system. In addition, inputs In general terms, various sensory inputs affect the balance
from cardiac mechanoreceptors and from a variety of between pressor and depressor activity: some activate the
thermoreceptors lead to reflex effects on the cardiovascu- pressor center and inhibit the depressor center; others have
lar system. the reverse effect. Thus, the various inputs that converge on
In mammals, the integration of these sensory inputs oc- the medullary cardiovascular center are modified and inte-
curs in a collection of brain neurons referred to as the grated. The result is an output that activates the pressor or
medullary cardiovascular center, located at the level of the the depressor center and produces cardiovascular changes
medulla oblongata and pons. The medullary cardiovascu- in response to changing requirements of the body or distur-
lar center also receives inputs from other regions of the bances to the cardiovascular system. Figure 12-43presents
brain, including the medullary respiratory center, hypo- an overview of this central circulatory control in mammals.
thalamus, amygdala nucleus, and cortex. The output from
the medullary cardiovascular center is fed into sympathetic Arterial baroreceptors
and parasympathetic autonomic motor neurons that in- Baroreceptors, which are widely distributed in the arterial
nervate the heart and the smooth muscle of arterioles and system of vertebrates, show increased rates of firing with in-
veins, as well as to other areas of the brain such as the creases in blood pressure. Unmyelinated baroreceptors
medullary respiratory center. have been localized in the central cardiovascular system of
Stimulation of sympathetic nerves increases the rate amphibia, reptiles, and mammals. These unmyelinated
and force of contraction of the heart and causes vasocon- baroreceptors only respond to pressures above normal, ini-
striction; the result is a marked increase in arterial blood tiating reflexes that reduce arterial blood pressure and thus
pressure and cardiac output. In general, the reverse effects protect the animal from damaging increases in blood pres-
follow stimulation of parasympathetic nerves, the end re- sure. Myelhated baroreceptors, which have been found
sult being a drop in arterial blood pressure and cardiac out- only in mammals, respond to blood pressures below nor-
put. The medullary cardiovascular center can be divided mal, thus protecting the animal from prolonged periods of
I
I DISTURBANCE
I Cardiac
I Pulmonary
I
HEART AND
card~ovascular BLOOD lntravascular
"set point" center EFFECTORS VESSELS pressures
..
I
i
I
I----------- BARORECEPTOR
Figure 12-43 The circulatory control system in mammals involves a num- arterial set point, this center sends signals via autonomic nerves to main-
ber of negative-feedback loops. Various receptors monitor changes in tain an appropriate arterial blood pressure. The arterial set point is altered
the state of the cardiovascular system, sending inputs to medullary car- by inputs from other areas of the brain, which are in turn influenced by a
diovascular center. After integrating these inputs and comparing with the variety of peripheral inputs (dashed lines). [Adapted from Korner, 1971.]
CIRCULATION 507
...............................................................................
reduced blood pressure. Baroreceptors in the mammalian
carotid sinus have been studied much more extensively
than those in the aortic arch or subclavian, common
carotid, and pulmonary arteries. In mammals there appear
to be only minor quantitative differences between the
baroreceptors of the carotid sinus and the aortic arch. Birds
have aortic arch baroreceptors.
The carotid sinus in mammals is a dilation of the inter-
nal carotid at its origin, where the walls are somewhat thin-
ner than in other portions of the artery. Buried in the thin
walls of the carotid sinus are finely branched nerve endings V
-
baroreceptors in the toad are plotted (A)
immediately after the pressure increase
and (B) 45 seconds later. Each numbered
black line represents one observation cor-
responding to the pressure increase
shown on the horizontalaxis. The rapid ini-
tial peak response is greater than the re-
I I I I I I I I sponse 45 seconds after the pressure ~ n -
20 40 60 80 20 40 60 80 crease. [Adaptedfrom Van Vilet and West,
Step increase in pressure (mm Hg) Step increase in pressure (mm Hg) 1994.1
508 INTEGRATION OF PHYSIOLOGICAL SYSTEMS
.....................................*...
TABLE 12-1 changes in arterial pressure resulting from peripheral vaso-
Reflex effects observed during changes in carotid sinus constriction. Not surprisingly, there are many interactions
pressure
between the control systems associated with both the res-
Carotid sinus pressure* piratory and the cardiovascular system. For example, the
Autonomic effector Increased Decreased discharge pattern from stretch receptors in the lungs has a
Cardiac vagus ++++ -
marked effect on the nature of the cardiovascular changes
Cardiac sympathetic - +++ caused by chemoreceptor stimulation. If the animal is
breathing normally, changes in gas levels of the blood will
Splanchnic b e d
cause one set of reflex changes; if, however, the animal is
Resistance vessels -- ++ not breathing, chemoreceptor stimulation results in quite a
Capacitance vessels -- ++ different series of cardiovascular changes, as we will see in
Renal b e d ==0 + the h e r dscussion on dving.
Muscle b e d
Resistance vessels -- Cardiac receptors
Capacitance vessels - Both mechanoreceptiveand chemoreceptiveafferent nerve
Skin endings are located in various regions of the heart. Infor-
Resistance vessels - ++ mation on the state of the heart collected by these receptors
Capacitance vessels ?O 0 is transmitted via the spinal cord to the medullary cardio-
Adrenal catecholamines ==O ++ vascular center and other regions of the brain. In addition,
Ant~diuretichormone ? ++ stimulation of some cardiac receptors causes hormone re-
lease either directly from the atria or from other endocrine
+
*A means increased autonomic effect; a -, decreased autonomic ef-
tissues within the body. Stimulation of cardiac receptors
fect; and 0, no autonom~ceffect.
evokes a series of reflex responses including changes in
Source: Korner, 1971
heart rate and cardiac contractility and, under extreme con-
ditions, the pain that can be associated with a heart attack.
discharge frequency, causing a reflex increase in both car-
diac output and peripheral resistance, which tends to in- Atrial receptors The atrial walls contain many mechanore-
crease arterial pressure. Thus the baroreceptor reflex of the ceptive afferent fibers, which are classified into three types.
carotid sinus is a negative feedback loop that tends to sta- Myelinated A-type and B-type afferent fibers are embedded
bilize arterial blood pressure at a particular set point. The in the atria. A-fiber afferents respond to heart-rate changes
set point may be altered by interaction with other receptor and appear to relay information on heart rate to central
inputs or may be reset centrally within the medullary cardiovascular control centers. B-fiber afferents respond to
cardiovascular center by inputs from other regions of the increases in the rate of filling and volume of the atria. In-
brain (see Figure 12-43). creases in venous volume result in an increase in venous
pressure, which in turn raises the atrial filling and conse-
Arterial chemoreceptors quently the discharge frequency of the B-fibers. This in-
Arterial chemoreceptors, which are located in the carotid creased activity is processed by the central cardiovascular
and aortic bodies, are particularly important in regulating centers leading to two major effects, one on the heart and
ventilation (see Chapter 13),but they also have some effect one on the kidney. Stimulation of atrial B-fibers leads to a
on the cardiovascular system. These chemoreceptors re- faster heart rate mediated via increased activity in the sym-
spond with an increase in discharge frequency to an in- pathetic outflow to the sinus node of the heart. Stimulation
crease in CO, or to decreases in 0, and pH of the blood of these afferent fibers also causes a marked increase in
perfusing the carotid and aortic bodies. An increase in dis- urine excretion (diuresis),probably mediated by a decrease
charge frequency results in peripheral vasoconstriction and in antidiuretic hormone (ADH)levels in the blood. Thus,
a slowing of the heart rate if the animal is not breathing there is a negative feedback loop for regulating blood vol-
(e.g., during submersion). Cardiac output is reduced while ume. An increase in blood volume raises venous pressure
birds and mammals dive; peripheral vasoconstriction then and atrial filling; this stimulates atrial B-fibers, leading to
ensures the maintenance of arterial blood pressure and inhibition of ADH release from the pituitary. The resulting
therefore brain blood flow in the face of this reduction in fall in blood ADH levels leads to diuresis and therefore a re-
cardiac output. duction in blood volume.
Peripheral vasoconstriction can cause a rise in arterial The third type of atrial mechanoreceptor comprises un-
pressure, which then evokes reflex slowing of the heart by myelinated C-type afferent fibers innervating the junction
stimulation of the systemic baroreceptors. Nevertheless, of the veins and atria. Stimulation of these C-fiber afferents
stimulation of arterial chemoreceptors results in a slowing receptors affects both heart rate and blood pressure. If
of the heartbeat even when arterial pressure is regulated at heart rate is low, distension of this region results in an in-
a constant level. Chemoreceptor stimulation thus has a di- crease in heart rate, whereas if heart rate is high, stimula-
rect effect on heart rate, as well as an indirect effect via tion results in a fall in heart rate. Stimulation of C-fibers
CIRCULATION 509
......................................
also causes a fall in blood pressure. Both myelinated and ferents at low levels causes peripheral vasodilation and a re-
unmyelinated sympathetic fibers innervate the atria. Atrial duction in heart rate. Increased stimulation of these fibers
contraction and atrial distension reflexly stimulate these causes stomach relaxation and, at even higher frequencies,
fibers, causing an increase in heart rate. results in vomiting.
The atrial wall also contains stretch-sensitivesecretory
cells that produce atrial natriuretic peptide (ANP). This Skeletal muscle afferent fibers
hormone, which is released into the blood upon stretch of Somewhat surprisingly, most nerves innervating skeletal
these cells, has several endocrine effects. As its name indi- muscle contain more afferent fibers than efferent fibers.
cates, ANP causes an increase in urine production and The afferent fibers can be subdivided into four broad
sodium excretion, thereby effectively reducing blood vol- groups. Groups I and I1 are sensory fibers from muscle
ume and therefore blood pressure. ANP inhibits release of spindles and Golgi tendon organs; these seem to play lit-
renin by the kidney and production of aldosterone by the tle or no role in the control of the cardiovascular system.
adrenal cortex. It thus diminishes the renin-angiotensin- In contrast, stimulation of group 111 fibers, which are
aldosterone system, which stimulates sodium resorption myelinated "free nerve endings," or group IV fibers, which
and an increase in blood volume (see Chapter 14).In ad- are unmyelinated sensory endings, appears to have car-
dition to these actions, ANP inhibits release of ADH and diovascular effects. These fibers are activated by either
acts directly on the kidney to increase water and sodium mechanical or chemical stimulation, with most fibers re-
excretion. ANP has been demonstrated to have a depres- sponding to only one modality. Mechanical stimulation
sor effect, reducing both cardiac output and blood pres- may be due to contraction, squeezing, or stretching of the
sure. In addition ANP antagonizes the pressor effect of muscle. The changes in the extracellular fluid associated
angiotensin. with muscle contraction also are thought to stimulate
Atrial natriuretic peptide belongs to a family of natri- chemoreceptive muscle afferent fibers and evoke cardio-
uretic peptides (A-, B-, C-, and V-type natriuretic peptide) vascular changes. Large changes in pH and osmotic pres-
sharing a common 17-amino-acid ring structure linked by sure raise the activity of group IV fibers, but it is not
a disulfide bridge. Since the initial investigations of ANP in clear if the pH or osmotic changes occurring in vivo are
the early 1980s, natriuretic peptides have been found in a adequate to mediate cardiovascular effects.
wide variety of tissues, including the central nervous sys- Electrical stimulation of muscle afferents can result in
tem. In many instances they may have an autocrine or either an increase or a decrease in arterial blood pressure,
paracrine function. For example, receptors for natriuretic depending on the fibers being stimulated or the frequency
hormone have been located in both the atria and ventricles of stimulation of a particular group of afferent nerves.
of the hearts of several vertebrates. Binding of locally re- At low frequencies, stimulation of some afferent fibers
leased natriuretic hormone to these receptors may reduce results in a fall in arterial blood pressure, whereas stimu-
contractility, indicating a paracrine function within the lation of the same fibers at high frequencies results in a rise
heart. in blood pressure. Electrical stimulation of afferent nerves
from muscles usually causes a change in heart rate in the
same direction as thk change in blood pressure; that is, if
blood pressure is elevated, so is heart rate, and vice versa.
In those instances where electrical stimulation of muscle
afferents causes an increase in heart rate and cardiac out-
put, there is also a change in the distribution of blood
in the body. Blood flow to the skin, kidney, gut, and inac-
tive muscle is reduced, thus augmenting flow to the active
muscles.
Ventricular receptors The endings of both myelinated and The cardiovascular response evoked by muscle con-
unmyelinated sensory afferent fibers are embedded in the traction has been shown to disappear following dorsal
ventricle. The myelinated fibers are mechanoreceptive and root section, so the response is presumably reflex in origin,
chemoreceptive, with separate endings for each modality. resulting from stimulation of afferent fibers in the muscle.
The mechanoreceptive endings are stimulated by interrup- The response varies depending on whether muscle contrac-
tion of coronary blood flow. The chemoreceptive endings tion is isometric (static exercise) or isotonic (dynamic ex-
are stimulated by substances like bradykinin. At low stim- ercise). Static exercise is associated with an increase in arte-
ulation levels, these fibers cause increased sympathetic out- rial blood pressure with little change in cardiac output,
flow and decreased vagal outflow to the heart, raising car- whereas dynamic exercise results in a large increase in
diac contractility as well as blood pressure. At higher cardiac output with little change in arterial blood pres-
stimulation levels, these fibers are necessary for the percep- sure. The sensory inputs from muscle afferent fibers are
tion of pain in the heart. Myelinated afferent fibers are processed in the central cardiovascular center, leading to
much less numerous than unmyelinated C-fiber afferent stimulation of the autonomic nerves innervating the heart
endings in the left ventricle. Stimulation of the C-fiber af- and vessels, the efferent arm of the reflex arcs.
510 INTEGRATION OF PHYSIOLOGICAL SYSTEMS
.........................................
Control of the Microcirculation resistance via a-adrenoreceptor stimulation. At lower lev-
Capillary blood flow adjusts to meet the demands of the tis- els of circulating epinephrine, however, P-adrenoreceptor
sues. If the requirements change suddenly, as in skeletal stimulation dominates, producing an overall vasodilation
muscle during exercise, then capillary flow also changes. If and a decrease in peripheral resistance. Even at levels of ep-
requirements for nutrients vary little with time, as in the inephrine that produce vasodilation, it causes a rise in ar-
brain, then capillary flow also varies little. The regulation terial blood pressure by stimulating p-adrenoreceptors in
of capillary flow can be divided into two main types, ner- the heart, causing a marked increase in cardiac output.
vous control and local control. The p-adrenoreceptors can be divided into two sub-
groups: PI-adrenoreceptors, which are stimulated by both
Nervous control of capillary blood flow circulating catecholamines (epinephrine) and adrenergic
Nervous control serves to maintain arterial pressure by ad- nerve stimulation (norepinephrine) and 0,-adrenorecep-
justing resistance to blood flow in the peripheral circula- tors, which respond only to circulating catecholamines. In
tion. The vertebrate brain and heart must be perfused with the peripheral circulation, only p,-adrenoreceptors are
blood at all times. An interruption in the perfusion of the present, whereas PI-adrenoreceptors are found in the heart
human brain rapidly results in damage. Nervous control of and coronary circulation, where both circulating cate-
arterioles ensures that only a limited number of capillaries cholamines and neurally released norepinephrine can have
will be open at any moment, for if all capillaries were open, a marked effect.
there would be a rapid drop in arterial pressure and blood We can summarize these effects as follows:
flow to the brain would be reduced. The nervous control of
capillary flow operates under a priority system. If arterial Stimulation of sympathetic nerves generally causes pe-
pressure falls, blood flow to the gut, liver, and muscles is re- ripheral vasoconstriction and a rise in arterial blood
duced to maintain flow to the brain and heart. Most arte- pressure.
rioles are innervated by sympathetic nerves, which release
norepinephrine at their endings. Some arterioles, however, An increase in circulating catecholamines causes a de-
are innervated by parasympathetic nerves, which release crease in peripheral resistance, with a rise in arterial
acetylcholine at their endings, pressure because of concomitant stimulation of the
heart and a rise in cardiac output.
Sympathetic stimulation and circulating catecholamines
Binding of the catecholamine norepinephrine to a-adreno- The response in any vascular bed depends on several things:
receptors in the smooth muscle of arterioles usually causes the type of catecholamine, the nature of the receptors in-
vasoconstriction and therefore a decrease in diameter of the volved, and the relationship between stimulation of the
arterioles. This decrease in diameter causes an increase in receptors and the change in muscle tone. Although stimu-
resistance to flow, thus reducing blood flow through that lation of a-adrenoreceptors usually is associated with vaso-
capillary bed. The generalized effect of sympathetic stimu- constriction and that of p-adrenoreceptors with vasodi-
lation is peripheral vasoconstriction and subsequent rise in lation, this is not invariably the case. An additional
arterial blood pressure. This overall response is mediated complicating factor is that not all sympathetic fibers are
by binding of norepinephrine from nerve endings to adrenergic. In some instances, they may be cholinergic, re-
a-adrenoreceptors in vascular smooth muscle, resulting in leasing acetylcholine from their nerve endings. Stimulation
an increase in smooth-muscle tension. of sympatheticcholinergic nerves causes vasodilation in the
Stimulation of p-adrenoreceptors in arterial smooth vasculature of skeletal muscle.
muscle, however, often results in relaxation of the muscle The action of catecholamines is extensively modulated
and an increase in diameter of the arterioles (i.e., vasodila- by a variety of substances, including neuropeptide Y and
tion), thereby decreasing the resistance to flow and in- adenosine. Neuropeptide Y, first isolated from porcine
creasing the blood flow through that capillary bed. Because brain in 1982, is structurally related to mammalian pan-
p-adrenoreceptors are rarely located near nerve endings, creatic polypeptide and peptide YY. Neuropeptide Y is
they usually are stimulated by circulating catecholamines. widespread throughout the animal kingdom and, so far,
Catecholamines are released into the bloodstream from has been identified in many vertebrates and in insects. Neu-
adrenergic neurons of the autonomic nervous system and ropeptide Y is co-localized with norepinephrine in sympa-
from chromaffin cells in the adrenal medulla. Circulating thetic ganglia and adrenergic nerves; it also is found in
catecholamines are dominated by epinephrine released many nonadrenergic fibers. The atrial and ventricular my-
from the adrenal medulla (see Chapter 8). Epinephrine re- ocardium and the coronary arteries are surrounded by
acts with both a- and p-adrenoreceptors, causing vasocon- nerve fibers that contain neuropeptide Y. In addition, it ap-
striction and vasodilation, respectively. Although a-adreno- pears that myocardial cells themselves can synthesize and
receptors are less sensitive to epinephrine, when activated secrete neuropeptide Y. In general, neuropeptide Y de-
they override the vasodilation mediated by p-adrenorecep- creases coronary blood flow and the contraction of cardiac
tors. The result is that high levels of circulating epinephrine muscle by reducing the level of inositol triphosphate
cause vasoconstriction and thus an increase in peripheral (InsP,), an intracellular second messenger (see Chapter 9).
CIRCULATION 511
......................................
Neuropeptide Y appears to ameliorate those actions of cat- Nitric oxide is produced and released continuously by
echolamines on the heart and coronary circulation medi- the vascular endothelium, causing relaxation of vascular
ated via InsP, .The role of neuropeptide Y in the peripheral smooth muscle. Nitric oxide-mediated vasodilator tone
circulation is less well understood, but it appears to ame- regulates blood flow and pressure in mammals and perhaps
liorate the increase in blood pressure resulting from other vertebrates. Observation of endothelium-dependent
norephinephrine-induced peripheral vasoconstriction me- vascular relaxation led to the discovery of endothelium-
diated by a-adrenoreceptors. derived relaxing factor (EDRF).It is now known that this
ATP, as well as neuropeptide Y, is stored and co- phenomenon results largely from the generation and release
released with catecholamines. ATP and its breakdown of nitric oxide, which activates guanyl cyclase, leading to
product, adenosine, act to inhibit release of catecholamines. an elevation of the intracellular second messenger cGMP
Adenosine is released by many tissues during hypoxia but (cyclic 3', 5' guanosine monophosphate). This compound
has only a paracrine or autocrine action because of rapid in turn mediates muscle relaxation.
inactivation. Hypoxia tends to promote catecholamine re- A family of enzymes, the nitric oxide synthases, oxidize
lease into the blood from chromaffin tissue, but this action L-arginine to nitric oxide and L-citrulline in the endothe-
is modulated by the local release of adenosine. lium. Several nitric oxide synthases are calcium dependent,
and calcium entry into endothelial cells has been shown to
Parasympathetic stimulation Arterioles in the circulation cause the production and release of nitric oxide and relax-
to the brain and the lungs are innervated by parasympa- ation of surrounding smooth muscle. The finding that some
thetic nerves. These nerves contain cholinergic fibers, which calcium channels in the endothelium are stretch sensitive
release acetylcholine from their nerve endings when stim- suggests that nitric oxide production in response to vessel
ulated. In mammals, parasympathetic nerve stimulation stretch may be due to increased calcium entry into the en-
causes vasodilation in arterioles. Some parasympathetic dothelium. A variety of chemicals (e.g., acetylcholine, ATP,
neurons release ATP and other purines from their endings. and bradykinin) stimulate release of nitric oxide, as does
Some of these purinergic neurons may participate in the hy; ,xis, pH change, and increased vessel shear stress.
control of capillary blood flow. ATP, for instance, causes Tht .:is evidence of increased production of nitric oxide
vasodilation. wit1 'wreasing pressure associated with each heartbeat.
h L r i coxide synthases have been found in a wide vari-
Local control of capillay blood flow ety of animals including horseshoe crabs, the blood-
Tissues require a basal capillary blood flow to supply nu- sucking bug Rhodnius, lampreys, and man. Nitric oxide
trients and 0, and to remove waste products. Active tissues has been shown to have many functions other than main-
have greater requirements and thus capillary blood flow taining vasodilator tone, so its presence in animals without
must increase during activity. In addition to nervous con- vascular tone or in nonvascular tissues is not surprising. For
trol of the central cardiovascular system, various mecha- example, nitric oxide released in the central nervous system
nisms control the microcirculation at the local level. For in- by stimulation of N-methyl-D-aspartate receptors is in-
stance, if a vessel is stretched by an increase in input volved in modulation of synaptic activity. Nitric oxide may
pressure, the vascular smooth muscle responds by con- also be involved in nonspecific defense reactions, the relax-
tracting, opposing any increase in vessel diameter. This ten- ation of nonvascular smooth muscle in the gastrointestinal
dency to maintain vessel diameter within narrow limits pre- and genito-urinary tracts, and the regulation of the release
vents large changes in resistance to flow and therefore of some hormones. In addition, nitric oxide released by en-
maintains a relatively constant basal flow through the cap- dothelial cells, platelets, and leukocytes modulates both cell
illary bed. Local heating of a tissue, which may accompany adhesion and aggregation and inhibits thrombosis.
inflammation, is associated with a marked vasodilation, The vascular endothelium releases endothelins and
whereas a reduction in temperature causes a vasoconstric- prostacyclin, as well as nitric oxide. Endothelins are small
tion. Thus an ice pack can reduce the blood flow and, there- vasoconstrictive proteins, containing 21 amino acid resi-
& fore, the swelling associated with damage to a tissue. dues. Prostacyclin causes vasodilation and acts as an an-
Numerous compounds also influence capillary blood ticoagulant. It thus functions as an antagonist of the
flow within a tissue. These can be grouped into three types: prostaglandin thromboxane A,, which promotes blood
compounds produced by the vascular endothelium; various clotting and causes vasoconstriction.
vasocontrictors and vasodilators released from other cells;
and metabolites associated with increased activity. Inflammatory and other mediators Thromboxane A, is
formed in the plasma from arachidonic acid released by
Endothelium-produced compounds The endothelium is platelets when they bind to damaged tissues. Although
not merely a barrier between blood and the underlying tis- thromboxane levels increase in a damaged tissue and
sues, but an active tissue, producing many compounds. cause vasoconstriction, local injury in mammals is ac-
Some of these, such as nitric oxide, endothelin, and prosta- companied by a marked vasodilation of vessels in the re-
cyclin, affect vascular smooth muscle and, therefore, capil- gion of the damage, due largely to the local release of his-
lary blood flow. tamine. Histamine is released, not from endothelial cells,
512 I N T E G R A T I O N OF P H Y S I O L O G I C A L SYSTEMS
......................................
but from some connective tissue and white blood cells in Although low 0, levels, indicative of tissue activity,
injured tissues. Antihistamines ameliorate, but do not cause vasodilation and increased blood flow in systemic
completely remove, this inflammatory response. Another capillaries, the lung capillary bed exhibits the opposite be-
group of potent vasodilators, plasma kinins, also are acti- havior. That is, low 0, in the lung causes local vasocon-
vated in damaged tissues. Tissue damage results in the re- striction rather than vasodilation. The functional signifi-
lease of proteolytic enzymes that split kininogen, an a2- cance of this difference relates to the direction of gas
globulin, into kinins. Hypoxia also stimulates formation transfer. In the lung capillaries, 0, is taken up by the blood,
of kinins. and thus blood flow should be greatest in the regions of
Among the vasocontrictors that act on arterioles are high 0,. In systemic capillaries, however, 0, leaves the
norepinephrine released from sympathetic nerves and an- blood for delivery to the tissues, and the highest blood flow
giotensin IT. Angiotensin is formed, primarily in the lungs, should be to the area of greatest need, which is indicated by
from angiotensinogen, which circulates in the blood (see regions of low 0,.
Chapter 14).Finally, serotonin acts as a vasoconstrictor or If blood flow to an organ is stopped by clamping the
vasodilator, depending on the vascular bed and on the dose artery or by a powerful vasoconstriction, there will be a
level. It is found in high concentration in the gut and blood much higher blood flow to that organ when the occlusion
platelets. is removed than there was before the occlusion. This phe-
Histamine, bradykinin, and serotonin cause an increase nomenon is termed reactive hyperemia. Presumably during
in capillary permeability. As a result, large proteins and the ischemic period (a period of no blood flow), O2 levels
other macromolecules tend to distribute themselves more are reduced, and CO,, H+, and other metabolites build
evenly between plasma and interstitial spaces, reducing up and cause a local vasodilation. The result is that when
the colloid osmotic pressure difference across the capillary the occlusion is removed, blood flow is much higher than
wall. Filtration thus increases, and tissue edema occurs. On normal.
the other hand, norepinephrine, angiotensin 11, and vaso-
pressin tend to promote absorption of fluid from the inter-
CARDIOVASCULAR RESPONSE
stitial fluid into the blood. This absorption can be achieved
TO EXTREME CONDITIONS
by reducing filtration pressure and/or the permeability of
the capillaries. In the previous sections, we've described the general orga-
nization of the circulation and its regulation under usual
conditions. The cardiovascular system responds in char-
acteristic ways during exercise, diving, and hemorrhage
to meet the physiological challenge of these extreme
conditions.
Exercise
Regulation of the cardiovascular system during exercise is
clearly a complex process involving central neural control
Metabolic conditions associated with activity When activ- mechanisms, peripheral neural reflex mechanisms (espe-
ity in a tissue increases, there must be a concomitant in- cially those involving skeletal muscle afferent fibers), and
crease in blood flow. Local control of capillary flow ensures local control. Many cardiovascular changes seen during ex-
that the most active tissue has the most dilated vessels and ercise can occur in the absence of neural mechanisms, indi-
therefore the most blood flow. The degree of dilation de- cating the importance of local control systems In increasing
pends on local conditions in the tissue, and those conditions blood flow to active skeletal muscles. The central neural
associated with high levels of activity generally cause va- control mechanisms and reflexes from muscle afferent
sodilation. The term hyperemia means increased blood mechanoreceptive and chemoreceptive inputs, however,
flow to a tissue; ischemia means the cessation of flow. clearly play a role, the exact form varying with the nature
Active hyperemia refers to the increase in blood flow that of the exercise. For example, the reflex effect on the car-
follows increased activity in a tissue, particularly skeletal diovascular system of muscle afferent stimulation depends
muscle. on the nature of the exercise:
Active tissues, metabolizing aerobically, are marked by
a decrease in 0, and an increase in CO, ,H+,various other Isometric contractions of muscles tend to raise blood
metabolites (e.g., adenosine, other ATP breakdown prod- pressure with little effect on cardiac output.
ucts), and heat. Extracellular K+ also rises in skeletal mus- Isotonic contractions raise cardiac output but cause lit-
cle following exercise. All of these activity-related meta- tle change in arterial blood pressure.
bolic changes, as well as nitric oxide and prostacyclin, have
been shown to cause vasodilation and a local increase in During exercise, blood flow to skeletal muscle is in-
capillary blood flow. That is, the most active tissue has the creased in proportion to the level of activity of the muscle.
most dilated vessels and therefore the highest blood flow. The increase in flow to a muscle may be as much as twenty
times; at the same time, transfer of oxygen from the blood changes in heart rate and little change in stroke volume dur-
to muscle may increase threefold, resulting in a sixtyfold ing exercise.
increase in oxygen utilization by the muscle. Active hyper- Exercise is associated with only small changes in arte-
emia is primarily responsible for increasing blood flow to rial blood pressure, pH, and gas tensions. The oscillations
muscle; the resulting decrease in peripheral resistance leads in PCO2and POI with breathing are somewhat larger, as is
to an increase in cardiac output mediated by sympathetic the arterial pressure pulse. The increased pressure pulse is
nerves. At the same time, there is a reduction in flow to dampened to some extent because of increased elasticity of
the gut, kidney, and, at high levels of exercise, the skin the arterial walls due to a rise in circulating catecholamines.
(Figure 12-46). Cardiac output can increase up to ten times It is probable that arterial chemoreceptors and barorecep-
above the resting level owing to large increases in heart rate tors play only a minor role in the cardiovascular changes
and small changes in stroke volume. Much of the increase associated with exercise. Motor neurons that innervate
in cardiac output can be accounted for by a decrease in pe- skeletal muscle are activated by higher brain centers in the
ripheral resistance to about 50% of the resting value and cortex at the onset of exercise (see Chapter 10); it is possi-
by an increase in venous return to the heart due to both the ble that this activating system also initiates changes in lung
pumping action of skeletal muscles on veins and the in- ventilation and blood flow. Proprioreceptive feedback from
crease in breathing associated with exercise. muscles may also play a role in increasing lung ventilation
The increased sympathetic, but decreased parasympa- and cardiac output (see Chapter 13). A number of other
thetic, activity in nerves innervating the heart has the effect changes augment gas transfer during exercise; for example,
of increasing both heart rate and the force of contraction, red blood cells are released from the spleen in many ani-
so as to maintain stroke volume at a relatively constant mals, increasing the oxygen carrying capacity of the blood.
level. In fact, stroke volume increases by about 1.5 times Thus, exercise is responsible for a complex series of inte-
during exercise in mammals, despite the large increase in grated changes that lead to delivery of adequate oxygen
heart rate and the associated reduced time available for fill- and nutrients to the exercising muscle.
ing and emptying. Following sympathetic stimulation,
however, blood is ejected more rapidly from the ventricles
with each beat, maintaining stroke volume at higher heart
rates. The relative role of changes in stroke volume and
heart rate in generating the increase in cardiac output with
exercise varies among animals. In fish, for example, the
changes in stroke volume are much greater than the
changes in heart rate, whereas in birds there are very large Diving
Many air-breathing vertebrates can remain submerged for
prolonged periods. During submersion for any period, all
Skin air-breathing vertebrates stop breathing, so the animal must
Heart, brain, etc.
rely on available oxygen stores in the blood (see Chapter
Viscera
13). The cardiovascular system is adjusted to meter out
the limited oxygen store to those organs-brain, heart,
and some endocrine structures-that can least withstand
anoxia.
Much of the information on the responses to submer-
sion has been collected from studies of animals forced to
dive, sometimes simply by holding an animal's head under
water. Because naturally occurring dives vary considerably
in depth, duration, and exercise level, information obtained
on forced dives is not always directly applicable to natural
dives. Whales and dolphins spend their lives in the water
going to the surface to breathe, whereas seals may spend
considerable time on land out of water. Other animals may
Max Vo, spend most of their time on land and dive only occasionally.
0,uptake (L . r n i n - I ) Oxygen stores vary in animals, so metabolism may be com-
Figure 12-46 During exercise, total cardiac output increases and blood pletely aerobic during some dives but largely anaerobic
flow shifts to the active muscles. Shown is the approximate distribution of during others.
cardiac output at rest and at different levels of exercise up to the maximal Figure 12-47 illustrates the typical cardiovascular
oxygen consumption (Max Vo2)in a normal young man. The progressive
changes that occur when a seal dives and remains sub-
reduction in the absolute blood flow and percentage of cardiac output
distributed to the viscera (splanchnicregion and kidneys) augments mus-
merged. In mammals, but not in other vertebrates, stimu-
cle blood flow. Even skin is constricted during brief periods of exercise at lation of the facial receptors that inhibit breathing cause a
high oxygen consumption. [Adapted from Rowell, 1974.1 marked bradycardia. Although the initial pressurization of
514 I N T E G R A T I O N O F P H Y S I O L O G I C A L SYSTEMS
......................................
I Predive + Dive 4 Recovery is low before a dive, there may be little or no change in
heart rate during the dive. If the heart rate is high, then
there may be a marked bradycardia due to wetting the face
and a decrease in lung stretch-receptor activity.
The "water" receptors present in birds are not directly
involved in the cardiovascularchanges associated with sub-
mersion. A decrease in heart rate is not observed either in
submerged ducks breathing air through a tracheal cannula
or in submerged ducks following carotid body denervation
(Figure 12-48).Thus, activation of the "water" receptors
causes suspension of breathing (apnea); the subsequent
drop in blood Pol and pH and the rise in PCOlresult in stim-
ulation of chemoreceptors, which then reflexly cause the
cardiovascular changes.
Stimulation of lung stretch receptors in mammals mod-
oxygen ifies the reflex response initiated by chemoreceptor stimu-
content lation. In the absence of breathing, and hence stimulation
I I
of lung stretch receptors, different reflex responses are
Time --+ elicited by chemoreceptor stimulation than when the ani-
Figure 12-47 The cardiovascular system undergoes numerous adjust- mal is breathing. In the absence of breathing, lung inflation
ments when a seal dives. Heart rate, cardiac output, and blood O2con- tends to suppress the reflex cardiac inhibition and periph-
tent decrease during a dive, but blood CO, content increases. During the eral vasoconstriction caused by stimulation of arterial
recovery period after a d~ve,blood lactate increases greatly; the other pa-
chemoreceptors. As a submerged animal rises in the water
rameters first overshoot and then gradually return t o predive values.
column, the lung becomes inflated, possibly activating
stretch receptors in the lung and causing cardiac accelera-
the lung can lead to a transient increase in blood O2 and tion. When the animal is breathing, stimulation of arterial
CO, levels, the continued utilization of 0, during the dive chemoreceptors results in a marked increase in lung venti-
results in a gradual fall in blood 0, and rise in blood CO, lation. In this case, low blood 0, andlor high blood CO,
levels. This fall in blood 0, stimulates the arterial chemore- levels cause peripheral vasodilation. This vasodilation leads
ceptors and, in the absence of lung stretch-receptor activity, to an increase in cardiac output to maintain arterial pres-
causes peripheral vasoconstriction and a reduction in heart sure in the face of increased peripheral blood flow. Thus,
rate and cardiac output; thus blood flow to many tissues is the hypoxia (low oxygen) caused by cessation of breathing
reduced so as to maintain flow to the brain, heart, and during a dive is associated with bradycardia and a reduc-
some endocrine organs.
The absence of lung stretch-receptor activity is due to
the absence of breathing and the compression of the lung
as the animal descends in the water column. The increase
A Control
in peripheral resistance results from a marked rise in sym-
pathetic output and involves constriction of fairly large ar- In Out Rate
teries. Reductions in blood flow to the kidney have been
recorded in Weddell seals during a dive. In some instances,
blood flow to muscle decreases, but this depends on the
level of exercise associated with the dive and the species.
Sometimes arterial pressure rises during a dive, causing
B After carotid body denervation -
stimulation of arterial baroreceptors; in such dives brady- 4-
observed that mice lived longer if plant material was present duction in animals and found that the amount of heat produced
in their containers. Priestley's observations caused Benjamin relative to oxygen uptake was about the same for animals and
Franklin to remark that the practice of cutting down trees near burning coal, although the rates of these processes were much
houses should cease as plants were able to restore air, which is slower in animals.
spoiled by animals. Thus Priestley demonstrated that plants, as Lavoisier was also a tax collector. Such people are generally
well as certain chemicals when heated, could produce some gas not held in high repute and this brilliant scientist was no excep-
that keeps animals and flames alive. He thought this gas could tion: he was sent to the guillotine in 1794.
absorb phlogiston, something that was released when material
of blood between the tissues and the respiratory epithe- - Figure 13-1 schematically illustrates the components of
lium. Blood flows through an extensive capillary network the gas-transfer system in many animals, which involves
and is spread in a thin film just beneath the respiratory four basic steps:
surface, thereby reducing diffusion distances required to
distribute the contained gases. The gases are transported 1. Breathing movements, which assure a continual sup-
between the respiratory surface and the tissues by bulk ply of air or water to the respiratory surface (e.g., lungs
flow of blood in the circulatory system. Gases diffuse be- or gills)
tween blood and tissues across the capillary wall. Once 2. Diffusion of 0, and C 0 2 across the respiratory
again, to facilitate gas transfer, the area for diffusion is epithelium
large, and the diffusion distance between any cell and the 3. Bulk transport of gases by the blood
nearest capillary is small. Graham's law states that the rate 4. Diffusion of 0, and C 0 2 across capillary walls be-
of diffusion of a substance along a given gradient is in- tween blood and mitochondria in tissue cells
versely proportional to the square root of its molecular
weight (or density). Because oxygen and carbon dioxide The capacity of each of these steps is matched because natural
molecules are of similar size, they diffuse at similar rates in selection tends to eliminate metabolically costly unutilized ca-
air; they are also utilized (02 ) and produced ( C 0 2) at ap- pacities. This matching of capacities in a chain of linked
proximately the same rates by animals. It can therefore be events has been referred to as symmorphosis.Presumably the
expected that a transfer system that meets the oxygen re- capacities of elements in a chain will be determined by the ca-
quirements of an animal will also ensure adequate rates of pacity of the rate-limiting step. Capacities in a chain of events,
carbon dioxide removal. however; are not always matched, and symmorphosis draws
GAS E X C H A N G E A N D A C I D - B A S E B A L A N C E 519
.............................................................................
Diffusion Diffusion Figure 13-1 The gas-transport system of a verte-
I
brate consists of two pumps and two diffusion bar-
I
I
I
I
I
I
1 : - in series between the external envi-
riers alternatinq
ronment and the tissues. [Adapted from Rahn, 1967.1
Air or water
Fluid Blood
Pump Pump
attention to these apparently uneconomic design features. OXYGEN AND CARBON DIOXIDE
One explanationfor over- or under-capacity is that a single el- IN BLOOD
ement can be a link in several chains; thus its capacity may be
appropriate for one chain of events, but be in excess for an- In considering the movement of oxygen and carbon diox-
other, explaining the apparent excess capacity. ide between the environment and the cells, we will first dis-
The rate of flux of gases varies enormously among cuss how these gases are transported in the blood, rather
animals, from 0.08 ml.g-l-h-l in an earth worm to than starting with either the environment or the cell. We
40 ml .g - l hpl in a hovering hummingbird. The concentra- take this approach because the mechanisms by which oxy-
tions of aerobic enzymes (e.g., cpochrome oxidase) and gen and carbon dioxide are carried in the blood affects their
cristae area per mitochondrion both increase with metabolic transfer between the environment and the blood and the
rate. The hummingbird and some insects, however, may have blood and the tissues.
reached the upper limit for rates of oxygen utilization by an-
Respiratory Pigments
imals. Clearly mitochondrial volume and density in muscles
cannot be increased indefinitely without compromising the Once oxygen diffuses across the respiratory epithelium into
capacity of the muscles to contract; that is, there must be some the blood, it combines with a respiratory pigment that gives
relationship between structures that supply energy (mito- the characteristic color to the blood. The best known res-
chondria) and structures that use energy (myofilaments).The piratory pigment, hemoglobin, is red. By binding oxygen,
space occupied by mitochondria never exceeds 45% of the to- the respiratory pigment increases the 0, content of blood.
tal volume in muscle, even in mammals, birds, and insects, the In the absence of a respiratory pigment, the 0, content of
animals with the highest oxygen uptakes. There must also be blood would be low. The Bunsen solubility coefficient of
limits to mitochondrial design in terms of the number of oxygen in blood at 37°C is 2.4 m l 0 , per 100 ml of blood
cristae per unit mitochondria1volume, the ultimate miniatur- per atmosphere of oxygen pressure. Therefore, the concen-
ization being determined by the minimum volume required tration of 0, in physical solution (i.e., not bound to a res-
by the enzymes involved in energy production. It would seem piratory pigment) in human blood at a normal arterial Po2
that hummingbirds, and perhaps some other small mammals will be only 0.3 m l 0 , per 100 ml blood, or 0.3 vol % 0,.
and a few insects, may have approached these limits of design In fact, the total 0, content of human arterial blood at a
determining the maximum rates of oxygen uptake. normal arterial Po, is 20 vol %. The 70-fold increase in
Insects are usually much smaller than the smallest birds content is due to the combination of oxygen with hemo-
and mammals. Some large insects seem to have been dis- globin. In most animals using hemoglobin as a respiratory
placed by small birds, rather like the monoplane displacing pigment, the 0, content in physical solution is only a small
the biplane before World War 11. Vertebrate mini'aturization fraction of the total 0, content of the blood. The Antarctic
may be limited by the nature of their gas-transfer systems. icefish is an exception among the vertebrates; the blood of
Insects have a tracheal system that exchanges gases directly this fish lacks a respiratory pigment and therefore has a low
between the medium and the tissues, permitting high rates 0, content. It compensates for the absence of hemoglobin
of oxygen uptake in very small animals. with an increased blood volume and cardiac output, but its
rate of 0, uptake is reduced compared with that of species
from the same habitat that have hemoglobin. Low temper-
atures probably are a factor in the evolution of fishes lack-
ing hemoglobin. Low temperatures are associated with low
metabolic rates in poikilotherms, and oxygen, like all gases,
has a higher solubility at low temperatures.
Respiratory pigments are complexes of proteins and
metallic ions, and each one has a characteristic color. The
520 I N T E G R A T I O N O F P H Y S I O L O G I C A L SYSTEMS
...............................................................................
SPOTLIGHT 13-2 water, but as temperature decreases, water will condense, and
thls condensation WIII also reduce the explred gas volume Ifthe
THE GAS LAWS barometric pressure IS 760 mm ~ gand
, the water vapor pressure
at 37" and 20°C is 47 mm Hg and 17 mm Hg , respectively, then
Over 300 years ago, Robert Boyle determined that at a glven a measured gas volume at 20°C of 500 ml IS converted to BTPS
temperature the product of pressure and volume IS constant for explred volume as follows
a glven number of molecules of gas Gay-Lussac's law states
that elther the pressure or the volume of a gas IS d~rectlypro-
500 ml X
(760 - 17) (273 + 37) = 551 ml
portlonal to absolute temperature ~fthe other IS held constant (760 - 47) (273 + 20)
Combined, these laws are expressed In the equat~onof state
for a gas: Thus, under the conditions stated above, a gas volume of 551 ml
within the lung is reduced to 500 ml following exhalation be-
cause of the drop in gas temperature and the condensation of
water.
where P is pressure, Vis volume, n is number of molecules of a Dalton's law of partial pressure states that the partial pres-
gas, R is the universal gas constant (0.08205 L.atm. K-I. mot-', sure of each gas in a mixture is independent of other gases pre-
or 8.314 x lo7ergs. O K - ' .mol-', or 1.987 cal . K-' . mol-I), and sent, so that the total pressure equals the sum of the partial pres-
Kis absolute temperature. For accurate use, the equation should sures of all gases present. The partial pressure of a gas in a
be modified by using van der Waals constants. mixture will depend on the number of molecules present in a
The equation of state for a gas indicates that equal volumes given volume at a given temperature. Usually, oxygen accounts
of different gases atthe same temperature and pressure contain for 20.94% of all gas molecules present in dry air; thus, if the to-
equal numbers of molecules (Avogadro's law). One mole of gas tal pressure is 760 mm Hg, the partial pressure of oxygen, Po*,
occupies approximately 22.414 liters at O°C and 760 mm Hg. Be- will be 760 x 0.2094 = 159 mm Hg. But air usually contains wa-
cause the number of molecules per unit volume is dependent on ter vapor, which contributes to the total pressure. If the air is 50%
pressure and temperature, the conditions should always be saturated with water vapor at 22"C, the water vapor pressure is
stated along with the volume of gas. Gas volumes in physiology 18 mm Hg. If the total pressure is 760, the partial pressure of oxy-
are usually reported as being at body temperature, atmospheric gen will be (760 - 18) x 0.2094 = 155 mm Hg. If the partial
pressure, and saturated with water vapor (BTPS);at ambient tem- pressure of CO, in a gas mixture is 7.6 mm Hg and the total pres-
perature and pressure, saturated with water vapor (ATPS); or at sure is 760 mm Hg, then 1% of the molecules in air are CO,.
standard temperature and pressure (O°C, 760 mm Hg) and dry, Gases are soluble in liquids. The quantity of gas that dis-
or zero water vapor pressure (STPD). solves at a given temperature is proportional to the partial pres-
Gas volumes measured under one set of conditions (e.g., sure of that gas in the gas phase (Henry's law). The quantity of
ATPS) can be converted to another (e.g., BTPS) by using the gas in solution equals a P, where Pis the partial pressure of the
equation of state for a gas. For example, the volume of air ex- gas and a is the Bunsen solubility coefficient, which is inde-
pired from a mammalian lung at a body temperature of 37OC pendent of P. The Bunsen solubility coefficient varies with the
(273 + 37 = 310 K) is often measured at ambient room temper- type of gas, the temperature, and the liquid in question, but is
+
ature, say 20°C (273 20 = 293 K).The drop in temperature will constant for any one gas in a given liquid at constant tempera-
reduce the expired gas volume. A gas in contact with water will ture. The Bunsen solubility coefficient for oxygen decreases with
be saturated with water vapor. The water vapor pressure at 100% increases in ionic strength and temperature of water.
saturation varies with temperature. Expired air is saturated with
color of a respiratory pigment changes with its 0, content. into four subunits of approximately equal weight, each
Thus, hemoglobin, which is bright red when it is loaded containing one polypeptide chain and one heme group.
with 02,becomes a dark maroon-red when deoqgenated. Myoglobin, a respiratory pigment that stores 0, in verte-
Vertebrate hemoglobin, except that of cyclostomes, has brate muscles, is equivalent to one hemoglobin subunit and
a molecular weight of 68,000 and contains four iron- exhibits considerable sequence homology with the hemo-
containing porphyrin prosthetic groups, called heme, asso- globin a chain.
ciated with globin, a tetrameric protein (Figure 13-2A).The In a hemoglobin molecule, iron in the ferrous state
globin molecule consists of two dimers, alp, and alp,, (Fe2+)is bound into the porphyrin ring of the heme, form-
each of which is a tightly cohering unit. The two dimers are ing coordinate links with the four pyrrole nitrogens
more loosely connected to each other by salt bridges, ex- (Figure 13-2B). The two remaining coordinate linkages
cept that the two p chains do not touch. Oxygenation are used to bind the heme group to an 0, molecule and
alters these bridges, leading to conformational changes in to the imidazole ring of a histidine residue in the globin
the hemoglobin molecule. Hemoglobin can be dissociated (Figure 13-2C). If O2is bound, the molecule is referred to
as oxyhemoglobin; if 0, is absent, it is called deoxyhemo-
globin. Binding of 0, to hemoglobin to form oxyhemoglo-
bin does not oxidize ferrous to ferric iron. Oxidation of the
ferrous iron in hemoglobin to the ferric state produces
methemoglobin, which does not bind 0, and therefore is
nonfunctional. Although formation of methemoglobin oc-
curs normally, red blood cells contain the enzyme methe-
moglobin reductase, which reduces methemoglobin to the
functional ferrous form. Certain compounds (e.g., nitrites
and chlorates) act either to oxidize hemoglobin or to inac-
tivate methemoglobin reductase, thereby increasing the
level of methemoglobin and impairing oxygen transport.
The affinity of hemoglobin for carbon monoxide is
about 200 times greater than its affinity for oxygen. As a re-
sult, carbon monoxide will displace oxygen and saturate
hemoglobin, even at very low partial pressures of carbon
monoxide, causing a marked reduction in oxygen transport
to the tissues. Hemoglobin saturated with carbon monox-
ide is called carboxyhemoglobin.The effect of such satura-
tion on oxidative metabolism is similar to that of oxygen
deprivation, which is why the carbon monoxide produced
by cars or improperly stoked coal or wood stoves is so
toxic. Even the levels found in city traffic can impair brain
function owing to partial anoxia.
Hemoglobin is found in many invertebrate groups, but
others possess different respiratory pigments, including
hemerythrin (Priapulida, Brachiopoda, Annelida), chloro-
cruorin (Annelida), and hemocyanin (Molluscs, Arthrop-
OOC oda). Many invertebrates do not have a respiratory pig-
Heme
ment. Hemocyanin, a large, copper-containingrespiratory
pigment, has many properties similar to those of hemoglo-
C bin, binding oxygen when the partial pressure is high and
releasing it when the partial pressure is low. Hemocyanin
binds oxygen in the ratio of 1mol of 0, to approximately
L-Histidine
(His) 75,000 g of the respiratory pigment. In comparison, 4 mol
COO- of 0, bind to 68,000 g of hemoglobin when it is completely
I saturated. Unlike hemoglobin, hemocyanin is not packaged
+H,N-C-H
I in cells and is not associated with high levels of carbonic an-
hydrase in the blood. In its oxygenated form, it is light blue;
in its unoxygenated form, it is colorless.
heme groups for oxygen. As a hemoglobin molecule is oxy- of oxygen at which the hemoglobin is 50% saturated with
genated, it goes through a conformational change from the oxygen; the lower the P,, ,the higher the oxygen affinity. As
tense (T) state to the relaxed (R) state. Oxygenation is as- the curves in Figure 13-3 demonstrate, myoglobin has a
sociated with changes in the tertiary structure near the much higher oxygen affinity than hemoglobin. Variations
hemes that weaken or break connections between the a, PI in oxygen affinity among hemoglobins are related to dif-
and a, p, dimers, leading to a large change in the quater- ferences in the protein globin, not to differences in the heme
nary structure from the T to the R state. These conforma- group. Each a and P chain of the globin molecule consists
tional changes also produce changes in the dissociation of of between 141 and 147 amino acids, depending on the
acidic side chains, so that protons (H+ions) are released as chain and the hemoglobin in question. The amino acid se-
hemoglobin is oxygenated. -I quences of both the a and the P chains from different
An important property of respiratory pigments is that hemoglobins exhibit many similarities, but there are some
they combine reversibly with 0, over the range of partial differences. Although most amino acid substitutions are
neutral, some have a marked impact on function. For ex-
ample, a genetic defect resulting in substitution of valine for
glutamic acid in position 6 of the p chain causes human he-
moglobins to form large polymers that distort the erythro-
cyte into a sickle shape, giving rise to sickle cell anemia. Be-
cause these sickle cells cannot pass through small blood
vessels, oxygen delivery to tissues is impaired. Individuals
with both normal and sickle cell hemoglobins suffer only
mild debilitation but have greater resistance to malaria,
thus ensuring the continuation of the sickle cell gene in the
population. Certain amino acids in globin bind various li-
gands, and substitution of these residues can cause changes
in the oxygen affinity of hemoglobin.
The rate of oxygen transfer to and from blood increases
in proportion to the difference in Po, across an epithelium.
A hemoglobin with a high oxygen affinity facilitates the
movement of 0, into the blood from the environment be-
cause 0, is bound to hemoglobin at low Po?;i.e., 0, enter-
ing the blood is immediately bound to hemoglobin, so 0,
Figure 13-3 Hemoglobinswith multiple heme groups have sigrnoid oxy- is removed from solution and Po2is kept low. Thus, a large
gen dissociation curves, whereas myoglobin with only a single heme
difference in PO2is maintained across the respiratory ep-
I,
group has a hyperbolic dissociation curve. Lamprey hemoglobin, with a
single heme group, has a dissociation curve similar to that of myoglobin. ithelium-and therefore a high rate of oxygen transfer into
P,, the partial pressure at which a respiratory pigment is 50% saturated the blood-until hemoglobin is fully saturated. Only then
L with oxygen, is a measure of its oxygen affinity. does blood Po2rise. Hemoglobin with a high oxygen affin-
GAS EXCHANGE A N D ACID-BASE BALANCE 523
......................................
ity, however, will not release O2 to the tissues until the Po> throcytic organophosphate differs among species. For in-
is very low. In contrast, a hemoglobin with a low oxygen stance, mammalian erythrocytes contain high levels of 2,3-
affinity will facilitate the release of 0, to the tissues, main- diphosphoglycerate (DPG);indeed, hemoglobin and DPG
taining large differences in Pol between blood and tissues are nearly equimolar in human erythrocytes. DPG binds to
and a high rate of oxygen transfer to the tissues. Thus, a he- specific amino acid residues in the p chains of deoxyhe-
moglobin of high oxygen affinity favors the uptake of 0, moglobln, but DPG binding decreases with increasing pH.
by the blood, whereas a hemoglobin of low oxygen affinity Increases in DPG levels accompany reductions in blood 0,
facilitates the release of 0, to the tissues. From a functional or hemoglobin concentrations, increases in pH, or both.
viewpoint, therefore, hemoglobin should have a low O2 Low blood O2 levels may result from a climb to a higher
affinity in the tissues and a high 0, affinity at the respira-
tory surface. In light of this, it is highly significant that the
oxygen affinity of hemoglobin is affected by changes in
Arterial
chemical and physical factors in the blood that favor oxy-
gen binding at the respiratory epithelium and oxygen re-
lease in the tissues.
The hemoglobin-oxygenaffinity is labile and dependent
on the conditions within the red blood cell. For instance,
the hemoglobin-oxygena f f i t y is reduced by the following:
Elevated temperature
Binding of organic phosphate ligands including 2,3-
diphosphoglycerate (DPG),ATP, or GTP
Decrease in pH (increase in H+ concentration)
Increase in CO,
"
30 60 90 '
The hemoglobin molecule has a much higher affinity for Po, (mm Hg)
ligands when it is in the T, or deoxygenated, state.
Figure 13-4 The oxygen affinity of hemoglobin decreases with decreas-
Increases in H+ concentration (decreases in pH) cause
ing pH. Because of this phenomenon, called the Bohr effect, changes
a reduction in the oxygen affinity of hemoglobin, a in blood Pco,, wh~chinfluence blood pH, indirectly affect hemoglobin-
phenomenon termed the Bohr effect, or Bohr shift oxygen affinity. Shown are experimental blood oxygen dissociation
(Figure 13-4). Carbon dioxide reacts with water to form curves in humans at three pH values. The Po,values of mixed venous
carbonic acid and reacts with -NH2 groups on plasma and arterial blood are indicated. [Adapted from Bartels, 1971.]
proteins and hemoglobin to form carbamino compounds.
Thus an increase in PCo2causes a reduction in the oxygen
affinity of hemoglobin in two ways: by decreasing blood
pH (Bohr effect) and by promoting the direct combination
of CO, with hemoglobin to form carbamino compounds.
Therefore, when C 0 2 enters the blood at the tissues it fa-
cilitates the unloading of 0, from hemoglobin, whereas
when CO, leaves the blood at the lung or gill, it facilitates
the uptake of 0, by the blood. The oxygen dissociation
curve for myoglobin, unlike that for hemoglobin, is rela-
tively insensitive to changes in pH.
Hemocyanins from the Dungeness crab, Cancer mag-
ister, and some other invertebrates exhibit a Bohr shift sim-
ilar to that of hemoglobin (Figure 13-5). But hemocyanins
from several gastropods and from the horseshoe crab,
Limulus, show a greater oxygen affinity with a decrease in
pH. This phenomenon, referred to as a reverse Bohr effect,
may facilitate oxygen uptake during periods of low oxygen
availability when prolonged reductions in blood pH,occur 30 60 90
in these animals. Po, (mm Hg)
As noted above, the binding of organic phosphate
Figure 13-5 Some hemocyanins, like hemoglobin, exhibit a Bohr shift.
compounds to hemoglobin reduces the oxygen affinity of Blood oxygen dissociat~oncurves for the crab Cancer magistershown
most vertebrate hemoglobins, except those from cy- here indicate that the hemocyanin from this crab shows a Bohrshift. [Un-
clostomes, crocodiles, and ruminants. The dominant ery- published data supplied by D. G. McDonald.]
524 INTEGRATION OF PHYSIOLOGICAL SYSTEMS
.......................................
altitude, as both barometric pressure and the partial through the solution; the gradient for the deoxygenated
pressure of 0, in air decrease with altitude. The resultant pigment is in the reverse direction from that for oxygen and
DPG rise in humans in response to high altitude is com- the oxygenated pigment. Hence, the oxygenated pigment
pleted in 24 hours with a half-time of about 6 hours. At el- diffuses in the same direction as oxygen, whereas the de-
evations of 3000 m, the DPG concentration in erythro- oxygenated pigment diffuses in the reverse direction. Thus,
cytes is 10% greater than it is at sea level. The low 0, a pigment such as hemoglobin may facilitate the mixing of
levels at altitude result in a decrease in blood O2 levels, gases in the blood, and myoglobin may play a similar role
which stimulates breathing. The resulting increase in ven- within tissues.
tilation (i.e., exchange of air between the lungs and ambi-
ent air) reduces C0, levels in the blood and raises blood
pH, which increases hemoglobin-oxygen affinity. The ele-
vation in DPG at altitude offsets the effects of reduced CO,
levels and maintains hemoglobin-oxygen affinity close to
that at sea level.
In the erythrocytes of some vertebrates, other phos-
phorylated compounds are present In hlgher concentra-
tion than DPG, and consequently they play a more im-
portant role in modulating the oxygen affinity of In some fishes, cephalopods, and crustaceans, an in-
hemoglobin than does DPG. In most fishes ATP and/or crease in CO, or a decrease in pH causes not only a reduc-
GTP has this function, whereas inositol pentaphosphate tion in the oxygen affinity of hemoglobin but also a reduc-
(InsP, ) is the dominant erythrocytic organophosphate in tion in oxygen capacity, which is termed the Root effect, or
birds. In the Amazonian fish Arapazrna gzgas, ATP is the Root shift (Figure 13-6). In those hemoglobins showing a
dominant erythrocytic organophosphate in the young Root shift, low pH reduces oxygen binding to hemoglobin,
aquatic form, but InsP, is dominant in the obligate air- so that even at high Po2,only some of the binding sites are
breathing adult. oxygenated; that is, 100% saturation is never achieved.
Pho~phor~lated compounds in the erythrocyte not only An increase in temperature exacerbates problems of
affect the oxygen affinity of hemoglobin but also increase oxygen delivery in poikilothermic aquatic animals such as
the magnitude of the Bohr effect and may affect subunit fishes. A rise in temperature not only reduces oxygen solu-
interaction. It appears that in mammals the functional sig- bility in water but also decreases the oxygen affinity of he-
nificance of increased DPG levels is to maintain hemoglo- moglobin, making oxygen transfer between water and
bin-oxygen affinity under hypoxic (low-oxygen) condi- blood more difficult. Unfortunately, this decrease in affin-
tions, as at high altitude. In contrast, hypoxia reduces ity occurs at a time when tissue oxygen requirements are in-
erythrocytic organophosphate levels in fishes. In these creasing, also as a result of the rise in temperature.
animals, however, hypoxia is often associated with a de- It is generally assumed that a particular hemoglobin has
crease in blood pH (acidosis),rather than the increase in evolved to meet the special gas-transfer and H+ buffering
pH (alkalosis) seen in mammals at altitude. The effect of requirements of the animal. Differences in the properties of
the reduction in ATP (or GTP) in fishes is to offset the ef- hemoglobins are due to variation in the amino acid se-
fects of this hypoxia-associated acidosis, thereby maintain- quence of the peptide chains of the globin portion of the
ing blood-oxygen affinity. Thus in a functional sense the ef-
fects of changing erythrocytic organophosphate levels are
similar in both fishes and mammals; in both instances the
result is to maintain hemoglobin-oxygen affinity.
Reaction velocities for the binding of oxygen to hemo-
globin are rapid and usually do not limit rates of oxygen
transfer. The rate at which oxygen can bind to hemoglobin,
however, also depends on the hemoglobin concentration.
The higher the hemoglobin concentration the more oxygen
bound per unit time. The more oxygen bound per unit time Plasma
the longer the persistence of a large diffusion gradient only
across the respiratory epithelium for oxygen and, therefore,
the higher the rate of oxygen transfer.
The presence of a respiratory pigment also increases the
transfer of oxygen through the blood, because the oxy- Figure 13-6 Redua~onsIn pH decrease the blood oxygen capac~ty(Root
effect) In hemoglob~ns from some teleost f~shesThese oxygen e q u ~ l ~ b -
genated pigment co-diffuses with oxygen down the con- rlum curves of eel blood were obta~nedat 14°Cw ~ t hthe pH from 6 99 to
centration gradient. That is, a gradient exists for both oxy- 8 20 The bottom llne descr~besthe 0, content of plasma [Adapted from
gen and the oxygenated pigment in the same direction Steen, 1963.1
GAS E X C H A N G E A N D A C I D - B A S E B A L A N C E 525
......................................
molecule, the heme portion being the same in all hemoglo-
bins. Not only do hemoglobins vary among species, but
they also may change during development. In humans, for
example, several genes encode plike globin chains, and the
relative expression of these chains differs during prenatal
and postnatal life (Figure 13-7).Human fetal hemoglobin,
which contains y chains, rather than adult P chains, has a
higher 0, affinity than adult hemoglobin. The higher 0,
affinity of fetal hemoglobin enhances oxygen transfer from
mother to fetus. As the proportion of fetal hemoglobin de-
creases and adult hemoglobin increases following birth, the
oxygen affinity of the blood decreases (Figure 13-8).Other
mammals exhibit similar differences between fetal and
adult hemoglobins.
It is important to remember that in most animals he-
moglobin is contained within red blood cells, but the val-
ues of blood parameters usually refer to conditions in the
plasma, not in the red blood cell. Differences in these pa-
rameters exist between the inside and outside of cells, in- Po, (mm Hg)
cluding red blood cells. For example, the pH of mammalian Figure 13-8 In humans, the oxygen affinity of blood decreasesfor about
arterial blood at 37°C is usually 7.4. This is the pH of ar- three months after birth as the fetal hemoglobin is replaced by adult he-
terial blood plasma; the pH inside the red blood cell is less, moglobin (see Figure 13-7).These blood oxygen dissociation curves were
about 7.2 at 37OC. determined at a pH of 7.40. [Adapted from Bartels, 1971.1
Carbon Dioxide Transport in Blood Carbon dioxide also reacts with hydroxyl ions to form
bicarbonate:
Carbon dioxide diffuses into the blood from the tissues, is
transported in the blood, and diffuses across the respiratory
surface into the environment. Carbon dioxide reacts with
=H+ + OH-
H,O
water to form carbonic acid, a weak acid, which dissociates CO, + OH- =HCO,
into bicarbonate and carbonate ions:
The proportion of CO,, HC0,-, and C0:- in solution
CO, + H,O ----T H,CO, H + + HC0,- depends on pH, temperature, and the ionic strength of the
solution. In mammalian blood at pH 7.4, the ratio of CO,
HC0,- H + + C0;-
to H2C03is approximately 1000: 1, and the ratio of CO,
to bicarbonate ions is about 1: 20. Bicarbonate is, there-
fore, the predominant form of CO, in the blood at normal
blood pH. The carbonate content is usually negligible in
/ Fetal birds and mammals; in poikilotherms, however, with their
hemoglobin low temperature and high blood pH, the carbonate content
(y-chain) may approach 5% of the total CO, content of the blood,
1 (p-chain)
I but bicarbonate is still the predominant form of CO,.
9k Carbon dioxide also reacts with -NH2 groups on pro-
teins and, in particular, hemoglobin to form carbamino
: Embryonic compounds.
:hemoglobin \ A, (&chain)
: (€-chain)
protein -NH, + CO, H+ + protein --NHCOO-
., The extent of carbamino formation depends on the num-
3 6 Birth 3 6
Duration of pregnancy Age ber of available terminal NH, groups, and it increases with
(months) (months) blood pH and increasing CO, levels. The terminal NH,
groups of both the a and P chains of mammalian, bird, and
Figure 13-7 Hemoglobins change during development in humans. The
reptile hemoglobins are available for carbamino formation.
relative amounts of the various hemoglobin p-l~kechains synthesized in
the fetus changes during the course of pregnancy. Fetal hemoglobin,
The terminal NH, group of the a chain of fish and am-
which contains two a and two y chains, has a h~gheroxygen affinity than phibian hemoglobins, however, is acetylated and therefore
adult hemoglobin [Adapted from Young, 1971.] not available for carbamino formation. Because
organophosphates bind to some of the same amino acids
that are involved in carbamino formation, organophos- Deoxygenated blood,
phate binding reduces carbamino formation. However,
high pH reduces organophosphate binding and so aug-
ments carbamino formation by making more NH, groups
available. Because fish erythrocytes often have high
organophosphate levels as well as acetylated a chains, fish
rely less on carbamino formation for CO, transport than
mammals.
The sum of all forms of CO, in the blood-that is,
molecular C 0 2 , H,CO, ,HC0,-, COj2-, and carbamino
compounds-is referred to as the total CO, content of the
blood. The CO, content varies with PCO2, and the relation-
ship can be described graphically in the form of a CO, dis-
sociation curve (Figure 13-9).As Pco2increases, the major
change is in the bicarbonate content of the blood. The for-
mation of bicarbonate is, of course, pH dependent. The re- Figure 13-9 The total CO, content of blood increaseswith Pco,, but only
lationships between plasma H C O , concentration and the volume of molecular CO, increases linearly. Note that at a glven Pco,
plasma pH at three values of PCo2are shown graphically in oxygenated blood contains less C 0 2 than deoxygenated blood
(Haldane effect). A and V refer t o arterial and venous blood levels,
Figure 13-10. A decrease in pH at constant PCo2is associ-
respectively.
ated with a fall in bicarbonate. The pH of red blood cells is
less than that of plasma, but PCo2is in equilibrium across
the cell membrane. Therefore, bicarbonate levels are lower HC0,- or reacts with-NH, groups of hemoglobin and
in erythrocytes than in plasma. Erythrocytes usually con- other proteins to form carbamino compounds. The reverse
stitute less than 50% of the blood volume (i.e., plasma process occurs when CO, is unloaded from the blood. The
volume is greater than erythrocyte volume), and the bicar- largest change is In the HC0,- concentration; changes in
bonate concentration is higher in plasma than in erythro- the levels of CO, and carbamino compounds usually rep-
cytes; it thus follows that most of the bicarbonate in the resent less than 20% of total carbon dioxide excretion.
blood is in plasma. The reaction of CO, with O H to form H C O , is slow
and has an uncatalyzed time course of several seconds. But
Transfer of Gases to and from the Blood in the presence of the enzyme carbonic anhydrase, this re-
As CO, is added to the blood in the tissues and removed at action approaches equilibrium In much less than a second.
the respiratory surface, the levels of CO,, HC0,-, and car- Although plasma has a higher total CO, content than red
bamino compounds all change. Carbon dioxide both enters blood cells, most of the CO, entering and leaving the
and leaves the blood as molecular C0, rather than as bi- plasma does so via erythrocytes, because carbonic anhy-
carbonate ion because CO, molecules diffuse through drase is present in red blood cells but not in the plasma.
membranes much more rapidly than HC0,- ions. In the Therefore, formation of H C O , ions in the tissues and CO,
tissues, CO, enters the blood and either is hydrated to form in the lungs occurs predominantly in red blood cells; once
-z-
E
a 30 -
poventilatlon, then plasma pH and blcarbonate are al-
tered beyond the normal range, as ~nd~cated by the
whole-body buffer l~ne [Adapted from Davenport,
C
m -28
0 19741
%.-
0
- HC0,-
a -23
m
20-
-
m
a
- 2
Whole-body
buffer l ~ n e
I I I I I I I I
lol 710 7.2 7.4 7.6 7.8
Plasma pH
GAS E X C H A N G E A N D A C I D - B A S E B A L A N C E 527
...............................................................................
formed, H C O , ions and CO, subsequently are transferred which can occur in either direction, bicarbonate flowing
from or into the plasma. out of the erythrocyte in the tissues and into the erythrocyte
On entering the blood from the tissues, CO, diffuses at the respiratory surface (Figure 13-llB).Band I11 protein
into red blood cells, and HC0,- is formed rapidly in the is present in all vertebrate erythrocytes except those of lam-
presence of carbonic anhydrase (Figure 13-11A). As the prey and hagfish. In these animals bicarbonate stays within
HC0,- level within erythrocytes rises, H C O , ions move the red blood cell and there is no anion transfer between the
from the cells into the plasma. Electrical balance within the erythrocyte and the plasma.
cells is maintained by anion exchange; as HC0,- ions leave A second reason why most of the CO, entering or leav-
the red blood cells, there is a net influx of C 1 ions from the ing the blood passes through the red blood cells is that oxy-
plasma into the cells, a process called the chloride shift. Red genation of hemoglobin (Hb) causes H + release, thereby
blood cells, unlike many other cells, are very permeable to acidifying the cell interior; conversely, deoxygenation re-
both C 1 and H C O , because the membrane has a high sults in the binding of H+to Hb. Thus 0, binding to Hb at
concentration of a special anion carrier protein, the band the respiratory surface facilitates the formation of CO,,
III protein. This transport protein binds C1- and HC0,- whereas release of 0, from Hb in the tissues facilitates the
and transfers them in opposite directions through the ery- formation of H C O , (Figure 13-12).As a result, changes
throcyte membrane. The anion exchange is passive and de- in pH associated with the transfer of CO, into or out of the
pends on concentration gradients to drive the process, blood are minimized because of proton binding to and
COP TISSUE
T Capillary
I 1 wall
C02 + H20 =
(slow)
H2C03L=;HCO, + H+
T I
Figure 13-11 Most of the carbon d~oxideentering the blood In the tls-
sues and leavlng the blood In the lungs passes through red blood cells
(A) Carbon dloxlde produced in the tlssues rapldly forms blcarbonate
(HC0,-) In the red blood cell because the hydration reactlon 1s catalyzed
by carbonlc anhydrase present In the cell B~carbonateleaves the eryth-
rocyte In exchange for chloride, and excess protons are bound by de-
oxygenated hemoglobin (Hb) (B) These reactions are reversed In lungs
Oxygen enterlng the red blood cell displaces protons from Hb, and car-
bon d ~ o x ~ denters
e the plasma Carbonlc anhydrase (lndlcated by solld
clrcles) In the membrane of the lung endothellal cells converts some of
the plasma blcarbonate to carbon dloxlde Movement of carbon dloxlde
across the respiratory surface IS augmented by the dlffus~onof blcarbon-
ate and ~ t sconversion back to carbon dloxlde at the outer surface, a
process termed facll~tateddiffusion
528 I N T E G R A T I O N O F P H Y S I O L O G I C A L SYSTEMS
.........................................
Capillary Respiratory CO, can occur at the catalyzed rate in plasma as blood per-
endothelium epithelium fuses the lung capillaries (see Figure 13-11B).In addition,
oxygenation of hemoglobin acidhes erythrocytes in the lung
capillaries, facilitating the conversion of HC0,- to CO,,
which then diffuses into the plasma and across the lung ep-
ithelium. The resulting decrease in erythrocyte bicarbonate
levels results in the influx of HC0,- ions from the plasma
accompanied by the outward movement of C1- ions. The
relative quantities of HC0,- converted to CO, in the ery-
throcytes and the plasma of the blood perfusing the respi-
ratory epithelium is influenced by the extent of proton pro-
duction associated with hemoglobin oxygenation and the
OH- lco2+> amount of carbonic anhydrase activity in the walls of the
respiratory epithelium. In teleost fish, for instance, the plasma
perfusing the gills is not exposed to carbonic anhydrase. In
Figure 13-12 The pH changes associatedwith the changes in blood Pco,
these animals, most excretion of CO, occurs through the red
in the tissues and respiratorysurface are offset by binding and release of
H+ ions by deoxygenated and oxygenated blood. For example, transfer
blood cells and is tightly coupled to 0, uptake through pro-
of CO, into the blood in the tissues causes a decrease in pH due to for- ton production by oxygenation of hemoglobin.
mation of bicarbonate; concomitant deoxygenation of hemoglobin frees Carbonic anhydrase activity is also found on the en-
proton acceptors, which bind the excess H+ ions. The opposite reactions dothelial surfaces of a number of systemic capillary beds,
occur at the respiratory epithelium. including those in skeletal muscle. In these capillaries, for-
mation of HC0,- catalyzed by carbonic anhydrase can oc-
proton release from hemoglobin as it is deoxygenated and cur in the absence of red blood cells. Thus some of the CO,
oxygenated, respectively. transferred into the blood in skeletal muscle does not pass
For example, as the PC,> increases in the tissues, the through erythrocytes. Carbonic anhydrase also facilitates
subsequent formation of H C O , or carbamino compounds carbon dioxide transfer, referred to as facilitated diffusion
liberates H + ions. At the same time, release of oxygen of CO, (see Figure 13-llB), which results from the simul-
forms deoxyhemoglobin, which binds protons. As taneous diffusion through the epithelium of bicarbonate
deoxygenation proceeds, however, more proton acceptors and protons, the latter also augmented by release from
become available on the hemoglobin molecule. In fact, buffers. Carbonic anhydrase catalyzes the rapid intercon-
complete deoxygenation of saturated hemoglobin, releas- version of CO, and H C O , in this process of facilitated dif-
ing 1 mol O,, results in the binding of 0.7 mol of H+ ions. fusion, with CO, entering and leaving the cell.
Thus, when the ratio of CO, production to 0, consump- There are at least seven forms of carbonic anhydrase,
tion (calledthe respiratory quotient) is 0.7, the transport of designated CA-I through CA-VII. All are similar in struc-
CO, can proceed without any change in blood pH. (As dis- ture and catalyze the interconversion of carbon dioxide and
cussed in Chapter 16, the respiratory quotient depends on bicarbonate. Carbonic anhydrase I (CA-I)and carbonic an-
the type of diet.) Even when the respiratory quotient is 1, hydrase I1 (CA-11),present in human red blood cells, have
the additional 0.3 mol H + is buffered by blood proteins, in- a molecular weight of about 29,000, containing about 260
cluding hemoglobin, and blood undergoes only a small amino acid residues. CA-11, an extremely efficient catalyst
change in pH. At a given PCo1, deoxyhemoglobin binds of the carbon dioxide-bicarbonate hydration-dehydration
more protons, thereby facilitating H C O , formation, and reactions, is found in a wide variety of tissues, including the
reacts with CO, to form carbamino hemoglobin more eas- brain, eye, kidney, cartilage, liver, lung, pancreas, gastric
ily than does oxyhemoglobin. As a result, the total CO, mucosa, skeletal muscle, and anterior pituitary, as well as
content of deoxygenated blood at a given Pco, is higher red blood cells. This form is involved in a wide variety of
than that of oxygenated blood (seeFigure 13-9).Thus, de- functions, augmenting the supply of bicarbonate and/or
oxygenation of hemoglobin in the tissues reduces the protons for a number of cellular and metabolic processes.
change in PCol and pH as CO, enters the blood; this is A few humans exhibit an inherited CA-I1 deficiency, the
termed the Haldane effect. pattern of inheritance being autosomal recessive. Although
In the lungs, two mechanisms are available for transfer these individuals have no detectable CA-11, they have nor-
of CO, from the blood. As noted already, carbonic anhy- mal levels of CA-I in their red blood cells. CA-I1 deficiency
drase is absent from plasma, and thus the interconversion of not only compromises the gas-exchange process but also
CO, and HC0,- occurs at the slow, uncatalyzed rate in produces many other symptoms including metabolic aci-
plasma. (Any carbonic anhydrase liberated by the break- dosis, renal tubular acidosis, and sometimes mental retar-
down of red blood cells is excreted via the kidney.) In the en- dation. In addition, because CA-I1is involved in production
dothelial cells of lung capillaries, however, carbonic anhy- of protons needed for bone resorption in osteoclasts, its ab-
drase is embedded in the cell surface, accessible to plasma sence results in osteoporosis, often associated with multi-
CO, and HC03-. Therefore, the conversion of H C O , to ple bone fractures. The wide range of symptoms associated
with inherited CA-I1 deficiency reflects the large number of
functions in which CA-I1plays a role in augmenting proton
and/or bicarbonate delivery.
The rate of movement of CO, and 0, into or out of the C 0 .
.-0
red blood cell is determined by the diffusion distance and the d-4
2 100 ~orse
diffusion coefficient of these substances through the cell. (U
Temperature ("C)
25 30 35
OH-
Extracellular fluid
+ CO,
II I
20 min
Increase in
extracellular CO,
I
Overshoot
7.5 -
CO, influx
CO, efflux
7.0 -
Slow H' efflux
Increased
NH,CI in
extracellular
fluid
Figure 13-16 Changes in extracellular carbon dioxide and ammonium a gradual rise in the intracellular pH. (B) If extracellular NH,CI levels rise
chloride levels cause changes in intracellularpH of tissue cells. (A) If CO, sharply, NH, diffuses rapidly into the cell and combines with hydrogen
levels in the extracellular fluid are suddenly increased, CO, diffuses ions to form ammonium ions, which diffuse slowly across the cell mem-
rapidly into the cell, forming bicarbonate and causing a sharp fall in in- brane (dashed line). As a result, the intracellular pH increases.
tracellular pH. A subsequent slow efflux of H+ ions (dashed line) leads to
ple, some cells can actively pump protons out via a proton
ATPase in the membrane; this proton efflux can result in a
sodium influx. Often acid extrusion is accompanied by
chloride efflux, presumably in exchange for extracellular
HC03-, which has been shown to be required for pH reg-
ulation by cells. For instance, the drug SITS (4-acetamido-
4'-isothiocyanostilbene-2,2'-disulfonicacid), which blocks
chloride-bicarbonate exchange in erythrocytes, also in-
hibits pH regulation in other cells.
Thus both proton-exchange and anion-exchange mech-
anisms in the cell membrane play an important role in ad-
justing intracellular pH. An acid load in the cell is accom-
panied by H+ efflux coupled to Na+ influx and by HC0,-
influx coupled to C 1 efflux. The movement of HC0,- Figure 13-17 The Jacob-Stewart cycle is the cycling of carbon dioxide
and bicarbonate to transfer acid between the extracellular and intracel-
into the cell is equivalent to movement of H+ out of the cell
lular compartments. In a red blood cell, depicted here, the cycle gener-
because H C O , ions that enter the cell are converted to ally operates to transfer acid from the plasma to the cell interior. Because
CO, ,releasing hydroxyl ions and increasing pH. The CO, carbonic anhydrase is present only inside cells, the slow, uncatalyzed in-
so formed, leaves the cell and is converted to bicarbonate, terconversion of CO, and HC0,- in the extracellularfluid determines the
releasing protons. This cycling of CO, and HC03-, re- rate of acid transfer.
ferred to as the Jacobs-Stewart cycle, functions to transfer
H + ions from the cell interior in the face of an intracellular
acid load, such as that generated by anaerobic metabolism membrane, and the membrane potential maintains a
(Figure 13-17). lower pH inside the red blood cell than in the plasma. A
In most vertebrate red blood cells, unlike most other sudden addition of acid to the plasma (e.g., following
cells, hydrogen ions are passively distributed across the anaerobic production of H + )results in a fall in erythrocyte
534 I N T E G R A T I O N O F P H Y S I O L O G I C A L SYSTEMS
.........................................
pH. The acid is transferred from the plasma to the interior fact, catecholaminesreleased into the blood during periods
of the erythrocyte not by diffusion of H+ ions but by of metabolic acidosis activate the erythrocyte Na+/H+ex-
bicarbonate-chloride exchange (seeFigure 13-17).The ad- changer, which moves H+ out of and Na+ ions into the cell.
dition of H + to the plasma causes PCo1to increase due to In fish with a large muscle mass, burst swimming results in
the conversion of HC0,- to CO,, which then diffuses into a marked acidosis. This drop in plasma pH, if transferred
the red blood cell and is converted to HC03-, thereby re- to the red blood cell, would impair oxygen binding to he-
ducing intracellular pH. Bicarbonate then diffuses out of moglobin and reduce the ability of the fish to swim aero-
the cell via the chloride-bicarbonate exchange mechanism. bically. This does not happen because erythrocytic pH, in
Thus in erythrocytes, the Jacobs-Stewart cycle operates to these fish is regulated and remains high during the acido-
transfer acid from the plasma to the cell interior. sis following burst-swimming activity.
muscle can and does tolerate much larger oscillations in increases even more in mammals, the total effect being an
pH. As a result, the brain has extensive, if poorly under- increase in respiratory surface area per unit volume of lung.
stood, mechanisms for regulating the pH of the cere- In general, the area of the respiratory surface in mammals
brospinal fluid (CSF). In the face of a sudden acid load in increases with body weight and the rate of oxygen uptake
the blood, hydrogen ions are taken up by the muscles, re- (Figure 13-20). Teleost fishes typically have a smaller res-
ducing oscillations in the blood and protecting the brain piratory surface area than mammals of equivalent body
and other more sensitive tissues. Hydrogen ions are then weight.
slowly released into the blood from muscle and excreted ei- The mammalian lung consists of millions of blind-
ther via the lungs as CO, or via the kidney in acid urine. ending, interconnected sacs, termed alveoli. The trachea
Thus, when there is a sudden acid load in the body, the subdivides to form bronchi, which branch repeatedly, lead-
muscles can act as a temporary H+reservoir, thus reducing ing eventually to terminal bronchioles and finally respira-
the magnitude of the oscillations in pH in other regions of tory bronchioles, each of which is connected to a terminal
the body. spray of alveolar ducts and sacs (Figure 13-21).The total
Figure 13-19 The different gas-transfer sys-
Air tems in air-breathing and water-breathing
animals are associated with characteristic
1 AIR
distribution of respiratory gases in the blood
and tissues. (A) Schematic diagrams of 0, and
\
CO, flows in air-breathing and water-breath-
ing animals. (B) Relative values of Po2 and Pcq
Blood
in the inhalant medium, blood, and tissues in
air-breathing (top) and water-breathing (bot-
tom) animals.
+==r
Capillaries
c02
Tissue
Water
Slope = 1.0
loo
10
Mammals 7 1000
Human
Monkey
Figure 13-20Respiratory surface area increases w~thsize (A) Relation- (S. A.) and oxygen uptake in mammals. [Part A adapted from Randall,
ship between resp~ratorysurface area and body weight In selected mam- 1970; part B from Tenney and Ternmers, 1963.1
mals and teleost fishes. (6) Relat~onshipbetween alveolar surface area
GAS E X C H A N G E A N D ACID-BASE BALANCE 537
...............................................................................
keeps the lungs clean (see Chapter 8). In the respiratory
portions of the lung, smooth muscle replaces cartilage.
Contraction of this smooth muscle can have a marked ef-
fect on the dimensions of the airways in the lungs.
Small mammals have a higher resting 0, uptake rate
per unit body weight than large mammals because of their
greater alveolar surface area per unit body weight. This in-
crease in respiratory surface area is achieved by a reduction
in the size but an increase in the number of alveoli per unit
volume of lung. In humans the number of alveoli increases
rapidly after birth, the adult complement of about 300 mil-
lion being attained by the age of eight years; subsequent in-
creases in respiratory area are achieved by increases in the
volume of each alveolus. The resting 0, uptake rate per
unit weight is higher in children than in adults; once again
there is a correlation between uptake per unit weight and
alveolar surface area per unit body weight.
The diffusion barrier in mammals is made up of an
aqueous surface film, the epithelial cells of the alveolus, the
interstitial layer, endothelial cells of the blood capillaries,
plasma, and the wall of the red blood cell (Figure 13-22B).
\
Several cell types compose the lung epithelium. Type I cells,
\
\
Respiratory the most abundant, constitute the major part of the lung
\ bronchiole
epithelium. They are squamous epithelial cells, having a
thin platelike structure, a single cell extending into the two
adjacent alveoli with the nucleus tucked away in a corner.
Type I1 cells are characterized by a laminated body within
the cell and have surface villi; type I1 cells produce surfac-
tants, discussed later. Type 111cells are mitochondrion-rich
cells with a brush border. These rare cells appear to be in-
bronchiole
volved in NaCl uptake from lung fluid. In addition to these
cells, a number of alveolar macrophages wander over the
surface of the respiratory epithelium. It is generally as-
Figure 13-21 In the mammalian lung, a series of branching, progres- sumed, but not demonstrated, that the coefficient of diffu-
sively smaller ducts deliver air to the respiratory portion, consisting of ter- sion for gases does not vary in the lungs of different ani-
minal and respiratory bronchioles and alveolar duas and sacs. Gas trans-
fer occurs across the respiratory epithelium shown in red.
mals, the only structural variables being lung area and
diffusion distance between air and blood.
The following terms are used to describe different types
of breathing and lung ventilation:
cross-sectional area of the airways increases rapidly as a re-
sult of extensive branching, although the diameter of indi- Eupnea-normal, quiet breathing typical of an animal
vidual air ducts decreases from the trachea to the terminal at rest.
bronchioles. The terminal bronchioles, the respiratory
bronchioles, the alveolar ducts, and the alveolar sacs con- Hyperventilation and hypoventilation-increase and
stitute the respiratory portion of the lung. Gases are trans- decrease, respectively, in the amount of air moved in or
ferred across the thin-walled alveoli found in the regions out of the lung by changes in the rate and/or depth of
distal to the terminal bronchioles, termed acini. The air- breathing, such that ventilation no longer matches CO,
ways leading to the terminal bronchioles constitute the production and blood CO, levels change
nonrespiratory portion of the lung. Alveoli in adjoining Hyperpnea-increased lung ventilation due to increase
acini are interconnected by a series of holes, the pores of in breathing in response to increased CO, production
Kohn, allowing the collateral movement of air, which may
be a significant factor in gas distribution during lung ven-
tilation (Figure 13-22A).
. (e.g., during exercise)
Apneahabsence of breathing
Dyspnea-labored breathing associated with the un-
Air ducts leading to the respiratory portion of the lung
pleasant sensation of breathlessness
- and a little smooth muscle and are lined
contain cartilage
with cilia. The epithelium secretes mucus, which is moved Polypnea-increase in breathing rate without an in-
toward the mouth by the cilia. This "mucus escalator" crease in the depth of breathing
A Figure 13-22 During ventilation of the mammalian lung,
the respiratory gases move to and from the alveolar space
and blood in the pulmonary capillaries. (A)Three interalve-
Alveolus olar septa of dog lung meeting at junction line. Connective
tissue fibers lie in the central plane, forming a continuous
tensile network with which the capillary network is interwo-
ven. Endothelial cellsand type I epithelial cellsform the lin-
ing ofthe thin air-blood barrier. Pores of Kohn connect alve-
Capillary
oli. (B) Dimensions and structure of the alveolar capillary
membrane. [Part A adapted from Weibel, 1973; part B from
Hildebrandt and Young, 1965.1
Alveolus B
Alveolus
- 5 ~m
lining
(0.01 ~ r n )
space
(0.02-0.2 pm)
Air exchanged between the alveoli and the environ- tion, leaving a residual volume of air in the lungs. The max-
ment must pass through a series of tubes (trachea, bron- imum volume of air that can be moved in or out of the
chi, nonrespiratory bronchioles) not directly involved in lungs is referred to as the vital capacity of the lungs. These
gas transfer. At the end of exhalation the air contained and other terms used to describe various volumes and
in these tubes will have come from the alveoli and will capacities associated with lung function are illustrated in
be low in oxygen and high in carbon dioxide. This air Figure 13-23.
will be the first to move back into the alveoli at the next The 0, content is lower and the CO, content is higher
breath. At the end of inhalation the nonrespiratory in alveolar gas than in ambient air because only a portion
tubes will be filled with fresh air and this volume will be of the lungs' gas volume is changed with each breath.
~ -
the first to be exhaled with the next breath. Thus this vol- Alveolar ventilation in humans is about 350 ml, whereas
ume is not involved in gas transfer and, therefore, is the functional residual volume of the lungs exceeds 2000
referred to as the anatomical dead space volume. Some air ml. During inspiration
- the ducts leading to the alveoli
-
-
may be supplied to nonfunctional alveoli, or certain alve- elongate and widen, causing an increase in acinar volume.
oli may be ventilated at too high a rate, increasing the During breathing, air moves in and out of the acinus and
volume of air not directly involved in gas exchange. This may also move between adjacent alveoli through the
volume of air, termed the physiological dead space, is usu- pores of Kohn. Mixing of gases in the ducts and alveoli
ally greater than, but includes, the anatomical dead space occurs by diffusion and by convection currents caused by
(Spotlight 13-3). breathing (Figure 13-24 ). In the alveolar ducts, 0, dif-
The amount of air moved in or out of the lungs with fuses toward the alveoli and CO, away from them. Partial
each breath is referred to as the tidal volume. The amount pressures of 0, and CO, are probably fairly uniform
of fresh air moving in and out of the alveolar air sacs equals across the alveoli, because diffusion is rapid in air and the
the tidal volume minus the anatomical dead-space volume, distances involved are small. The partial pressures of gases
and is referred to as the alveolar ventilation volume. Only within the alveoli oscillate in phase with the breathing
this gas volume is directly involved in gas transfer. The movements, the magnitude depending on the extent of
lungs are not completely emptied even at maximal expira- tidal ventilation.
539
-
GAS E X C H A N G E A N D A C I D - B A S E B A L A N C E
...............................................................................
Figure 13-23 Numerous terms are used to de-
Maximum scribe various volumes and capacities associated
inspiratory with lung function. The t~dalvolume is the volume of
level air typically moved In and out of the lung, whereas
reserve
the vital capacity is the maximum volume.
Vital
capacity
Resting end-
inspiratory
-
-level
Resting end-
expiratory
level
C
I
f ~unctional Maximum
Total expiratory
lung level
volume capacity
I
Figure 13-25 The extremely long trachea of the trumpeter swan results
in a large anatomical dead-space volume. For comparison, see Figure
13-29 illustrating the length of the human trachea. 1From Banko, 19Ml.l
VA = VT - v,
But VA = V, - V
,, so substituting into this equation, we obtain Thus the physiological dead space of the lungs can be calcu-
lated from measurements of tidal volume, V,, and the CO, par-
- VD) + (PICO2X V
PEC02X VT = PACOZ(VT ), tial pressures in arterial blood, P,CO, and expired air, PEC02.
the heart. In the vertical (upright)human lung, arterial pres- The pulmonary vessels are very distensible and subject
sure is just sufficient to raise blood to the apex of the lung; to distortion by breathing movements. Small vessels within
hence flow is minimal at the top and increases toward the the interalveolar septa are particularly sensitive to changes
base of the lung (Figure 13-27).Blood is distributed more in alveolar pressure. The diameter of these thin-walled col-
evenly to different parts of the horizontal lung. lapsible capillaries is determined by the transmural pressure
Blood flow
(arterial blood pressure within capillaries, Pa,minus alveo- lated and blood flow will be high. Although pulmonary hy-
lar pressure, PA). If the transmural pressure is negative (i.e., poxic vasoconstriction is important in directing blood flow
PA> Pa),these capillaries collapse and blood flow ceases. to well-ventilated regions of the lung, it leads to problems
This collapse may occur at the apex of the vertical human when animals are exposed to general hypoxia, as may oc-
lung, where Pa is low (see Figure 13-27).If pulmonary ar- cur at high altitudes (see later section).
terial pressure is greater than alveolar pressure, which in Cardiac output to the pulmonary circuit is identical to
turn is greater than pulmonary venous pressure, then the cardiac output in the systemic circuit in mammals and
difference between arterial and alveolar pressure will de- birds. In amphibians and reptiles, with a single or partially
termine the diameter of capillaries in the interalveolar septa divided ventricle that ejects blood into both the pulmonary
and, in the manner of a sluice gate, control blood flow and the systemic circulation, the ratio of pulmonary to sys-
through the capillaries. Venous pressure will not affect flow temic blood flow can be altered. In turtles and frogs, there
into the venous reservoir as long as alveolar pressure ex- is a marked increase in blood flow to the lung following a
ceeds venous pressure. Flow in the upper portion of the ver- breath due to pulmonary vasodilation. During periods be-
tical lung is probably determined in this way by the differ- tween breaths in the frog Xenopus, pulmonary blood flow
ence between arterial blood pressure and alveolar pressure. decreases, but systemic blood flow is hardly changed pos-
Arterial blood pressure (and therefore blood flow) increases sibly because the ventricle is undivided (Figure 13-28).
with distance from the apex of the lung. These animals breathe intermittently, and variable blood
In the bottom half of the vertical lung, where venous flow to the gas exchanger, independent of blood flow to the
pressure exceeds alveolar pressure, blood flow is deter- rest of the body, permits some control of the rate of oxygen
mined by the difference between arterial and venous blood use from the lung store and rapid renewal of blood oxygen
pressures. This pressure difference does not vary with po- stores during ventilation. In addition, cardiac work is re-
sition, although both the arterial and venous pressures in- duced during apnea.
crease toward the base of the lung. This increase in absolute
pressure results in an expansion of vessels and, therefore, Ventilation of the Lung
a decrease in resistance to flow. Thus, flow increases toward The mechanism of lung ventilation varies considerably
the base of the lung, even though the arterial-venous pres- among animals. These variations reflect differences in the
sure difference does not change (seeFigure 13-27).The po- functional anatomy of the lungs and associated structures.
sition of the lungs with respect to the heart is therefore an First we will see how the mammalian lung is ventilated
important determinant of pulmonary blood flow. The lungs and then consider ventilation in birds, reptiles, frogs, and
surround the heart, thus minimizing the effect of gravity on invertebrates.
pulmonary blood flow as an animal changes from a hori-
zontal to vertical position. This close proximity of lungs
and heart within the thorax also has significance for cardiac
function: the reduced pressures within the thorax during in-
m Breathing movements, buccal cavity
halation aid venous return to the heart. This is often re-
ferred to as the thoraco-abdominal pump.
Even though the mammalian pulmonary circulation
lacks well-defined arterioles, both sympathetic adrenergic Blood flow, right pulmocutaneous
and parasympathetic cholinergic fibers innervate the
smooth muscle around the pulmonary blood vessels and 45
bronchioles. The pulmonary circulation, however, has E 30E
much less innervation than does the systemic circulation l5 B I O O ~pressure, left pu~mocutaneous
and is relatively unresponsive to nerve stimulation or in-
Time (min)
jected drugs. Sympathetic nerve stimulation or the injection
of norepinephrine causes a slight increase in resistance to
- I I
blood tlowv, whereas parasympathetic nerve stimulation or
acetylcholine has the opposite effect.
- A
,
L
0.1 [
0
Reductions in either oxygen levels or pH cause local B I O O ~ flow, left systemic
I I Residual volume lung volume and pressure within the thorax when muscles
are relaxed, but the glottis is closed. V, is the lung volume
when alveolar pressure is the same as ambient pressure and
the lung-chest system is relaxed. The points Iand E repre-
sent the pressure and volume of the system following inspi-
Alveolar pressure - ambient pressure (mm Hg) ration and expiration during quiet breathing.
544 I N T E G R A T I O N O F PHYSIOLOGICAL SYSTEMS
.........................................
(Figure 13-31A). Contractions of the diaphragm account tube, the mesobronchus, which joins the trachea anteriorly
for up to two-thirds of the increase in pulmonary volume. (Figure 13-33A). The parabronchi and connecting tubes
The increase in thoracic volume reduces alveolar pressure, form the lung, which is contained within a thoracic cavity.
and air is drawn into the lungs. Relaxation of the di- A tight horizontal septum closes the caudal end of the tho-
aphragm and external intercostal muscles reduces thoracic racic cage. The ribs, which are curved to prevent lateral
volume, thereby raising alveolar pressure and forcing air compression, move forward only slightly during breathing;
out of the lungs (Figure 13-31B).During quiet breathing, as a result, the volume of the thoracic cage and lung
pulmonary volume between breaths is at an intermediate changes little during breathing. The large flight muscles of
value, V,, at which the alveolar and ambient pressures are birds are attached to the sternum and have little influence
equal (see Figure 13-30).Under these conditions exhalation on breathing. Although there is no mechanical relation be-
often is passive, simply due to relaxation of the diaphragm tween flight and respiratory movements in birds, "in
and external intercostals. With increased tidal volume, ex- phase" flight and breathing movements may result from
piration becomes active, owing to contraction of the inter- synchronous neural activation of the two groups of muscles
nal intercostal muscles, which further reduces thoracic vol- involved.
ume until it drops below V, at the end of expiration. How, then, is the avian lung ventilated?The answer lies
in the associated air-sac system connected to the lungs (see
Birds Figure 13-32).As these air sacs are squeezed, air is forced
In birds, gas transfer takes place in small air capillaries through the parabronchi. The system of air sacs, which ex-
(10 pm in diameter) that branch from tubes called tend as diverticula of the airways, penetrates into adjacent
parabronchi (Figure 13-32). The functional equivalent of bones and between organs, reducing the density of the bird.
mammalian alveolar sacs, parabronchi are a series of small Of the many air sacs, only the thoracic (cranial) and ab-
tubes extending between large dorsobronchi and ventro- dominal (caudal) sacs show marked changes in volume
bronchi, both of which are connected to an even larger during breathing. Volume changes in the air sacs are
achieved by a rocking motion of the sternum against the
A Inhalation vertebral column and by lateral movements of the posterior
CJL-, ribs. Air flow is bidirectional in the mesobronchus, but
unidirectional through the parabronchi (Figure 13-33B).
Vertebral During inspiration, air flows into the caudal air sacs
column through the mesobronchus; air also moves into the cranial
Rib air sacs via the dorsobronchus and the parabronchi. Dur-
Rib
movement ing expiration, air leaving the caudal air sacs passes
Diaphrag
intercostal Extdrnal
1 through the parabronchi and, to a lesser extent, through
the mesobronchus to the trachea. The cranial air sacs,
whose volume changes less than that of the caudal air sacs,
are reduced somewhat in volume by air moving from the
cranial sac via the ventrobronchi to the trachea during
intercostal expiration.
Oxygen diffuses into the air capillaries from the
Exhalation parabronchi and is taken up by the blood. The air in the
parabronchi is changed during both inspiration and expi-
P-7 ration, enhancing gas transfer in the bird lung. The unidi-
rectional flow is achieved not by mechanical valves but by
aerodynamical valuing. The openings of the ventrobronchi
and dorsobronchi into the mesobronchus show a variable,
direction-dependent resistance to air flow. The structure of
the openings is such that eddy formation, and therefore re-
sistance to flow, varies with the direction of air flow.
Reptiles
Internal The ribs of reptiles, like those of mammals, form a thoracic
intercostal
cage around the lungs. During inhalation, the ribs are
~xternal
intercostal
i J t~ moved cranially and ventrally, enlarging the thoracic cage.
As this expansion reduces the pressure within the cage be-
movement low atmospheric pressure, and the nares and glottis are
Figure 13-31 The volume of the thorax increases during inhalation
open, air flows into the lungs. Relaxation of muscles that
(A) and decreases during exhalation (B) in mammals due to movement enlarge the thoracic cage releases energy stored in stretch-
of the ribs and diaphragm. ing the elastic component of the lung and body wall, al-
GAS E X C H A N G E A N D A C I D - B A S E B A L A N C E 545
......................................
Cervical
Lungs sacs lnterclavicular
Figure 13-32 In bird lungs, gas exchange occurs in air capillaries ex- ing tubes form the lung. During breathing, volume changes occur in the
tending from parabronchi, small tubelikestructuresthat are the functional associated air sacs, not in the thoracic cage and lungs. [Photograph cour-
equivalent of alveoli in mammals. The parabronchi (right) and connect- tesy of H. R. Duncker.]
lowing passive exhalation. Although reptiles do not possess the buccal cavity with the nares open and glottis closed;
a diaphragm, pressure differences between the thoracic and then the nares are closed, the glottis is opened, and the buc-
abdominal cavities have been recorded, indicating at least cal floor raised, forcing air from the buccal cavity into the
a functional separation of these cavities. lungs (Figure 13-34).This lung-filling process may be re-
In tortoises and turtles, the ribs are fused to a rigid shell. peated several times in sequence. Expiration also may be a
The lungs are filled by outward movements of the limb step process, the lungs releasing air in portions to the buc-
,
flanks andlor the plastron (ventral part of the shell) and by cal cavity. Expiration may not be complete, so that some of
forward movement of the shoulders.The reverse process re- the air from the lung is mixed with ambient air in the buc-
sults in lung deflation. As a result, retraction of limbs and cal cavity and then pumped back into the lungs. That is, a
head into the shell leads to a decrease in pulmonary volume. mixture of pulmonary air, presumably low in 0, and high
in CO,, is mixed with fresh air in the buccal cavity and re-
Frogs turned to the lungs. The reason for this complex method of
In frogs, the nose opens into a buccal cavity, which is con- lung ventilation is not clear, but it may be directed toward
nected via the glottis to paired lungs. The frog can open and reducing oscillations in CO, levels in the lungs in order to
close its nares and glottis independently. Air is drawn into stabilize and regulate blood Pco2 and control blood pH.
B
Dorsobronchi Parabronchi
Abdominal
sac
Cervlcal
sac r,- .-L
lnterclavicular /
sac /'ble~-
~ r a c h ~ a bronchus
\u'
) ~ d s tsac
thoracic
Trachea
Prethoracic
sac
Figure 13-33 Squeezing of airs sacs forces air through the parabronchi to the mesobronchus. (B) Schematic diagram of airflowthrough the bird
in bird lungs. ( A ) T ~ &
avian bronchial tree and associated air sacs. The air lung. Flow in the parabronchi is unidirectional.Solid arrows represent flow
sacs of the cranial group (cervical, interclavicular, and prethoracic sacs) during inspiration; open arrows, flow during expiration. [Adapted from
depart from the three cranial ventrobronchi, whereas the air sacs of the Scheid et al., 1972.1
caudal group (postthoracic and abdominal sacs) are connected directly
In aquatic snails, the lungs serve to reduce the animal's
density.
Pulmonary Surfactants
The lung wall tension depends on the properties of the alve-
olar wall and the surface tension at the liquid-air interface.
Surface tension is the force that tends to minimize the area
of a liquid surface, causing liquid droplets to form a sphere.
It also makes a surface film resistant to stretch, so that work
must be done to stretch a liquid surface. Because the alve-
oli are so compliant, the surface tension of their liquid lin-
ing contributes about 70% of the lung wall resistance to
Buccal Buccal plus stretch. If the liquid lining was just water, the alveolar wall
movement lung movement tension would be much higher than in fact it is, and large
forces would be required to inflate the lung and to separate
membranes glued together by surface tension. The expla-
nation for the relatively low surface tension of the liquid
lining the lungs is the presence of surfactants, lipoprotein
Buccal complexes that bestow a very low surface tension on the
volume liquid-air interface. Lung surfactants not only reduce the ef-
fort associated with breathing but also help prevent col-
lapse of alveoli.
Open glottis losed glottis Surfactants are produced by type I1 cells within the
- alveolar lining and have a half-life of about 12 hours
Nares closed in mammals. The predominant lipid in these lipo-
protein complexes is dipalmitoyl lecithin. The lipoprotein
film is stable, the lipid forming an outer monolayer firmly
associated with the underlying protein layer. Synthesis
- 0.5 s
pressure
of surfactants requires cortisol, and their release can
be stimulated by sighing. Surfactants are found in the lungs
of amphibians, reptiles, birds, and mammals, and they may
be present in some fishes that build bubble nests.
Time The small dimensions of the fragile alveolar sacs
create mechanical problems that might cause them to
Figure 13-34 Ventilation in the frog is a stepwise process. Shown here collapse. To understand why alveolar collapse is a prob-
are pressure and volume changes in the buccal cavity and lung of a frog
during buccal movements alone with the glottis closed and during buc-
lem, and how surfactants counteract it, consider a tiny
cal and lung movements with the glottis open and the nares closed (i.e., bubble that is alternately inflated and then deflated. As
lung filling). [Adapted from West and Jones, 1975.1 discussed in Chapter 12, Laplace's law states that the
pressure differential between the inside and outside of a
bubble is proportional to 2y/R, where y is the wall
tension per unit length and R the radius of the bubble. If
Invertebrates two bubbles have a similar wall tension but a different
Invertebrates exhibit a variety of gas-transfer mecha- radius, the pressure in the small bubble will be higher
nisms. Ventilation does not occur in some invertebrates, than that in the large bubble. As a result, if the bubbles are
which rely only on diffusion of gases between the lung joined, the small bubble will empty into the large bubble
and the environment. In spiders, which have paired venti- (Figure 13-35A,B).
lated lungs on the abdomen, the respiratory surface con- A somewhat similar situation exists in the lung. We can
sists of a series of thin, blood-filled plates that extend like consider the alveoli as a number of interconnected bubbles.
the leaves of a book into a cavity guarded by an opening If the wall tension is similar in alveoli of different size, the
(spiracle).The spiracle can be opened or closed to regu- small alveoli will tend to collapse, emptying into the larger
late the rate of water loss from these "book lungs." Snails alveoli. This normally does not occur in the lung for two
and slugs have ventilated lungs that are well-vascularized reasons: surrounding tissue helps prevent overexpansion of
invaginations of the body surface, the mantle cavity. The alveoli, and the properties of the alveolar surfactant lining
volume change that the snail lung is capable of undergo- are such that wall tension increases when the surface film is
ing enables the animal to emerge from and withdraw into expanded and decreases when it is compressed. This occurs
its rigid shell. When the snail retracts into its shell, the because the film expands as alveolar volume increases, so
lungs empty, a situation similar to that seen in tortoises. the surfactant spreads out and therefore is less effective in
GAS E X C H A N G E A N D A C I D - B A S E B A L A N C E 547
......................................
lowering surface tension (Figure 13-392).The result of this importance. It is therefore not surprising that these ani-
effect is to minimize pressure differences between large and mals in particular have evolved means of minimizing the
small alveoli, thus reducing the chance of collapse. Alveoli loss of water.
fold as their volume decreases, and the regions between the The rates of heat and water loss from the lung are inti-
folds come to have a thick layer of surfactant. The very low mately related. As air is inhaled, it is warmed and humidi-
surface tension of this thick surfactant layer permits easy fied by evaporation of water from the nasal mucosa. Be-
inflation of collapsed and folded alveoli. If only water were cause the evaporation of water cools the nasal mucosa, a
present in the folds, large forces would be required to sep- temperature gradient exists along the nasal passages. The
arate the layers and inflate the alveoli. nose is cool at the tip, increasing in temperature toward the
In mammals, surfactants appear in the fetal lung prior glottis. As the moist air leaving the lung is cooled, water .
to birth, thereby reducing the forces required to inflate the condenses on the nasal mucosa, since the water vapor pres-
lungs of the newborn. Newborns who produce no lung sur- sure for 100% saturation decreases with temperature. Thus
factants cannot inflate their lungs at birth without assis- the cooling of exhalant air in the nasal passages results in
tance. This condition, referred to as newborn respiratory the conservation of both heat and water. The blood circu-
distress syndrome, occurs primarily in premature babies. lation to the nasal mucosa is capable of supplying water to
Assistance can be given to the baby by forcing air into the saturate the inhalant air, but the temperature gradients
lungs, using positive pressure ventilation, and by surfactant established by water evaporation and air movement are not
replacement. In addition, pregnant women who are likely destroyed by the circulation.
to have a premature birth can be given an injection of cor- The structure of the nasal passages in vertebrates is
tisol during gestation to stimulate surfactant production in variable, and to some extent it can be correlated with the
the fetus. ability of animals to regulate heat and water loss. Humans
have only a limited ability to cool exhaled air, which is sat-
Heat and Water Loss across the Lung urated with water vapor and is at a temperature only a few
Increases in lung ventilation not only increase gas transfer degrees below core body temperature. Other animals have
but also result in more loss of heat and water. Thus, the longer and narrower nasal passages for more effective wa-
evolution of lungs has involved some compromises. Air in . ter conservation, as we will discuss in Chapter 14.
contact with the respiratory surface becomes saturated Poikilotherms such as reptiles and amphibians, whose
with water vapor and comes into thermal equilibrium body temperatures adjust to the ambient temperature, ex-
with the blood. Cool, dry air entering the lung of mam- hale air saturated with water at temperatures about
mals is humidified and heated. Exhalation of this hot, hu- 0.5- l.O°C below body temperature. Pulmonary air tem-
mid air results in considerable loss of heat and water, peratures and body surface temperatures are often slightly
which will be proportional to the rate of ventilation of the below ambient because of the continual'evaporation of wa-
lung surface. Many air-breathing animals live in very dry ter. In some reptiles, however, body temperature is main-
environments, where water conservation is of paramount tained above ambient. In the iguana, heat and water loss is
Laplace's law
I
Decreas~ngth~ckness
of surfactant film
Figure 13-35 The presence of surfactant in the lungs helps prevent alve- the large bubble with the lower pressure (C)The tendency of small alve-
olar collapse. (A) Laplace's law states that the pressure (P) in a bubble de- 011to collapse Into larger alveoli In the lung IS amellorated by a surfactant
'
creases with increased radius (R) if the wall tension (yj remains constant. l ~ n ~ nAs
g the surfactant film expands w ~ t hthe alveolus, the th~cknessof
Thus, if two bubbles have the same wall tension but the radius of one is the film decreases and the surface tens~onIncreases Because the surface
twice that of the other, the pressure in the small bubble is two times that tens~onis a major component of the wall tenslon, th~seffect tends to mln-
of the large bubble. The equation is written 4ylR rather than 2ylR be- lmlze pressure differences between alveol~of different slzes, thereby sta-
cause the bubble in air has an inner and an outer surface. (B) If the bub- b~lrz~ng them
bles are joined, the small bubble with the higher pressure collapses into
548 I N T E G R A T I O N O F PHYSIOLOGICAL SYSTEMS
.........................................
controlled in a manner similar to that observed in mammals. flow and volume, an increase in hematocrit and blood oxy-
In addition, this lizard conserves water by humidifying air gen affinity, and an increase in the Po> difference across the
with water evaporated from the excretory fluid of the nasal eggshell (Figure 13-36B).The eggshell, once produced, does
salt glands. The rate of water loss is closely correlated with not change during the development of the embryo.
lung ventilation and, therefore, oxygen uptake. Reptiles gen- Water is lost from the egg during development, causing
erally have much lower oxygen requirements than mammals a gradual enlargement of an air space within the egg. The
or birds, and so their rate of water loss is much less. volume of this air cell is as much as 12 ml at hatching in the
chicken egg. Just before they hatch, birds ventilate their
Gas Transfer in Bird Eggs lungs by poking their beaks into the air cell. Blood PCO2 is
The shells of bird eggs have fixed dimensions but contain an initially low in the embryo, but it gradually rises to about
embryo whose gas-transfer requirements increase by a fac- 45 mm Hg just before hatching (Figure 13-36C). This pres-
tor of lo3between laying and hatching. Thus, the transfer sure is maintained after hatching, thus avoiding any
of 0, and CO, must take place across the shell at ever-in- marked acid-base changes when the bird switches from the
creasing rates during development while the dimensions of shell to its lungs for gas exchange.
the transfer surface (eggshell)do not change. Gases diffuse The shell and underlying membranes, representing the
through smabl air-filled pores in the eggshell and then barrier between ambient air and embryonic blood, can be
through underlying membranes, including the chorioallan- divided into an outer gas phase (the air cell) and an inner
toic membrane (Figure 13-36A).The chorioallantoiccircu- liquid phase. At sea level the outer gas phase represents
lation is in close apposition to the eggshell and increases about 30%-40% of the total diffusive resistance to oxy-
with the development of the embryo. Several factors con- gen transfer, 85% of that to carbon dioxide, and 100% of
tribute to the increase in gas-transfer rates during develop- that to water vapor. Eggs at altitude are exposed to a re-
ment in the bird's egg: development of an underlying circu- duction in both oxygen and total gas pressure. The rate of
lation in the chorioallantoic membrane, an increase in blood diffusion of gases increases with a reduction in total pres-
Allantoic
venous blood
(oxygenated)
.
insect5 tracheal system, and between the
- . - - .- . -.
.
- - . - . ... bubble and water. The direction of gas flow
. - - ~. -
will depend on the partial pressures of 0,,
Water: CO,, and N, and the total pressure (P) in bub-
P o p = 150mmHg bles under water. (A) Conditions at start of de-
PNp= 592.7 mrn Hg scent. (B) Condition in bubble immediately af-
ter being taken to depth of 1 m. (C)Condition
sometime later at same depth. Arrows indi-
cate diffusion of gas molecules. Note that the
sum of the gas partial pressures In the water
phase (and in the atmosphere) always equals
Po2 = 150mrnHg
PNz= 592 7 mm Hg
Water =
"02
150 mm ~g =
2~'
593 mm Hg f Water
P = 743 mm Hg
Air
P =
Air
Cuticle
Tissue
Tracheal
system
Figure 13-40 Hydrofuge hairs on the surface of some insects and insect and then into the animal via the tracheal system. Typically, there are about
eggs have an incompressible air space that acts as a gill under water. 106hairs per mm2; only a few are depicted here. (B) Partial pressures of
(A) Schematic diagram of plastron with protruding hydrofuge hairs. Oxy- oxygen and nitrogen in the air and water phases.
gen diffuses from water into the air space contained within the plastron
Figure 13-43 Various arrangements for the flows of water and blood at
the respiratory surface are found in aquatic animals. Relative changes in
Po, in water and blood are indicated below each diagram. I, inhalant; E,
exhalant; a, arterial blood; v, venous blood.
I Filament
Blood
c u t across
vessel
Septum
Lamella
Blood
flow canal
Figure 13-45 The general structure of g~llsis similar in all fishes, although rnent with three secondary folds (lamellae) on each side. The flow of
minor variat~onsare found among species. (A) Position of the four gill blood (red arrows) is in the opposite direction to that of the water (black
arches beneath the operculum on the left side of a teleost fish. (B) En- arrows). (D) Part of the dogfish gill. As in teleost fish, the flow of blood is
larged view of part of two gill arches showing the filaments of adjacent in the opposite direction to that of the water. [Parts A-C adapted from
rows touching at their tips. Also shown are the blood vessels that carry the Hughes, 1964; part D adapted from Grigg, 1970.1
blood before and after its passage over the gills. (C) Part of a single fila-
or branchial cavity for this purpose (seeAir-BreathingFishes movement of 0, and CO, between the environment and
in Chapter 12).Air-breathing crabs usually show a decrease mitochondria in mammals is regulated by altering lung
in oxygen consumption as well as a decrease in body carbon ventilation and the flow and distribution of blood within
dioxide levels when they move from air to water breathing. the body. Here we place emphasis on the control of breath-
The purple shore crab, Leptograspus variegatus, however, ing; Chapter 12 presents details of the control of the car-
shows no change in body oxygen content as it moves be- diovascular system.
tween air and water and may regulate body carbon dioxide
and therefore pH levels by adjusting the ratio of air to wa- Ventilation-to-Perfusion Ratios
ter breathing. Thus this crab truly is amphibious. Energy is expended in ventilating the respiratory surface
with air or water and in perfusing the respiratory epithe-
lium with blood. The total cost of these two processes is dif-
REGULATION OF GAS TRANSFER ficult to assess, but probably amounts to 4%- 10%of the
AND RESPIRATION total aerobic energy output of an animal, depending on the
Because the regulation of the rate of 0, and C 0 2 transfer species in question and the physiological state of the ani-
has been studied most extensively in mammals, this section mal. Thus, gas transfer between the environment and cell
focuses on mammalian regulation of gas transfer. The accounts for a considerable proportion of the total energy
556 I N T E G R A T I O N O F PHYSIOLOGICAL SYSTEMS
........................................
A Secondary
I , lamellae
Water
Mucus
Epithelial
cell
Pillar cell
Basement
---
Water flow
Blood flow 50 p m
membrane
Blood plasma
Figure 13-46 Water flows between gill lamellae, which are covered by a verse sertion through a trout gill lamella showing components of the
thin ep~theliallayer. (A) Scanning electron micrograph of a plastic cast of water-blood barrier. [Courtesy of 6. J. Gannon.]
thevasculatureof a trout gill filament, showing several lamellae. (B)Trans-
output of the animal and represents a significant selective The ventilation-to-perfusion ratio must be maintained
pressure in favor of the evolution of mechanisms for the over each portion of the respiratory surface as well as over
close regulation of ventilation and perfusion in order to the whole surface. The pattern of capillary blood flow can
conserve energy. change in both gills and lungs, changing the distribution of
The rate of blood perfusion of the respiratory surface is blood over the respiratory surface. The distribution of air
related to the requirements of the tissues for gas transfer or water must reflect the blood distribution. Perfusion of
and to the gas-transport capacities of the blood. To ensure an alveolus without ventilation is as pointless as ventilat-
that sufficient oxygen is delivered to the respiratory surface ing an alveolus without blood perfusion of that same alve-
to saturate the blood with oxygen, the rate of ventilation, olus. Although such extreme situations are unlikely to oc-
v*, must be adjusted in accord with the perfusion rate, cur, the maintenance of too high or too low a blood flow
Q, and the gas content of the two media so that the amount or ventilation rate will result in energetically inefficient gas
of oxygen delivered to respiratory surface equals that taken transfer per unit of energy expended. For efficient gas
away in the blood. The oxygen content of arterial blood in transfer, the optimal ventilation-to-perfusion ratio should
humans normally is similar to that of air. The %lQ ratio, be maintained over the whole respiratory surface. This op-
therefore, is about 1 in humans (Figure 13-47A). Water, timal maintenance does not preclude differential rates of
however, contains only about one-thirtieth as much dis- blood perfusion over the respiratory surface, but requires
solved oxygen as an equivalent volume of air at the same only that the flows of blood and inhalant medium be
Pol and temperature. Thus, in fishes, the ratio of water flow, matched.
V,, over and blood flow, Q , through the gills is between The efficiency of gas exchange is diminished if some of
10 : 1 and 20 :1 (Figure 13-47B), much higher than the li, the blood entering the lungs or gills either bypasses the res-
/Q ratio in air-breathing mammals. Based on the difference piratory surface or perfuses a portion of the respiratory sur-
in the oxygen content of water and air, the VG/Qratio in face that is inadequately ventilated (Figure 13-48). The
fishes might be expected to be 30: 1. However, it is lower magnitude of such venous shunts, expressed as a percent-
than this because the oxygen capacity of the blood of lower age of total flow to the respiratory epithelium, can be cal-
vertebrates is often only half that of mammalian blood. culated from the arterial and venous 0, content, assuming
Any changes in the oxygen content of the inhalant an ideal arterial 0, content. In the lung, for instance, blood
medium will affect the VA/Qratio. In order to maintain a is almost in equilibrium with alveolar gas tensions. If these
given rate of oxygen uptake, a decrease in Po2of inhalant tensions and the blood oxygen dissociation curves are
air or water must be compensated for by an increase in known, the expected ideal 0, content of arterial blood
ventilation and hence an increase in the ventilation-to- can be determined. Let us assume that this ideal content
perfusion ratio. Conversely, an increase in the inhalant Pol is 20 ml of 0, per 100 ml of blood (20 vol %) and the
is accompanied by a decrease in ventilation if the rate measured values for arterial and venous blood are 1 7 and
of oxygen uptake remains the same. 5 vol %, respectively. This reduction in measured arterial
GAS EXCHANGE A N D ACID-BASE BALANCE 557
...............................................................................
A Lung (human) Figure 13-47 The ventilation-to-perfusion ra-
Lung ventilation = 7500 rnl. min-I tio in fish gills is much higher than in the hu-
man lung. Approximations of volumes and
Breathing rate = I$ \ .
Alveolar ventilat~on(V,) = 5200 ml min-I flows in the human lung and trout gill are
shown; actual values may vary considerably.
15 min-I
Volumes:
Alveolar gas = 3000
Pulmonary capillary
blood = 70 ml
Blood
.
Flow ( t , ) = 40 rnl min-'
.
Flow (Q) = 4 ml min-'
r \;,I& 10
-
L 0, content from the ideal situation can be explained through one or more venous shunts. This is an extreme ex-
- '
in terms of a venous shunt, oxygenated arterial blood ample to illustrate a point; in most cases, venous shunts are
b (20~ 0 1 %being
) mixed with venous blood ( 5~ 0 1 %in) the very small.
ratio of 4 : 1to give a final arterial 0, content of 17 ~ 0 1 % ; Flows of blood and inhalant medium (air or water) are
that is, 20% of the blood perfusing the lung is passing regulated to maintain a near optimal ventilation-to-perfu-
Gill
No
water
flow
i Lung
Gill
Figure 13-48 The efflc~encyof gas transfer In the lung and g~llsIS de- enough to the resp~ratory
eplthel~um(shunt 2) Bloodflow IS regulatedto
creased when blood flows to a portlon of the respiratory surface without avo~dthe development of such venous shunts In the lung and gills
adequate vent~lat~on (shunt 1) or because blood does not flow close
558 I N T E G R A T I O N O F P H Y S I O L O G I C A L SYSTEMS
.........................................
sion ratio over the surface of the respiratory epithelium un- which have a reflex inhibitory effect, via the vagus nerve, on
der a variety of conditions. In general terms, Q is regulated the medullary inspiratory center (nucleus tractus solitarius)
to meet the requirements of the tissues; .ir,and vGare reg- and therefore on inspiration. Thus the medulla contains a
ulated to maintain adequate rates of 0, and CO, transfer. central rhythm generator that drives the pattern generator
Such mechanisms as hypoxic vasoconstriction of blood ves- within the medullary respiratory center to cause breathing
sels help to maintain optimal ventilation-to-perfusion movements. This system is modified by inputs from other
ratios in different parts of the respiratory surface. As dis- areas of the brain and from various peripheral receptors.
cussed earlier, low alveolar oxygen levels cause a vaso- The medullary respiratory center contains inspiratory
constriction in lung vessels, thereby reducing blood flow to neurons, whose activity coincides with inspiration, and
poorly ventilated, and therefore hypoxic, regions and in- expiratory neurons, whose activity coincides with expira-
creasing blood flow to well-ventilated regions of the respi- tion. The respiratory rhythm was once considered to arise
ratory surface. Blood perfusion of the respiratory surface from reciprocal inhibition between inspiratory and expira-
tends to be less well distributed in resting animals. Blood tory neurons, with reexcitation and accommodation oc-
pressure rises with exercise and blood is distributed more curring within each set of neurons. But several lines of evi-
evenly under these conditions, resulting in a more even dence indicate this model of the central rhythm generator
ventilation-to-perfusion ratio over the respiratory surface. is not tenable, and more recent studies suggest that respi-
ratory rhythm depends primarily on the activity of inspira-
Neural Regulation of Breathing tory neurons.
The integration of breathing movements in all air-breath- Inspiratory neuronal activity, recorded from either the
ing vertebrates results from the central processing of many phrenic nerve or some individual neurons in the medulla,
sensory inputs. The central processor consists of a pattern shows a rapid onset, a gradual increase, and then a sharp
generator, determining the depth and amplitude of each cutoff with each burst of activity associated with in-
breath, and a rhythm generator, controlling breathing fre- halation. This neuronal activity results in a contraction of
quency. Several sensory inputs adjust ventilation to main- the inspiratory muscles and a decrease in intrapulmonary
tain adequate rates of gas transfer and blood pH. Other in- pressure (Figure 13-49A).Increased blood CO, levels cause
puts integrate breathing movements with feeding, talking the progressive growth of inspiratory activity to increase
and singing, or other body movements. Certain sensory in- more rapidly (Figure 13-49B). Thus, the rate of rise of in-
puts may cause coughing or swallowing reflexes, which spiratory activity is increased by inputs from chemorecep-
protect the respiratory epithelium from environmental haz- tors, resulting in a more powerful inspiratory phase. The
ards. Other inputs function to optimize breathing patterns "off switch" of inspiratory neurons occurs once activity
to minimize energy expenditure. in the neuron has reached a threshold level. Expansion of
the lung stimulates pulmonary stretch receptors, whose ac-
Medullary respiratory centers tivity reduces the threshold for the inspiratory off switch
As noted earlier, the mammalian lung is ventilated by the (Figure 13-49C). Thus the pulmonary stretch receptors,
action of the diaphragm and muscles between the ribs (see through their action on inspiratory neurons, prevent over-
Figures 13-29 and 13-31). These muscles are activated expansion of the lung.
by spinal motor neurons and the phrenic nerve, which re- The interval between breaths is determined by the in-
ceive inputs from groups of neurons that constitute the terval between bursts of inspiratory neuronal activity,
medullary respiratory centers. The control of respiratory which is related to the level of activity in the previous burst
muscles can be very precise, allowing extremely fine control and in afferent nerves from pulmonary stretch receptors. In
of air flow, as is required for such complex actions in hu- general, the greater the level of inspiratory activity (i.e., the
mans as singing, whistling, and talking, as well as simply deeper the breath), the longer the pause between inspira-
breathing. Microsections of the neonatal rat brain stem tions. The result is that the ratio of inspiratory to expira-
indicate that the ere-Botzinger complex in the ventral tory duration remains constant in spite of changes in the
medulla is capable of generating the respiratory rhythm length of each breathing cycle. This ratio is affected by the
and may represent the central rhythm generator that main- level of activity of the pulmonary stretch receptors. If, for
tains breathing rhythm in the adult. Rhythmic activity is example, the lung empties only slowly during expiration,
enhanced by neurons in the pons and medulla, and some the pulmonary stretch receptors will remain active while
neurons just anterior to the medulla cause prolonged in- the lung remains inflated; the continued activity of the
spiration in the absence of rhythmic drive from the pons. stretch receptors will prolong the duration of expiration
In 1868 Ewald Hering and Josef Breuer observed that and increase the time available for exhalation. The neu-
inflation of the lungs decreases the frequency of breathing. ronal mechanisms causing phasic activation of inspiratory
(Breuer later became an early proponent of psychoanalysis neurons are poorly understood, as is the nature of the cen-
and collaborated with Sigmund Freud in producing a book tral rhythm generator, possibly located in the pre-Botzinger
on hysteria.) The Hering-Breuer reflex is abolished by cut- complex in the ventral medullary region of the brain.
ting the vagus nerve. Inflation of the lung stimulates pul- Exhalation often is a largely passive process, which
monary stretch receptors in the bronchi and/or bronchioles, does not depend on activity in expiratory neurons. This is
GAS E X C H A N G E A N D A C I D - B A S E B A L A N C E 559
...............................................................................
lntrapulmonary pressure
Decreasing
pressure
lnsplration
3rnrnHg L
200 rns
t
Activity f r o m
lncreasing pulmonary stretch
activity
receptor
Increasing
activity
/
activity
activity ' u
Figure 13-49 Phrenic nerve activity, stimulated by increasing alveolar P,C02, the more rapid the rise of phrenic nerve adivity during inspira-
Pco,, induces insp~rat~on. (A) Relationship between phrenic nerve activ- tion. (C) Effect of increasing activity from pulmonary stretch receptors o n
ity and intrapulmonary pressure during inspiration. Note the sudden on- activity in the phrenic nerve. In the absence of stretch receptor activity,
set, gradual rise, and then "off switch," or termination, of inspiratory ac- the switch off in phrenic nerve activity is delayed (red traces). An increase
tivity (B)Effect of increasing alveolar Pco, levels (P,CO,) on discharge in in receptor activity results in an earlier termination of activity in the
the phrenic nerve. Recordings were made at P,C02 ranging from phrenic nerve but does not affect the rate of increase in phrenic activity
28.5 mm H g (bottom trace) to 60 m m Hg (top trace). The higher the before the switching off (black traces).
especially true during quiet, normal breathing. Expiratory blood, reducing peripheral chemoreceptor input to the res-
neurons are active only when inspiratory neurons are qui- piratory center. Breathing stops until oxygen levels fall suf-
escent, and then they show a burst pattern somewhat sim- ficiently to increase chemoreceptor drive enough to initiate
ilar to, but out of phase with, that of inspiratory neurons. breathing again. This results in the periodic breathing typ-
Inspiratory neuronal activity inhibits expiratory activity, ical of many sleeping mammals. In the awake mammal cen-
showing the dominance of inspiratory neurons in the gen- tral respiratory drive is sufficient to maintain continuous
eration of rhythmic breathing. In the absence of inspiratory rhythmic breathing.
activity, expiratory neurons are continually active. Inspira-
tory neuronal activity, however, imposes a rhythm, via in- Factors affecting the rate and depth of breathing
hibition, on expiratory neurons. Several types of receptors respond to stimuli that influence
Fish, birds, and awake mammals usually breath rhyth- ventilation, causing reflex changes in the rate and/or depth
mically and continuously, whereas amphibians and reptiles of breathing. Among the stimuli affecting ventilation are
often show episodic breathing, with pauses between changes in O,, CO,, and pH; emotions; sleep; lung infla-
episodes of rhythmic breathing. Recent studies of the bull- tion and deflation; lung irritation; variations in light and
frog brain stem have shown that these episodic patterns of temperature; and the requirements for speech. These influ-
breathing are an intrinsic property of the brain stem and do ences are integrated by the medullary respiratory centers.
not depend on sensory feedback. The nucleus isthmi in the Breathing can also, of course, be controlled by conscious
bullfrog brain stem is involved not only in the integration volition.
of chemoreceptor input but also appears to be essential for In most, if not all, animals, changes in 0, and CO, lead
the maintenance of episodic breathing. In sleeping mam- to reflex changes in ventilation. The chemoreceptors in-
mals episodic breathing appears to be the result of the in- volved have been localized in only a few groups of animals.
teraction between peripheral and central components of the Chemoreceptors monitor changes in 0, and CO, in arter-
control system. During sleep, central respiratory drive is re- ial blood in the carotid bodies and aortic bodies of mam-
duced in mammals, and breathing is maintained by input mals, in the carotid body of birds, and in the carotid
from peripheral chemoreceptors. A breathing period in- labyrinth of amphibians. In teleost fish chemoreceptors lo-
creases oxygen and decreases carbon dioxide levels in the cated in the gills respond to reductions in O2levels in the
560 I N T E G R A T I O N O F PHYSIOLOGICAL SYSTEMS
.......................................
water and the blood. In all cases, the chemoreceptors are by sustentacular (type 11) cells. The glomus cells, thought
innervated by branches,of the ninth (glossopharyngeal)or to be the actual receptors, are small ovoid cells with a large
tenth (vagus)cranial nerve. nucleus and many dense-core vesicles, or granules (Figure
Mammals and probably other air-breathingvertebrates 13-SOB).These cells are interconnected by synapses and
also have central chemoreceptors, located in the medulla, often possess cytoplasmic processes of different lengths.
that drive ventilation in response to decreases in the pH of They are innervated by afferent fibers of the glossopha-
the cerebrospinal fluid (CSF),usually caused by elevations ryngeal nerve and possibly preganglionic sympathetic ef-
in Pco,. Stimulation of this system is required to maintain ferent~.A single nerve fiber may innervate 10 or 20 glomus
normal breathing: if body PCol falls, or is held at a low level cells. A glomus cell may be either presynaptic or postsy-
experimentally, breathing will cease. These central naptic, or both (reciprocal),with respect to a nerve fiber. A
chemoreceptors have little ability to respond to falling O2 single nerve fiber may be postsynaptic (afferent)to one glo-
levels; the peripheral chemoreceptors have this role and mus cell with a presynaptic connection (efferent) to a
are important in increasing ventilation during periods of neighboring glomus cell or even another region of the
hypoxia. same glomus cell. Many glomus cells lack innervation but
The carotid and aortic bodies of mammals receive a are synaptically connected to other glomus cells in the lob-
generous blood supply and have a high oxygen uptake per ule. A few glomus cells may be innervated by sympathetic
unit weight (Figure 13-50A). These arterial chemorecep- efferent fibers.
tors consist of a number of lobules, or "glomoids," that The chemoreceptors in the carotid and aortic bodies are
surround very convoluted capillaries. The blood vessels stimulated by decreases in blood 0, and pH and increases
can be divided into small and large capillaries and arteri- in blood CO, .It is possible that the observed response to in-
ovenous shunts. The arterioles are innervated by both creasing CO, is due to changes in pH within these receptors
sympathetic and parasympathetic postganglionic efferents. rather than to changes in CO, per se. The result of chemore-
Each lobule consists of several glomus (type I) cells covered ceptor stimulation is to recruit new fibers and increase the
Carotid body
chemoreceptors
/
'Aortic
bodies
Figure 13-50 In mammals, chemoreceptors in the carotid and aortic mus cells. These are connected by synapses and innervated by glos-
bodies monitor blood gas levels and pti. (A) Diagram showing the loca- sopharyngeal afferentfibers. Some regions of afferent nerve endings are
tion of carotid and aortic body chernoreceptors and carotid sinus and presynaptic to the glomus cell, some postsynaptic, and some form
aortic arch baroreceptors (small red dots) in the dog. The baroreceptors reciprocal synapses. A , presynaptic regions. [Part A adapted from
help regulate arterial blood pressure (see Chapter 12). (B) Small portion Comroe, 1962; part B adapted from M.cDonald and Mitchell, 1975.1
of the rat carotid body, which consists of several lobules containing glo-
firing rate in the afferent nerves innervating glomus cells.
The chemoreceptors .adapt to changing arterial CO, levels.
The carotid body chemoreceptors show a much larger re-
sponse to pH and/or C 0 2 changes than the aortic body
chernoreceptors. Stimulation of these chemoreceptors leads
to an increase in lung ventilation, mediated via the
medullary respiratory center. The actual increase, in re-
sponse to a given decrease in arterial Pol, depends on the
blood CO, level, and vice versa (Figure 13-51). Efferent ac-
tivity to the carotid body modulates the response. Increased
sympathetic efferent activity constricts arterioles in the
carotid body via an a-adrenergic mechanism, thereby re-
ducing blood flow, which in turn increases the chemorecep-
tor discharge and lung ventilation. Nonsympathetic efferent
activity in the carotid nerve reduces the response of the
carotid body to changes in arterial blood Pol and Pco, Figure 13-52 Central H+-sensitivereceptors are influenced by the pH of
cerebrospinalfluid (CSF) and by arterial Pco, Carbon dioxide molecules
and/or pH. Increases in temperature and osmolarity also diffuse readily across the walls of the brain capillaries and alter CSF pH,
stimulate the arterial chemoreceptors, and stimulation of the but there is a barrier to other molecules.An increase in Pco2causes a de-
carotid nerve causes increased ADH release. Thus the crease in the CSF pH; the resulting stimulation of H+ receptors reflexively
carotid body chemoreceptors may play a role in osmoreg- increases breathing.Across some capillarywalls, exchange of HCO, and
ulation as well as in the control of breathing and circulation. C I helps to maintain a constant pH of the CSF in the face of a prolonged
change in PC02.
As mentioned previously, mammals and possibly other
air-breathing vertebrates have central chemoreceptors that In mammals and other air-breathing vertebrates, car-
are necessary for normal breathing. These H+-sensitive re- bon dioxide rather than oxygen levels dominate in the
ceptors are located in the region of the medullary respira- control of breathing. In aquatic vertebrates, however, oxy-
tory center and are stimulated by a decrease in the pH of gen is the major factor in the control of breathing. In fact,
the CSF. The CSF of mammals, and possibly of other ver- fishes exposed to high oxygen levels will reduce breathing
tebrates, is very low in protein and is essentially a solution to the extent that there is a marked increase in Pco2in the
of NaCl and NaHCO,, with low but closely regulated lev- blood. Two factors account for this difference. First, oxy-
els of K+, Mg2+,and Ca2+.The CSF is also poorly buffered; gen concentration is much more variable in the aquatic
therefore small changes in PCo2have a marked effect on environment than in air. Second, oxygen is much less sol-
CSF pH. Because the blood-brain barrier is relatively im- uble than carbon dioxide in water; as a result, if ventila-
permeable to H+,the central H+-sensitive chemoreceptors tion is adequate to deliver oxygen to the gills, it will also
are insensitive to changes in blood pH. However, changes be adequate to remove carbon dioxide from the blood.
in blood PCOL cause corresponding changes in the PCo2of Under most conditions ventilation does not limit carbon
the CSF, and these in turn result in changes in the pH of the dioxide excretion in aquatic animals. Only in the rare
CSF. An increase in PC,> leads to a decrease in the pH of the condition of very high oxygen levels in the water is venti-
CSF; subsequent stimulation of the H+-sensitive receptors lation reduced sufficiently to curtail carbon dioxide ex-
causes reflex increases in breathing (Figure 13-52). Pro- cretion in fish.
longed changes in PC,! result in the adjustment of the pH The lungs contain several types of receptors that help
of the CSF by changes in H C 0 3 levels. regulate inflation and prevent irritation of the respiratory
surface. We saw earlier that stimulation of pulmonary
stretch receptors prevents overinflation of the lung (Hering-
Breuer reflex).In mammals increased CO, levels reduce the
inhibitory effects of these ~ u l m o n a stretch
r~ receptors on
the medullary respiratory center, thereby increasing the
depth of breathing and lung ventilation. It is not clear
whether the C02-sensitive receptors found in the lungs of
birds are pure C 0 2 chemoreceptors or C0,-sensitive
mechanoreceptors, as observed in mammals. Increased
CO, in the lungs of birds, however, has a greater effect on
" sensory discharge from the lung than that observed in
120 240 360 mammals.
Arterial oxygen Po2(mm Hg) In addition to pulmonary stretch receptors, a variety of
Figure 13-51 Lung ventilation rates increase with a decrease in arterial
irritant receptors are present in the lung. Stimulation of
PO2and an increase in arterial Pco2.The relationships shown here are from these receptors by mucus and dust or other irritant particles
measurements in the duck. [From Jones and Puwes, 1970.1 causes reflex bronchioconstriction and coughing. A third
group of receptors in the lung is positioned close to the pul- and easily avoided. There is, of course, a gradual reduc-
monary capillaries in interstitial spaces; these are called tion in Pol with altitude, and animals vary in their capac-
juxtapulmonary capillary receptors, or type J receptors. ity to climb to high altitudes and withstand the accompa-
These receptors were previously termed "deflation recep- nying reduction in ambient oxygen levels. The highest
tors," but their natural stimulus appears not to be lung de- permanent human habitation is at about 5800 m, where
flation but an increase in interstitial volume, as seen, for ex- the Po, is 80 mm Hg compared to about 155 mm Hg at
ample, during pulmonary edema. Stimulation of type J sea level. Many birds migrate over long distances at alti-
receptors elicits a sensation of breathlessness. Violent exer- tudes above 6000 m, where atmospheric pressures would
cise probably results in a rise in pulmonary capillary pres- cause severe respiratory distress in many mammals. High
sure and an increase in interstitial voluine, which could altitudes are associated with low temperatures as well as
cause stimulation of type J receptors and therefore breath- low pressures, and this also has a marked effect on animal
lessness. distribution.
A reduction in the Po, of ambient air results in a de-
crease in blood P,.,-' which in turn stimulates the carotid and
RESPIRATORY RESPONSES TO
aortic body chemoreceptors, causing an increase in lung
EXTREME CONDITIONS
ventilation in mammals. The rise in lung ventilation then
Variations in the levels of respiratory gases, diving by air- leads to an increase in CO, elimination and a decrease in
breathing animals, and exercise all induce respiratory blood PCo2.The decrease in blood PCoLcauses a reduction
responses. Let's see how animals adjust to these extreme in Pco, and therefore an increase in p H of the CSF. De-
conditions. creases in blood PC,> and increases in CSF p H tend to re-
duce ventilation, thereby attenuating the hypoxia-induced
Reduced Oxygen Levels (Hypoxia) increase in lung ventilation. If, however, hypoxic conditions
Aquatic animals are subjected t o more frequent and are maintained, as occurs when animals move to high alti-
rapid changes in oxygen levels than are air-breathing an- tude, both blood and CSF pH are returned to normal lev-
imals. Both mixing and diffusion are more rapid in air els by the excretion of bicarbonate. This process takes about
compared with water, so regions of local hypoxia develop one week in humans. Thus, as CSF p H returns to normal,
more often in aquatic environments. Although photosyn- the reflex effects of hypoxia on ventilation predominate; the
thesis can cause very high oxygen levels during the day in result is a gradual increase in ventilation as the animal ac-
some aquatic environments, oxygen consumption by both climatizes to altitude. The response to prolonged hypoxia
biological and chemical processes can produce localized may also involve modulation of the effects of CO, on the
hypoxic regions. The changes in oxygen levels in water carotid and aortic bodies to reset these chemoreceptors to
may or may not be accompanied by changes in carbon the new lower CO, level at high altitude.
dioxide.
Many aquatic animals can withstand very long periods
of hypoxia. Some fishes (e.g., carp) overwinter in the bot-
tom mud of lakes where the Pol is very low. Many inverte-
brates also bury themselves in mud with a low Pol but high
nutritive content. Some parasites live in hypoxic regions,
such as the gut, during one or more phases of their life cy-
cle. Limpets and bivalve mollusks close their shells during
exposure at low tide to avoid desiccation, but as a conse-
quence are subject to a period of hypoxia. Many of these As mentioned earlier, low oxygen levels cause a local
animals utilize a variety of anaerobic metabolic pathways vasoconstriction in the pulmonary capillaries in mammals,
to survive the period of reduced oxygen availability. Others producing a rise in pulmonary arterial blood pressure. This
also adjust the respiratory and cardiovascular systems to response normally has some importance in redistributing
maintain oxygen delivery in the face of reduced oxygen blood away from poorly ventilated, and therefore hypoxic,
availability. For instance, aquatic hypoxia causes an in- portions of the lung. When animals are subjected to a gen-
crease in gill ventilation in many fish, as a result of stimu- eral hypoxic environment, however, the increase in the re-
lation of chemoreceptors on the gills. The increase in water sistance to flow through the whole lung can have detri-
flow offsets the reduction in oxygen content and maintains mental effects. Some mammals that live at high altitudes
delivery of oxygen to the fish. In fishes, such as tuna, that exhibit a reduced local pulmonary vasoconstriction in re-
ram ventilate their gills by swimming forward with their sponse to hypoxia; this is probably a genetically determined
mouth open, the size of the gap increases with hypoxia to acclimation. Humans residing at high altitudes are usually
increase water flow over the gills. small and barrel chested, and have large lung volumes.
Compared with aquatic environments, oxygen and Lung development is oxygen insensitive, but the growth of
carbon dioxide levels are relatively stable in air, and local limbs is reduced under hypoxic conditions. The h ~ g hlung
regions of low oxygen or high carbon dioxide are rare to body ratio enables these people to maintain oxygen up-
GAS EXCHANGE A N D ACID-BASE BALANCE 563
.......................................
take under hypoxic conditions. Pulmonary blood pressures Diving by Air-Breathing Animals
are high, and there is often hypertrophy of the right ventri- Many air-breathing vertebrates live in water and dive
cle. High pulmonary pressures produce more even distri- for varying periods of time. Dolphins and whales rise to
bution of blood in the lung, and so augment the diffusing the surface to breathe but spend most of their life sub-
capacity for oxygen. merged. The time between breaths varies with the diver, but
Long-term adaptations also occur during prolonged ex- is around 10-20 minutes for many diving vertebrates
posure to hypoxia. Most vertebrates respond by increasing (Table 13-2).The elephant seal dives regularly to depths of
the number of red blood cells and the blood hemoglobin 400 m, subjecting itself to a pressure of over 40 atm at the
content-and therefore the oxygen capacity of the blood. bottom of the dive. These pressures would crush the tho-
A reduction in blood oxygen levels stimulates production racic cage of man. There are reports of sperm whales div-
of the hormone erythropoietin in the kidney and liver. ing to nearly 2000 m, and staying submerged for over an
Erythropoietin acts on the bone marrow to increase pro- hour. These are of course maximum estimates; most dives
duction of red blood cells (erythropoiesis). Under hypoxic are much shorter and to less depth.
conditions, the levels of hemoglobin-binding organophos- Diving mammals and birds are, of course, subjected to
phates (e.g., DPG) changes, thus altering the oxygen affin- periods of hypoxia during submergence. The mammalian
ity of hemoglobin. In humans, a climb to high altitude is ac- entral nervous system (CNS)cannot withstand anoxia and
companied by an increase in DPG levels and a reduction in must be supplied with oxygen throughout the dive. Diving
the hemoglobin-oxygen affinity. The increasing DPG levels animals solve the problem by utilizing oxygen stores in the
offset the effects of high blood pH on hemoglobin-oxygen lungs, blood, and tissues (Figure 13-53). Many diving ani-
affinity. The high blood pH results from hyperventilation in mals have high hemoglobin and myoglobin levels, and their
response to the low 0, availability. total oxygen stores generally are larger than those in non-
Hypoxia due to travel to high altitude also results in diving animals. To minimize depletion of available stores,
systemic vasodilation and an increase in cardiac output. oxygen is preferentially delivered to the brain and the heart
The higher cardiac output lasts only a few days and returns during a dive; blood flow to other organs may be reduced,
to normal or drops below normal as O2 supplies to tissues and these tissues may adopt anaerobic metabolic pathways.
are restored by the compensatory increases in ventilation There is a marked slowing of heart rate (bradycardia) and
and blood hemoglobin levels. Exposure to hypoxia stimu- a reduction of cardiac output during a prolonged dive or
lates a proliferation of capillaries in tissues, ensuring a more if the animal is forcibly submerged in an experimental set-
adequate oxygen delivery to the tissues. The gills of fishes ting (see Figure 12-47).Air-breathing animals that spend
and amphibians are larger in species exposed to prolonged prolonged periods submerged at sea must have sufficient
periods of hypoxia. Similar enlargement of the respiratory oxygen stores to sustain aerobic metabolism, because they
surface apparently does not occur in mammals. These cannot tolerate the high accumulation of lactic acid that re-
processes augment the transfer of oxygen, its transport in sults from anaerobic metabolism. During prolonged dives,
the blood, and its delivery to the tissues, but they take from metabolic rates and thus oxygen needs often are reduced in
several hours to days or weeks to reach completion. such animals (e.g., elephant seals).
Some diving animals, such as the Weddell seal, exhale
Increased Carbon Dioxide Levels (Hypercapnia)
before diving, thus reducing the oxygen store in their
In many animals, an increase in blood PCo2results in an in-
crease in ventilation. In mammals the increase is propor-
tional to the rise in the CO, level in the blood. The effect is
mediated by modulation of the activity of several receptors TABLE 13-2
Total oxygen stores, mean dive time, and mean dive depth in
that send messages to the medullary respiratory center. diving vertebrates
These receptors include the chemoreceptors of the aortic
Mean dr~ve Mean depth
and carotid bodies and the mechanoreceptors in the lungs, 0, stores tlme of d ~ v e
but the response is dominated by the central H + receptors Spec~es (ml . kg-') (m~nutes) (meters)
(see Figure 13-52).Correction of CSF pH, in the face of al-
Leatherback turtle 20 11 -
tered PCO2levels, is very important in the return of ventila-
tion to normal. Pengu~ns* 58 6 100
A marked increase in ventilation occurs almost im- Weddell seal 60 15 100
mediately in response to a rise in CO, .The increase is main- Norhern elephant - 20 400
seal
tained for long periods in the presence of increased CO,,
but ventilation eventually returns to a level slightly above Humant 20 2 shallow
the volume that prevailed before hypercapnia. This return *The 0, stores are for; King penguin; the dive time and depth are for an
Emperor penguin.
to a value only slightly greater than the initial ventilation
level is related to increases in levels of plasma bicarbonate +Leatherbackturtles have similar oxygen stores as humans but can dive
for much longer times because of their lower rate of oxygen use.
and CSF bicarbonate, with the result that pH returns to nor-
mal even though the raised CO,?levels are maintained. Source: Adapted from Kooyman, 1989.
564 I N T E G R A T I O N O F PHYSIOLOGICAL SYSTEMS
...............................................................................
Exercise
Muscles Exercise increases 0, utilization, C 0 2 production, and
metabolic acid production. Cardiac output increases
to meet the higher demands of the tissues. Even though
lungs the transit time for blood through the lung capillaries
is reduced, nearly complete gas transfer still occurs
(Figure 13-54). Ventilation volume increases in order to
maintain gas tensions in arterial blood in the face of in-
creased blood flow. The increase in ventilation in mammals
is rapid, coinciding with the onset of exercise. This initial
sudden increase in ventilation volume is followed by a more
gradual rise until a steady state is obtained both for venti-
lation volume and oxygen uptake (Figure 13-55).When ex-
ercise is terminated, there is a sudden decrease in breathing,
followed by a gradual decline in ventilation volume. Dur-
ing exercise, 0, levels are reduced and CO, and H+ levels
raised in venous blood, but the mean P02and PCO7 in arte-
rial blood do not vary markedly, except during severe ex-
ercise. The oscillations in arterial blood Pg and PcOlassoci-
ated with each breath increase in magnitude, although the
mean level is unaltered.
r e Air-breath~ng an~malsdraw on cxygen stores In the lungs,
~ i ~ u13-53 Exercise covers a range from slow movements up to
blood, and t~ssues(espec~allymuscles)when submerged The general- maximum exercise capacity. The phrase moderate exercise
zed total oxygen stores (expressed In ml 0,. '
kg body we~ght)of the refers to exercise above resting levels that is aerobic, with
major taxa of marlne d~versare compared w~thhumans The d~str~butlon
only minor energy supplies derived from anaerobic glycol-
of the total 0, stores In the lungs (gray), blood (red), and muscle (whlte)
vanes somewhat among specles [Adapted from Kooyman, 1989 1
ysis. Severe exercise refers to exercise in which oxygen up-
take is maximal and further energy supplies are derived
from anaerobic metabolism. Heavy exercise is a term some-
lungs. During a deep dive, the increase in hydrostatic pres- times used to denote the exercise level between moderate
sure results in lung compression. In those animals that re- and severe exercise.
duce lung volume before a dive, air is forced out of the The onset of exercise involves many changes in lung
alveoli as the lungs collapse and is contained within the ventilation and the cardiovascular system, as well as mus-
trachea and bronchi, which are more rigid but less perme- cle contraction. In the initial stages, during the transition
able to gases. If gases remained in the alveoli, they would from rest to exercise, the animal is not in a steady state and
diffuse into the blood as pressure increased. At the end of part of the energy supply is derived from anaerobic
the dive, the partial pressure of nitrogen in the blood
would be high, and a rapid ascent would result in the for-
mation of bubbles in the blood the equivalent of decom-
pression sickness, or the "bends," in humans. Thus exha-
lation before diving reduces the chances of the bends - -Alveolar
- - - -gas
-----------------
occurring. Since only about 7% of a Weddell seal's total
oxygen stores are in the lungs, pre-dive exhalation appears
to be a reasonable trade-off.
Receptors that detect the presence of water and that in-
hibit inspiration during a dive are situated near the glottis
and near the mouth and nose (depending on the species).
The decrease in blood 0, levels and increase in CO, levels
that occur during a dive do not stimulate ventilation be- m
cause inputs from the chemoreceptors of the carotid and
aortic bodies are ignored by the respiratory neurons while
the animal is submerged.
During birth, a mammal emerges from an aqueous en-
t
61ood
Time in capillar~es
vironment into air and survives a short period of anoxia enters lung
between the time the placental circulation stops and the
Figure 13-54 Blood Po, rapidly reaches near equilibration with alveolar
time air is first The respirator). and re- Po2even during exercise. Although blood flow increases, and therefore
sponses of the fetus during this period are similar in several blood spends less time in the lung capillaries, during exercise, increased
respects to those of a diving mammal. ventilation allows equilibration to occur. [Adapted from West, 1970.1
GAS EXCHANGE A N D ACID-BASE BALANCE 565
......................................
/
Bladder
PO,(atm) from venous to arterial capillaries, the rete acting as a coun-
tercurrent exchanger (see Spotlight 14-2). H. Kobayashi,
B. Pelster, and P. Scheid (1993),howevel; were unable to de-
Water Po, tect any significant transfer of oxygen across the rete. The
Pol does fall in the blood flowing away from the gas gland
because oxygen binds to hemoglobin, not because of any
loss of oxygen to arterial blood entering the rete. Exactly
- 20 40 60 80 100 how and why this occurs will be discussed later.
Depth (m)
Oxygen Secretion
Figure 13-56 The volume of the swimbladder decreases and bladder
Po, increases as a fish descends. Hydrostatic pressure increases by ap- The rete structure reduces gas loss from the swimbladder,
proximately 1 atm every 10 m. In this example, oxygen is assumed to be but how is oxygen secreted into the swimbladder? First, con-
the only gas present and is neitheradded to nor removedfrom the blad- sider the relationship between Po,, oxygen solubility, and
der. Fishes can maintain constant density only by maintaining constant
bladder volume, which is achieved by add~tionof oxygen to the bladder
oxygen content. Oxygen is carried in blood bound to he-
with increasing depth. Note the increasing Po, difference between water moglobin and in physical solution. If oxygen is released from
and bladder with depth. Oxygen must be moved from water into the hemoglobin into physical solution, Pol will increase. The re-
swimbladder agalnst this increasing Po, gradlent. lease of oxygen from hemoglobin can be caused by a reduc-
GAS E X C H A N G E A N D A C I D - B A S E B A L A N C E 567
...............................................................................
To liver
Rete
-
____------------_
To heart
----_ ---.1
Oval
(gas resorbtion)
Esophagus
To l ~ v e r
Duct Swimbladder (gas secretion)
Figure 13-57 Two main types of swimbladder are found in fish. (A) A bladder (e.g.,from the perch, Perca fluviatilis) lacks a dud. Gas enters and
physostome swimbladder (e.g., from the eel, Anguilla vulgaris) is con- leaves the bladder vla the blood. [Adapted from Denton, 1961.I
nected to the outside via a duct to the esophagus. (B) A physoclistswim-
tion in pH via the Root-off shift (Figure 13-58).An increase and two protons for each glucose molecule. The pentose
in ionic concentration reduces oxygen solubility and also re- phosphate shunt, however, is active in the gas gland, pro-
sults in an increase in Po>,as long as the oxygen content in ducing carbon dioxide via decarboxylation of glucose with-
physical solution remains unchanged. Thus, an increase in out oxygen consumption. The production of carbon diox-
blood can be achieved by releasing oxygen from hemo- ide, lactate, and protons by gas-gland cells results in (1)a
globin or increasing the ionic concentration of the blood. decrease in pH, which causes the release of oxygen from he-
The cells of the gas gland have few mitochondria and moglobin (Root-off shift) and (2)an increase in ionic con-
negligible Krebs cycle activity. For this reason, even in an centration and therefore a reduction in oxygen solubility
oxygen atmosphere, glycolysis in the secretory epithelium (sometimes termed the "salting-out effect"). Both changes
(gas gland) of the swimbladder yields two lactate molecules cause the Po2in the secretory epithelium to increase more
Root-off shift
than that in the swimbladder, so that oxygen diffuses from increased deposition of guanine. Eels turn from yellow to
blood into the gas space of the swim bladder (see Figure silver as a result of these guanine deposits. This occurs as
13-58).The salting-out effect will also reduce the solubility they leave the rivers and begin their migration across
of other gases, such as nitrogen and carbon dioxide, and the oceans.
may explain the high levels of these gases sometimes ob-
served in swim bladders.
Let's return now to the situation in the rete. As dis-
SUMMARY
cussed earlier, erythrocytes are not very permeable to H+ At the level of the mitochondria, the number of oxygen
ions, so the drop in pH in the gas gland is transferred into molecules that an animal extracts from the environment
the red blood cells by CO,, which crosses cell membranes and utilizes is approximately the same as the number of
with ease (Figure 13-59).Acid produced in the gas gland re- carbon dioxide molecules it produces and releases into the
acts with HC0,-, probably taken up from the plasma, pro- environment. In very small animals, gases are transferred
ducing CO, .Thus blood leaving the gas gland and entering between the surface and the mitochondria by diffusion
the venous capillaries of the rete has a high CO, content. As alone, but in larger animals a circulatory system has
the high-CO, venous blood flows through the rete, CO, dif- evolved for the bulk transfer of gases between the respira-
fuses into the arterial blood flowing towards the gas gland. tory surface and the tissues.
This raises the pH of the venous blood, which in turn in- Respiratory surfaces are characterized by large surface
creases oxygen binding by hemoglobin (Root-on shift); as areas and small distances for diffusion between the inhalant
more oxygen is bound, the Po> in the venous blood falls as medium and the blood to facilitate gas transfer. Breathing
it flows away from the gas gland (see Figure 13-58). On the movements assure a continual supply of oxygen and pre-
arterial side of the rete, the entering C 0 2lowers blood pH, vent stagnation of the medium close to the respiratory ep-
which drives oxygen from the hemoglobin (Root-off shift), ithelium. The design of the respiratory surface and the
thereby raising the blood Po2. Thus the Po? changes in the mechanism of breathing are related to the nature of the
rete result from loading and unloading of hemoglobin with medium (i.e., gills in water, lungs in air).
oxygen, with the rete serving as a countercurrent exchanger Bulk transport of 0, and CO, in the blood is aug-
for carbon dioxide and not oxygen. In fact the rete has a mented by the presence of a respiratory pigment (e.g., he-
relatively low oxygen permeability. moglobin). The pigment not only increases the oxygen-
The gas gland and associated rete enable fish to carrying capacity of the blood, but also aids the uptake and
transfer oxygen into the swimbladder even though the release of 0, and CO, at the lungs and tissues.
bladder may contain oxygen at several atmospheres The rate of gas transfer across a respiratory surface de-
pressure. The bladder wall is slightly permeable to gases pends on the ratio of ventilation rate of the respiratory sur-
so there is a continual loss of gas that increases with face to blood flow, ~ I Qas, well as on the absolute venti-
depth (bladder pressure). Gases, therefore, must be se- lation volume and cardiac output. These factors are closely
creted continually to maintain volume in the face of this regulated to maintain adequate rates of gas transfer to meet
loss. Eels migrating at depth across the oceans enlarge the requirements of the tissues. The control system, which
their rete and gas gland and decrease the permeability of has been studied extensively only in mammals, consists of
the bladder wall, enabling them to maintain bladder vol- a number of mechanoreceptors and chemoreceptors that
ume at higher pressures. The permeability of the bladder feed information into a central integrating region, the
wall is decreased by an increase in its thickness due to medullary respiratory center. This center, through a variety
(Table 14-3).There are only minor and transient osmotic salt gland of elasmobranchs or the kidney of mammals.
differences between the intracellular and extracellular flu- The rest of the body surface, with the exception of the lin-
ids of animals. Thus the cell membrane maintains ionic, but ing of the gut, is relatively impermeable to ions and water.
not osmotic, differences between the intracellular and ex- Animals require nutrients and oxygen to maintain me-
tracellular fluids, whereas the epithelium surrounding the tabolism, and as a result of metabolism, they produce waste
body often maintains both ionic and osmotic differences products. Cell membranes that are permeable to oxygen are
between animals and their environments. In most multicel- also permeable to water, and energy must be expended to
lular animals, the entire body surface is not usually in- maintain the ionic and osmotic balance of the animal. An
volved in ionic and osmotic regulation; rather, this regula- animal cannot reduce osmotic and ionic problems by seal-
tion is effected by a specialized portion of the body surface ing itself off from the environment because nutrients must
such as the gills of fish or some internal structure like the be acquired and waste products excreted. Some animals en-
I O N I C A N D OSMOTIC BALANCE 573
...............................................................................
TABLE 14-2
Major inorganic ions of tissues
Ion D~str~butlon Major funct~ons
cyst themselves, but this is feasible only if their metabolic tems, the blood typically passes through excretory organs,
rate is very much reduced. Brine shrimp larvae, for example, generally termed kidneys. In terrestrial animals, the kidneys
can survive in a state of suspended animation for many not only play an important role in the removal of organic
years with little or no growth; when in this state, they can be wastes but are also the primary organs of osmoregulation.
placed in water and revived. This is only possible because
energy turnover is very reduced during encystment, limiting
nutrient utilization and waste-product accumulation. Most
animals, however, do not exist in a state of suspended ani-
mation and must ingest nutrients at a high rate and cope
with the associated osmotic and ionic problems.
The waste products generated during metabolism are
often toxic and cannot accumulate to high levels in the A number of mechanisms are employed to handle os-
body without serious consequences. Thus the cellular envi- motic problems and regulate the differences (1)between in-
ronment must be freed of these toxic by-products of me- tracellular and extracellular compartments and (2)between
tabolism. In the smallest aquatic organisms, this purifica- the extracellular compartment and the external environ-
tion happens simply by diffusion of the wastes into the ment. These are collectively termed osmoregulatory mech-
surrounding water. In animals that have circulatory sys- anisms,a term coined in 1902 by Rudolf Hober to refer to
TABLE 14-3
Electrolyte composition of the human body fluids
Interstitial lntracellular
Serum fluid fluid (muscle)
Electrolytes .
(meq kg-' H,O) .
(meq kg-' H20) (meq kg-' H,O)
Cations
Na+
K+
Ca2+
Mg2+
Totals
Anions
CI- 104 114 2
HC03- 27 31 8
HPO,~- 2 95
sot- 1 20
Organlc aclds 6
Proteln 13 55
Totals 153 145 180
Note: Some of the ions contained within cells are not completely dissolved within the cytosol, but may be partially sequestered within cytoplasmic or-
ganelles. Thus, the true free Ca2+concentration in the cytosol is typically below the overall value given in the table for intracellular Ca2+.Failure of anion
and cation totals to agree reflects incomplete tabulation.
574 INTEGRATION OF PHYSIOLOGICAL SYSTEMS
........................................
the regulation of osmotic pressure and ionic concentrations Regulated osmotic exchanges, which are physiologi-
in the extracellular compartment of the animal body. The cally controlled and serve to aid in maintaining internal
evolution of efficient osmoregulatory mechanisms had ex- homeostasis.
traordinarily far-reaching effects on other aspects of animal
speciation and diversification. The various adaptations and Regulated exchanges generally serve to compensate for
physiological mechanisms evolved by animals to cope with obligatory exchanges. The flux of a substance across a
the rigors of the osmotic environment form especially fas- membrane is determined by its concentration gradient, the
cinating examples of the resourcefulness of evolutionary surface area of the membrane involved, the thickness of the
adaptation. This is the theme of an excellent book by the membrane (i.e. the diffusion distance), and the permeabil-
late Homer Smith entitled From Fish to Philosopher. ity of the membrane. The same factors influence both oblig-
Although there may be hourly and daily variations in atory and regulated exchanges. In the next section, we con-
osmotic balance, an animal is generally in an osmotic sider obligatory exchange and then, in the following
steady state over the long term. That is, on the average, the sections, various mechanisms of regulated exchange.
input and output of water and of salts over an extended pe-
riod are equal. Water enters terrestrial animals with their
food and drink. For animals living in a freshwater envi-
OBLIGATORY EXCHANGE OF IONS
ronment, water enters the body primarily through the res-
AND WATER
piratory epithelium-the gill surfaces of fish and inverte- The integument, respiratory surfaces, and other epithelia in
brates, and the integument of amphibians and many contact with the surrounding milieu act as the barriers to
invertebrates. Water leaves the body in the urine, in the fe- obligatory exchange between an organism and its environ-
ces, and by evaporation through the lungs and integument, ment. The various factors that contribute to the obligatory
the outer covering. exchange are outlined next.
The problem of osmotic regulation does not end with
the intake and output of water. If that were so, osmoregu- Gradients Between the Animal
lation would be a relatively simple matter: A frog sitting in and the Environment
freshwater that is far more dilute than its body fluids would The greater the difference between the concentration of a
merely have to eliminate the same amount of water as substance in the external medium and that in the body flu-
leaked in through its skin, and a camel would just stop ids, the greater the tendency for net diffusion in the direc-
urine production between oases. Osmoregulation also in- tion of the lower concentration. Thus, although a frog im-
volves the maintenance of favorable solute concentrations mersed in a pond tends to take up water from its hypotonic
in the extracellular compartment. Thus, the frog immersed environment, a bony fish in seawater is faced with the prob-
in hypotonic pond water is faced not only with the need to lem of losing water to the surrounding hypertonic seawa-
eliminate excess water, but also with the problem of re- ter. Similarly, a marine fish with a lower NaCl content than
taining salts, which tend to leak out through the skin, be- that of seawater faces a continual diffusion of salt into the
cause the skin in amphibians is generally more permeable body, whereas a freshwater fish faces a continual loss of
than that in the other vertebrate classes. salt. The rate of transfer depends on the size of the gradient
The osmotic exchanges that take place between an an- and the permeability and area of the animal's surface.
imal and its environment can be divided into two classes
(Figure 14-1): Surface-to-VolumeRatio
The volume of an animal varies with the cube of its linear
Obligatory osmotic exchanges, which occur mainly in dimensions, but its surface area varies with the square of its
response to physical factors over which the animal has linear dimensions. Therefore, the surface-to-volume ratio
little or no physiological control is greater for small animals than for large animals. It fol-
. .
I O N I C AND OSMOTIC BALANCE 575
--
of the~rh ~ g hsurface-to-mass (and thus
surface-to-volume) rat~osThls log-log
I
c plot shows the amount of water, as per-
centage of body we~ght,that IS lost per
=.
2
n
o 2-
hour under hot desert cond~t~ons versus
body we~ght [Edney and Nagy, 1976.)
n
-
8
C
.-
C
0
F
0
a
m>
20 1-
(I)
C
F
rn
0
J
70,000g (1.47%)
0
-4 -3 -2 -1 0 1 2 3 4 5
Log body weight (g)
lows that the surface area of the integument, through which have moist, highly permeable skins, through which they ex-
water or a solute can exchange with the environment, is change oxygen and carbon dioxide and through which wa-
greater relative to the water content of a small animal than ter and ions move by passive diffusion. Amphibian skin
it is for a large animal. This means that for a given net rate compensates for loss of electrolytes by active transport of
of exchange across the integument (in moles per second per salts from the aquatic environment into the animal. Fish
square centimeter), a small animal will dehydrate or hy- gills are necessarily permeable, as they engage in the ex-
drate more rapidly than a larger animal of the same shape change of oxygen and carbon dioxide between the blood
(Figure 14-2). and the aqueous environment. The gills, like the frog skin,
also engage in active transport of salts. The volume of
Permeability of the Integument blood perfusing the gills of fish has been shown to decrease
The integument acts as a barrier between the extracellular as respiratory demand drops and to rise in response to in-
compartment and the environment. Movement of water creased oxygen need. This reduction in blood perfusion of
across the integument occurs through cells (transcellular) the gills effectively limits osmotic transfer through the gill
and between cells (paracellular). Pure phospholipid bilay- epithelium during periods of low oxygen uptake. Thus, as
ers, however, are not very permeable to water, and trans- oxygen transfer increases so does osmotic and ionic ex-
cellular movement of water across biological membranes change across the gills.
depends on the presence of water channels. For example, In contrast, reptiles, some desert amphibians, birds, and
erythrocytes swell or shrink rapidly in response to changes many mammals have relatively impermeable skins and thus
in the osmotic strength of the extracellular fluid because of generally lose relatively little water through this route. In
the presence of a 28-kDa protein, appropriately called fact, the skin of some mammals (e.g., cow hide) is so im-
aquaporin. It appears that water channels in membranes permeable that it can be used to carry water or even wine.
are formed of a tetramer of identical aquaporin molecules. The low permeability of the integument of terrestrial ani-
The role of aquaporin as a water channel was demon- mals is maintained in those species that have secondarily
strated in experiments with frogs eggs and oocytes, which become marine or aquatic, such as pond insects and marine
are not very permeable to water and thus do not swell mammals.
much when placed in pond water. When mRNA encoding Not all vertebrates, however, have a relatively imper-
the aquaporin protein was injected into frog oocytes, they meable integument. Many amphibians, as well as mam-
became very water permeable and swelled when placed in mals that perspire, can become dehydrated at low humid-
water. Membrane permeability to water is presumably re- ity because of water loss through the integument. Animals
lated to the concentration of aquaporin water channels with highly permeable skin are simply not able to tolerate
within the phospholipid bilayer. Tight junctions between very hot, dry environments. Most frogs stay close to water.
cells reduce the permeability of the paracellular pathway to Toads and salamanders can venture a bit farther, but they
water. The absence of aquaporin water channels in the also are limited to moist woods or meadows not far from
membrane reduces the transcellular water permeability puddles, streams, or bodies of water in which they can
The permeability of the integument to water and replenish their supply of -body water. These animals also
solutes varies among animal groups. Amphibians generally minimize water loss by behavioral strategies, avoiding
Figure 14-3 The waxy lipid layer on the outside of the insect in-
Epidermis tegument serves as the major water barrier, reducing evaporative
water loss in insects. The waxy layer is deposited through minute
canals in the integument. [Adapted from Edney, 1974.1
desiccation by staying in cool, damp microenvironments contain minute channels that draw water by capillary ac-
during hot, dry times of day. The desert toads, Chiroman- tion to moisten the skin, conserving internal water during
tis xerampelina and Phyllomedusa sauuagii, have extremely cutaneous evaporation.
low evaporative loss of water from their skin because it is Because insects have a waxy cuticle that is highly im-
covered by a secreted wax coating. These toads also excrete permeable to water, their evaporative water loss is much
uric acid rather than ammonia or urea (seethe later section lower than in many other animal groups (Table 14-4).The
Excretion of Nitrogenous Wastes). wax is deposited on the surface of the exoskeleton through
Frogs and toads are endowed with a large-volume lym-
phatic system and an oversized urinary bladder in which
they store water until needed. When these animals wander
from a body of water or during periods of low rainfall, wa-
ter will move osmotically from the lumen of the bladder
into the partially dehydrated interstitial fluid and blood.
The epithelium of the bladder, like the amphibian skin, is
capable of actively transporting sodium and chloride from
the bladder lumen into the body to compensate for the loss
of salts that accompanies excessive hydration during times
of plentiful water. Thus, the anuran bladder serves a dual
function as a water reservoir in times of dehydration and as
a source of salts during times of excessive hydration. The
high water permeability of amphibian skin is used to ad-
vantage to take up water from hyposmotic sources such as
puddles. Many anurans have specialized regions of skin on
the abdomen and thighs, termed the pelvic patch, that 24 28 32 36
when immersed can take up water at a rate of three times Cuticular temperature ("C)
the body weight per day. The permeability of amphibian
skin is controlled by the hormone arginine vasotocin (Am) Figure 14-4 The rate of water loss from insects is much higher at tem-
peratures above the melting point of the waxylayercovering the cuticle.
or more simply uasotocin; like the mammalian hormone
The sharp break in this plot of water loss versus cuticular temperature
vasopressin, or antidiuretic hormone (ADH),vasotocin en- In a cockroach corresponds to the melt~ngpolnt of the waxy cuticle.
hances water permeability. The outer layers of toad skin [From Beament, 1958.1
I O N I C A N D OSMOTIC BALANCE 577
......................................
TABLE 14-4 though water is another end product of cellular metabolism,
Evaporative water loss of representative animals under desert it is produced in small enough quantities that its elimination
conditions
is no problem (Table 14-5).In fact, this so-called metabolic
Water loss water is the major source of water for many desert dwellers.
Species (mg .cm-' h-') Remarks* Osmotic problems are posed by the inevitable production
- -- -
70
Figure 14-5 Water loss associated with respiration de-
60 pends on the relationship between the body tempera-
Lung air saturated ture and the inhaled-air temperature, as well as on the
- 4050
.k
m
relative humidity of the inspired air. As unsaturated air is
warmed in the lungs, ~ t spicks up moisture until it is satu-
rated (gray bars). During exhalation, the air is cooled in
F 20
when the inhaled air is at 25% relative humidity (r.h.1and
15°C (left bars) or 30°C (right bars). Clearly, the amount
10 of water recovered is greaterand hence the amount lost
is less when the inhaled air is at the lower temperature.
0 Indeed, under these climatic conditions, the kangaroo
10 20 30 40 rat exhales air at 13'C (lower than ambient!). [Adapted
Ambient air temperature from Schmidt-Nielsen et al., 1970.1
I O N I C A N D OSMOTIC BALANCE 579
...............................................................................
The respiratory loss of water is minimized through a lium (Figure 14-6B). Mammals, Including humans, em-
mechanism first discovered in the nose of the desert- ploying this mechanism to humidify inhalant air have
dwelling kangaroo rat, Dipodomys merriami, by Knut "cold" noses, which can be wet or even occasionally drip.
Schmidt-Nielsen. This mechanism, termed a temporal With the next inhalation, this condensed moisture again
countercurrent system, retains most of the respiratory contributes to the humidification of the inspired air, and the
water vapor by condensing it on cooled nasal passages dur- cycle is repeated, most of the vapor being recycled within
ing expiration. Air entering the nasal passages is warmed to the respiratory tract.
about 37-38°C and humidified by heat and moisture ab- The nose, therefore, plays an important role in reduc-
sorbed from the tissues of the nasal passages, trachea, and ing the loss of water and heat from the body. The impor-
bronchi (Figure 14-6A). The nasal passages are cooled by tance of the nose in cooling expired air can be detected eas-
this evaporative water loss and the flow of cool air through ily by placing your hand in front of your nose and mouth
the nasal passages. The tissue temperature is lowest at the and breathing out via your mouth and via your nose; the
tip of the nose and increases along the nasal passages to- temperature difference is usually obvious. Because there is
wards the lung. The nose has a large blood supply to main- little cooling of air expired through the mouth, ,the loss of
tain the delivery of water to humidify the incoming air. The water and heat is greater when breathing out via the mouth
blood supply does not warm the nose because it is arranged (e.g., when the nose is clogged due to a cold) compared
in a countercurrent fashion, so that warm blood entering with expiration through the nose. If air flow through the
the nasal region is cooled by cold blood leaving the nose. nasal passages is bypassed by placing a tube in the trachea,
During exhalation, the process of heat exchange be- as during human or animal surgical operations, heat and.
tween air and nasal tissues is reversed. The warm expired water loss may increase; and surgical patients must be given
air is cooled to somewhat above ambient as it passes back additional food and water to compensate for the increased
out through the nasal passages, which had been cooled by water loss. The increased water loss from the trachea can
the same air during inhalation. As the expired air gives up contribute to post-surgical sore throat, a common problem.
some of its heat to the tissues of the nasal passages, most of A similar mechanism for trapping exhaled moisture oc-
the acquired moisture condenses on the cool nasal epithe- curs in numerous birds and lizards. Where salt glands drain
A Inspiration B Expiration
I
Outgo~ngair is cooled
and loses water, wetting
Air
Figure 14-6 Temporal countercurrent exchange in the respiratory sys- sages. (B) During expiration, the same air loses most of the heat and wa-
terns of many vertebrates acts to conserve body heat and body water. ter it gained earlier, as it warms and deposits water on the cooled nasal
(A) During inspiration, cool air (e.g., at 28°C) is warmed and humidified passages on its way out. Small red arrows indicate direction of heat and
as it flows toward the lungs, removing heat and water from the nasal pas- water movement; long arrows indicate direction of air flow.
580 INTEGRATION OF PHYSIOLOGICAL SYSTEMS
.......................................
into the nasal passages, as they do in iguanas, the water in from marine invertebrates continue to function for many
the excreted salt solution enters the incoming air during in- hours when placed in seawater. Ionic concentrations in the
spiration and is largely conserved by being condensed dur- body fluids of freshwater and terrestrial invertebrates differ
ing exhalation. Occassionally all the water from the salt from seawater; in these animals the body fluids are invari-
solution is evaporated, leaving salt deposits around the ably more dilute than seawater but considerably more con-
nose of these seawater-drinking lizards. This will occur, centrated than freshwater.
however, only if the animal's temperature is above that of Some aquatic invertebrates are strict osmoregulators
the environment. like the vertebrates, some are limited osmoregulators, and
Water loss via the lungs is small for mammals living in some are strict osmoconformers. These classes are illus-
hot humid climates and large for those living in cold dry cli- trated in Figure 14-7, in which the osmolarity of the
mates. The rate of ventilation and the ventilation pattern extracellular compartment is plotted against the osmolar-
(e.g., breathing through either the mouth or nose) also af- ity of the aqueous environment. As the osmolarity of the
fect the rate of water loss via the lungs. The problem of res- environment changes, the osmolarity of a strict osmo-
piratory water loss is much less in animals with body tem- conformer changes by an equal amount, paralleling the line
peratures similar to the ambient temperature; in this case that describes internal-external osmolar equality. In con-
the air has only to be humidified at ambient temperature. A trast, a strict osmoregulator maintains a constant internal
reptile with a low metabolic rate and, therefore, a low vefi- osmolarity over a large range of external osmolarities, so as
tilation rate and with a body temperature equal to the am- to produce a horizontal plot parallel with the abscissa. Lim-
bient temperature will have only minimal rates of water ited osmoregulators regulate over a limited range of osmo-
loss via the lung. This gives reptiles a n advantage over larities and conform at other environmental osmolarities.
mammals in regions where water is scarce.
Marine
teleost
Hypoton~c Isotonic
- --_
Secretes salt
seawater
Drinks
from gills
Freshwater Strongly
Hypertonic Drinks no water
teleost hypotonic
Amph~blan Hyperton~c
Strongly
hypotonic @ Absorbs salts through s k ~ r
Marine
rept~le
Hypotonic Isotonic
- Drinks seawater
Desert
mammal
Marine
Hypotonic
Strongly
hypertonic
Strongly
a secretion
Drinks no water
Depends on
metabolic water
Does not
mammal hypertonic drink seawater
c Drinks seawater
Marine Weakly
bird hypertonic 0
Hypertonic salt-gland
Weakly secretion
hypertonic
urine
b Terrestrial Weakly
bird hypertonic
Figure 14-8 Animals living in different environments exhibit various os- some vertebrates is illustrated here. Passive loss of water through the
moregulatory mechanisms. The active exchange of water and salt in skin, lungs, and alimentary tract is not indicated.
significantly lower concentrations than they are in seawa- may have evolved to accommodate the requirements of
ter, whereas Naf and C1- are higher. Since various func- neuromuscular function.
tions of excitable tissues such as nerve and muscle of ver- Like the hagfish, the cartilaginous fishes such as sharks,
tebrates are especially sensitive to the concentrations of rays, and skates, as well as the primitive coelacanth La-
Ca2+ and Mg2+,the regulation of these divalent cations b timeria, have plasma that is approximately isosmotic to
I O N I C A N D OSMOTIC BALANCE 583
...............................................................................
Food +
only
Figure 14-9 Salt and water exchange is in the opposite direction in fresh- salts. They achieve this by drinking seawater and then eliminating salts by
water and marine teleosts. (A) Freshwater teleosts prevent the net gain several routes. Solid arrows indicate active processes; broken arrows, pas-
of water and loss of salt by copious excretion of a dilute urine from which sive processes. Note the active role of the gills In salt transport in both
most of the salt has been reabsorbed. (B) Marine teleosts face the op- groups. [Adapted from Prosser, 1973.1
posite osmotic problems, namely, avoiding a water deficit and excess of
seawater. They differ, however, in that they maintain far rine teleosts, the gill epithelium contains specialized cells,
lower concentrations of electrolytes (i.e., inorganic ions), called chloride cells, that mediate transport of NaCl from
making up the difference with organic osmolytes such as the blood into the surrounding water. The mechanism of
urea and trimethylamine oxide (TMAO).High urea con- this transport, which makes it possible for these fishes to
centrations tend to cause the breakup of proteins into con- live in saltwater; is described in a later section.
stituent subunits, whereas TMAO has the opposite effect,
canceling the effect, of urea and stabilizing protein structure
in the face of high urea levels. In the elasmobranchs and
coelacanths, excess inorganic electrolytes such as NaCl are
excreted via the kidneys and also by means of a special ex-
cretory organ, the rectal gland, located at the end of the al-
imentary canal.
The body fluids of marine teleosts (modern bony
fishes), like those of most higher vertebrates, are hypotonic
to seawater, so there is a tendency for these fishes to lose Some teleost species-for example, the salmon of the
water to the environment, especially across the gill epithe- Pacific Northwest and eels in eastern North America and
lium. To replace the lost volume of water, they drink salt- Europe-are able to maintain a more-or-less constant
water. Most of the net salt uptake is due to drinking sea- plasma osmolarity even though they migrate between ma-
water rather than salt uptake across the body surface or rine and freshwater environments. Such fish undergo a
gills. By absorption across the intestinal epithelium, 70% physiological adaptatlon that enable them to maintain a
to 80% of the ingested water enters the bloodstream, along more-or-less constant ionic composition In both environ-
with most of the NaCl and KC1 in the seawater. Initially the ments. Some of the physiological changes that occur when
ingested seawater is diluted by about 50% by diffusional teleosts migrate from freshwater to seawater begin before
uptake of salts across the esophagus. Active salt uptake oc- the animal enters seawater. Eels, for instance, reduce the
curs in the small intestine, via a Na/2Cl/K cotransporter permeability of the integument, changing from yellow
across the apical membrane and then by active transport to silver in the process. Likewise, the gill reorganization
via a Na+/K+ ATPase across the basolateral membrane. that characterizes adaptation of salmon to seawater begins
Left behind in the gut and expelled through the anus are as the fish migrate down river to the ocean. We'll discuss
most of the divalent cations such as Ca2+, Mg2-, and the adaptation of migrating teleosts in detail later in this
SO:-. The excess salt absorbed along with the water is chapter.
subsequently eliniinated from the blood by active transport To summarize, freshwater animals tend to take in wa-
of Na+, C1-, and some K+ across the gill epithelium into the ter passively and to remove it actively through the osmotic
seawater, and by secretion of divalent salts by the kidney work of kidneys (vertebrates)or kidney-like organs (inver-
(see Figure 14-9B).The urine is isotonic to the blood, but tebrates). They lose salts to the dilute environment and re-
rich in those salts (especiallyCa2+,Mg2+,and SO:-) that place them by actively absorbing ions from the surround-
are not secreted by the gills. The net result of the combined ing fluids into their bodies through skin, gills, or other
osmotic work of gills and kidneys in the marine teleost is a actively transporting epithelia. Marine fish, on the other
net retention of water. hand, lose water osmotically through the gills or through
The gills of marine teleosts, as might be exoected, are the integument, if it is permeable. To replace lost water, ma-
organized differently from those of freshwater fish. ma- rine fish drink seawater and actively secrete the excess
salt ingested with the seawater back into the environment.
This process takes place through active transport in ex-
trarenal osmoregulatory organs such as gills and the rectal
gland.
Air-Breathing Animals
Animals in a terrestrial environment can be thought of
as submerged in an ocean of air rather than water. Unless
the humidity of the air is high, animals having a water-
permeable epithelium will be subject to dehydration very
much as if they were submerged in a hypertonic medium
such as seawater. Dehydration would be avoided if all ep-
ithelial surfaces exposed to air were totally impermeable
to water. The evolutionary process has not found this to
be a feasible solution to the problem of desiccation, since
an epithelium that is impermeable to water (and thus dry)
will have limited permeability to oxygen and carbon diox-
ide, and will thus be unsuited for the respiratory needs of Seawater Nasal fluid Urine
(3%salt) (5% salt) (3%salt)
a terrestrial animal. As a consequence, air-breathing ani-
mals are subject to dehydration through their respiratory
epithelia. Air-breathing animals utilize various means to
minimize water loss into the air by this route and others
(see Figure 14-8).
Marine reptiles (e.g., iguanas, estuarine sea turtles,
crocodiles, sea snakes) and marine birds drink seawater to
obtain a supply of water but, like marine teleosts, are un-
able to produce a concentrated urine that is significantlyhy-
perosmotic to their body fluids. Instead, they are endowed
with glands specialized for the secretion of salts in a strong
hyperosmotic fluid. These salt glands are generally located
above the orbit of the eye in birds and near the nose or eyes
in lizards. Brackish-water crocodiles were long suspected of Seawater Ur~ne
(3%salt) (2'%salt)
using extrarenal means of excreting salts, and eventually
salt glands were discovered in the tongue of this reptile. Al- Figure 14-10 Marine reptiles and birds drink seawater to obtain water,
though neither reptilian nor avian kidneys are capable of whereas most mammals become dehydrated if they drink seawater. (A)
producing a very hypertonic urine, the salt glands of marine When marine birds drinkseawater,they secrete NaCl via the salt glands,
thereby eliminating 80% of the ingested salt along with only 50% of the
reptiles and birds secrete a sufficiently concentrated salt so- ingested water. As a result, these birds can produce a hypotonic urine
lution to enable them to drink saltwater even though their without dehydrating. (B) When humans and other mammals, who lack
kidneys are unable to produce urine more concentrated salt glands, drink seawater, they cannot concentrate the urine sufficiently
than seawater (Figure 14-10A). Salt glands compensate in t o conserve water while eliminating the ingested salt. Like terrestrial
these groups for the inability of their kidney to produce a mammals, marine mammals cannot drink seawater; these animals use
various water-conservation mechanisms to survive. [From "Salt Glands"
urine that is strongly hypertonic relative to body fluids. Ma-
by K. Schmldt-Nielsen,Copyright 0 1959 by Scientific American, Inc. All
rine mammals, which lack salt glands or similar specializa- r~ghtsreserved.] I
tions, avoid drinking seawater, get their water entirely from
their food intake and its subsequent metabolism, and de-
pend primarily on their kidneys for maintaining osmotic
balance. tion. Thus humans stranded at sea will die unless they have
Human beings, like other mammals, are not equipped access to freshwater. They cannot replace lost water by
to drink seawater. The human kidney can remove up to drinking seawater. If they do so, it will only make the prob-
about 6 g of Nat from the bloodstream per liter of urine lem worse. Humans require a constant source of fresh
produced. Because seawater contains about 12 g.L-' of drinking water to excrete accumulated salts and metabolic
Na+, imbibing seawater will cause a human being to accu- waste products.
mulate salt without adding a physiologically equivalent Mammals cannot drink seawater, and yet marine mam-
amount of water (Figure 14-10B).Stated differently, to ex- mals such as pinnipeds (e.g., sea lions, seals) and cetaceans
crete the salt ingested with a given volume of seawater, the (e.g., porpoises, whales) live in the ocean, even though they
human kidney must pass more water than is contained in do not have extrarenal salt-secreting organs like the salt
that volume. This, of course, will lead rapidly to dehydra- glands of birds and reptiles. Camels and many other mam-
IONIC AND OSMOTIC BALANCE 585
...............................................................................
mals can survive in the desert. Thus, unlike humans, many abruptly, since the cooling properties of the nasal epithe-
mammals can survive in habitats where drinking water is lium will be reduced. Desert mammals also generally avoid
not available. Joseph Priestley (1733- 1804),who was the heat-generating exercise during the day, when removal of
first to isolate many gases, including oxygen, observed that excess heat from the body is slowed by the higher ambient
he could keep mice alive without water for three or four temperature. Because of its efficient kidneys, the kangaroo
months, in a cage on a shelf above the kitchen fireplace in rat excretes a highly concentrated urine, and rectal absorp-
his home in Yorkshire, England. That is, he observed that tion of water from the feces results in essentially dry fecal
mice could survive for many months without drinking wa- pellets.
ter. Another small mammal, the kangaroo rat, Dipodomys By using all these adaptations for desert survival, the
merriami, a native of the American Southwest, has become kangaroo rat greatly reduces its potential water loss. In
a classic example of how small mammals survive without spite of this extreme osmotic economy, the small amount of
drinking water in the arid conditions of the desert. Let's see lost water must, of course, be replaced, or the animal will
how these mammals, as well as certain terrestrial arthro- eventually dry up. Since the kangaroo rat eats dry seeds that
pods, survive in the absence of freshwater. contain only a trace of free water, is not known to drink,
and in fact survives quite well in the near absence of free
Desert-living mammals water, it must have a cryptic source of water. This, it turns
The survival strategies practiced by the kangaroo-rat ex- out, is the metabolic water noted earlier (see Table 14-5).
emplify a variety of osmoregulatory adaptations charac- The exquisite conservation of water by the kangaroo rat al-
teristic of many small desert mammals (Figure 14-11).The lows it to survive primarily on the water produced during
kangaroo rat and other desert mammals are faced with a the oxidation of foods, so that over the long term water
physiological double jeopardy-excessive heat and near gains equal water losses (Table 14-7).
absence of free freshwater. Water regulation and tempera- Unlike the kangaroo rat, camels are too large to hide
ture regulation are, of course, closely related, since one im- from the hot desert sun in burrows. When deprived of
portant means of channeling excess heat out of the body drinking water, camels do not sweat but allow their body
into the environment is by evaporative cooling. Since evap- temperature to rise rather than losing water by evaporative
orative cooling is at odds with water conservation, most cooling during the heat of the day. During the cooler night,
desert animals cannot afford this method and have devised the camel's body temperature drops and increases only
means of circumventing it. The kangaroo rat, like many slowly the next day because of the animal's large body mass
desert mammals, avoids much of the daytime heat by re-
maining in a burrow during daylight hours and coming out
TABLE 14-7
only at night. This nocturnal lifestyle is an important and Sources of water gain and loss by the kangaroo rat
widespread behavioral adaptation to desert life. Not only
does the cool burrow reduce the animal's temperature load, Gains Losses
but it reduces respiratory water loss. The nasal counter- Metabol~c 90% Evaporat~on+ 70%
current mechanism for conserving respiratory moisture de- water persp~rat~on
pends, of course, on the ambient temperature in the burrow Free water In
"dry" food 10% Ur~ne 25%
being significantly lower than the 37°C to 40°C character-
Dr~nk~ng 0%
- Feces 5%
-
istic of the core temperatures of birds and mammals. If
the rodent ventures out of its cool burrow into air close to 100% 100%
its own temperature, its respiratory water loss will rise Source Schm~dt-N~elsen,
1972
ndensed in nasa
es dehydrated prior
586 INTEGRATION OF PHYSIOLOGICAL SYSTEMS
...............................................................................
Dehydrated
42
-e
-$s
a,
40
n
Watered -
>.
-0
38
37 CS; 36
36 r V
I
camel
I I
34 1
6 A.M. 12 6 PM.
Time (days) Time
Figure 14-12 When water is scarce, large desert animals such as the the use of water for temperature regulation. (B) Diagrammatic represen-
camel exhibit a large, but slow, increase in body temperature during the tation of the daily patterns of body temperature in a large and a small
day, whereas smaller animals heat up rapidly when exposed to the sun. mammal subjected to heat stress under desert conditions. Small animals
(A)The daily temperature fluctuation in a well-watered camel and in a de- must enter burrows per~odicallyto avoid overheating. [Part A from
hydrated one. When the camel is depr~vedof drinking water, the daily Schmidt-Nielsen, 1963; part B from Bartholomew, 1964.1
fluctuation may increase to as much as 7°C. This has a great influence on
and thick fur, which acts as a heat shield. Nevertheless, flow through the blow hole is high because both inspiration
the body temperature of a dehydrated camel may vary from and expiration are rapid, and large volumes of gas are
35°C at night to 41°C during the day (Figure 14-12A). moved with each breath. It is possible that the expansion of
This strategy of heating during the day and cooling at air passing through the blow hole of a whale also cools the
night is impossible in small rodents, whose body tempera- air, resulting in water condensation in the region of the
tures oscillate much more rapidly than in the larger camel blow hole that can be used to wet inspired air. This would
(Figure 14-12B).Because of their small size, desert rodents reduce water loss via ventilation.
heat up- rapidly
- . in the sun and must return to their burrow
to cool down. The camel also reduces heating by orienting
to give minimal surface exposure to direct sunlight. The
camel, like other desert animals, produces dry feces and
concentrated urine. When water is not available the camel
does not produce urine but stores urea in the tissues. The
camel can tolerate not only dehydration but also high - urea
levels in the body. When water becomes available, these
ships of the desert can rehydrate by drinking 80 liters in A remarkable example of water retention in a marine
10 minutes. mammal faced with desert-like problems of water conser-
vation occurs in a recently weaned baby elephant seal.
Marine mammals After being abandoned by its mother, the baby seal goes for
Marine mammals face problems similar to those of desert 8-10 weeks without food or water. During this time the
animals because they live in an environment without avail- baby seal's only source of water is that derived from the ox-
&to
--- - -
able drinking water. E e r . water everywhere and not a
drink! The physiological responses of marine mam-
mals, although different in detail, are generally similar to
idation of its body fat. It weighs about 140 kg at the time
of weaning and loses only about 800 g of water per day, of
which less than 500 g is lost through respiration. This econ-
those of desert mammals. The emphasis is on water con- omy is ascribed both to its nasal countercurrent heat ex-
servation. They are endowed, as are other mammals, with changer and to a slowing of metabolic rate, which allows
highly efficient kidneys capable of producing a very hyper- it to stop breathing for 40 minutes and then alternate with
tonic urine. Seals have a characteristic labyrinth-like pro- 5 minutes of deep breathing. The ability to suspend breath-
liferation of epithelial surfaces in the nasal passages which ing is, of course, not uncommon for marine mammals such
reduces water loss via breathing. Whales and dolphins have as the elephant seal, which can dive for prolonged periods.
a blow hole rather than the typical mammalian nose. These The ability to conserve water is also seen in the adult ele-
animals have large lung tidal volumes. The velocity of air phant seal. The large males spend up to three months on
I O N I C A N D OSMOTIC BALANCE 587
...................................
the beach and when on land do not drink or eat. The fe- epithelial cells composing these epithelia differ from all
males suckle their young for about four weeks on the beach other cell types in being anatomically and functionally po-
and then the pup is abandoned when the female goes to sea larized. The apical surface (sometimes referred to as the
for four months. She returns for about a month to molt and mucosal or luminal surface) of an epithelial cell faces a
then goes to sea for another six months. While at sea, the space that is continuous with the external world (the sea,
female elephant seal does not drink but relies on the water the pond, the lumen of the gut, the lumen of a kidney
in fish in her diet and metabolic water to maintain her wa- tubule, etc.). The other side of the epithelial cell, the basal
ter requirements. surface (sometimes referred to as the serosal surface) gen-
erally bears deep basal clefts and faces the internal com-
Terrestrial arthropods partment containing extracellular fluid. This internal com-
Certain terrestrial arthropods have the ability to extract partment is the one that contains all the other cells of the
water vapor directly from the air, even, in some species, remaining body tissues. These exist, so to speak, in their
when the relative humidity is as low as 50% (Table 14-8). own private "pond," composed of the extracellular fluid in
To date, this poorly understood ability has been demon- which they are bathed. The proper composition of this in-
strated only in certain arachnids (ticks, mites) and in a ternal pond depends on the osmoregulatory work and bar-
number of wingless forms of insects, primarily larvae. rier functions performed by epithelial cells.
Those species that exhibit this ability live in habitats devoid The excretion of nitrogenous wastes varies among
or nearly devoid of free water. The ability to remove water species depending on water availability. The nature of the
from the air is all the more remarkable in these arthropods nitrogenous end product varies and many different organs
because it normally occurs even when the vapor pressure of are involved in the excretion of ammonia, urea, andlor uric
the hemolymph exceeds that of the air, which it does at all acid. In freshwater fish, for example, ammonia is usually
values of relative humidity below 99%. the major nitrogenous end product and the gills are the ma-
jor site of excretion. In mammals, by contrast, the major ni-
TABLE 14-8
Critical equilibrium humidities for some arthropods that can trogenous end product is urea and the kidneys the site of
extract water from the vapor phase excretion. Because the excretion of nitrogenous end prod-
ucts is variable and not organ specific, it is discussed at the
Relatlve
hum~dlty end of this chapter.
The mechanisms for transporting substances across ep-
Arachnida ithelia were discussed in Chapter 4 and the same basic cel-
lxodes ricinus 92 0
Rhiprcephalus sangurneus 84 0-90 0 lular machinery is used in all excretory or osmoregulatory
lnsecta organs. For example, similar salt-excreting cells are found
Thermobia domestrca 45 0 in the nasal gland of birds and reptiles, the mammalian kid-
Tenebrio molrtar larvae 88 0 ney, the rectal gland of elasmobranchs, and the gills of ma-
Note At relatlve hum~dltlesbelow crltlcal, the an~malIS unable to extract rine teleost fishes. Not only are the cells similar but their ac-
molsture from the alr tivities are regulated by similar arrays of hormones. The
Source Edney and Nagy, 1976 detailed functioning of organs with similar cellular struc-
ture may be different because of the gross organization of
The water vapor pressure associated with a solution de- the organ. The capabilities of transport epithelia are greatly
creases with increasing ionic content, so highly concen- enhanced in osmoregulatory organs by their anatomic or-
trated salt solutions will absorb water from air. Insects take ganization, as is exquisitely evident in the kidneys of mam-
advantage of this by creating very concentrated solutions mals. Here, in addition to a high degree of cellular differ-
that can absorb water from air. In insects that extract wa- entiation for transepithelial transport, the epithelium is
ter from air, the site of entry is often the rectum, which re- organized into tubules arranged so as to enhance the trans-
duces the water content of fecal matter to remarkably low port efficiency of the tubular epithelium. This combination
levels. As water is removed from the feces, the latter can of cell function and tissue organization has produced a
take on new water from the air, if the water vapor pressure marvelously efficient osmoregulatory and excretory organ.
is high enough and if the air has access to the rectal lumen. The next several sections describe and compare the opera-
In ticks, tissues in the mouth have been implicated in the tion of various types of osmoregulatory organs found in
uptake of water vapor. Here it appears that the salivary different animals.
glands secrete a highly concentrated solution of KC1 that in
turn absorbs water from the air.
MAMMALIAN KIDNEY
The mammalian kidney is the osmoregulatory organ of
OSMOREGULATORY ORGANS which we have the most complete understanding, thanks to
The osmoregulatory capabilities of metazoans depend to intensive research over the past four to five decades. The
a great extent on the properties of transport epithelia lo- mammalian kidney performs certain functions that in
cated in gills, skin, kidneys, and gut. The highly specialized lower vertebrates are shared by other organs such as the
......................................
skin and bladder of amphibians, the gills of fishes, and the Renal
salt glands of reptiles and birds. Thus, the mammalian kid-
ney is not representative of all vertebrate kidneys, which
are organized somewhat differently in different groups of
vertebrates.
paracellular diffusion between the lumen and interstitial border apical side or
luminal side
space surrounding the renal tubule. \
The nephron can be divided into three main regions:
the proximal nephron, the loop of Henle, and the distal
nephron. The proximal nephron consists of Bowman's cap-
sule and the proximal tubule. The hairpin loop of Henle
comprises a descending limb and an ascending limb. The
latter merges into the distal tubule, which joins a collecting
duct serving several nephrons. The nun-ber of nephrons per
kidney varies from several hundred in lower vertebrates to
many thousands in small mammals, and a million or more
in humans and other large species.
The loop of Henle, found only in the kidneys of birds
and mammals, is considered to be of central importance in
concentrating the urine. Vertebrates that lack the loop of
Henle are incapable of producing a urine that is hyperos-
motic to the blood. In mammals, the nephron is so oriented
that the loop of Henle and the collecting duct lie parallel to Serosal side or
each other (see Figure 14-14).The glomeruli are found in basolateral side or
the renal cortex, and the loops of Henle reach down into blood side
the papillae of the medulla; thus the nephrons are arranged Figure 14-15 The cells of the proximal tubule are specialized for the
in a radiating fashion within the kidney (seeFigure 14-13). transport of salts and other substances from the luminal (apical) side to
Nephrons can be divided into two groups: the serosal (blood) side. The apical membranefacing the lumen is thrown
into fingerlike projections (microvilli),greatly increasing its surface area.
This surface is referred to as a brush border. Mitochondria are concen-
Juxtamedullary nephrons, which have their glomeruli
trated near the basolateral (serosal) surface, which is thrown into deep
in the inner part of the cortex and long loops of Henle basal clefts. These features allow the concentration of salts in the renal in-
that plunge deeply into the medulla (seeFigure 14-14A) terstitium by active transport of salts across the basal membrane.
590 INTEGRATION OF PHYSIOLOGICAL SYSTEMS .
...............................................................................
Figure 14-16 Blood pressure in the renal
glomerulus is high because of the low-resis-
tance input pathway (afferentarteriole) and the
high-resistanceoutput pathway. Regulation of
glomerular blood pressure, which influences
the filtration rate, is largely through modulation
of the diameter of the afferent arteriole.
Bowman's capsule
Cortical nephrons, which have their glomeruli in the Tubular reabsorption of approximately 99% of the wa-
outer cortex and relatively short loops of Henle that ex- ter and most of the salts from the ultrafiltrate leaving
tend only a short distance into the medulla (see Figure behind and concentrating waste products such as urea
14-14B) Tubular secretion of a number of substances via active
transport in nearly all instances
The anatomy of the renal circulatory system is impor-
tant in the function of the nephron. The renal artery subdi-
Formation of the ultrafiltrate is the initial step in urine pro-
vides to form a series of short afferent arterioles, one of
duction; reabsorption and secretion occur along the length
which supplies each nephron (see Figure 14-14). The
of the renal tubule. In addition to these processes, the ex-
glomerular capillaries within Bowman's capsule are sub-
cretion of nitrogenous wastes, discussed at the end of the
jected to somewhat higher pressures than other capillaries
because of the low-resistance input pathway and high-
resistance output pathway (Figure 14-16).The capillaries
of the glomerulus recombine to form an efferent arteriole.
Unlike most other vessels, which would join to form veins,
the efferent arteriole in juxtamedullary nephrons then sub-
divides again to form a second series of capillaries sur-
rounding the loop of Henle. Thus the blood, on leaving the
glomerulus located in the cortex, enters the efferent arteri-
ole and is carried into the medulla in a descending and sub-
sequently ascending loop of anastomosing (interconnect-
ing) capillaries before leaving the kidney via a vein. The
hairpin loops, which parallel the loops of Henle of jux-
tamedullary nephrons, are referred to as the vasa recta (see
Figure 14-14A).Flow in the efferent arteriole is less than
that in the afferent arteriole because around 10% of
the blood is filtered across the Bowman's capsule. For hu-
mans this amounts to about a liter of filtrate formed every
10 minutes. Clearly urine flow rate is much less, so much of
the initial filtrate formed in the Bowman's capsule is reab-
sorbed into the blood across the kidney tubule.
Urine Production
Urine
Three main processes contribute to the ultimate composi-
Figure 14-17 Urineformation in the mammalian nephron involvesthree
tion of the urine (Figure 14-17):
main processes. Filtration, the initial step, takes place in Bowman's cap-
sule, followed by reabsorption and secretion, which occur along the re-
Glomerular filtration of plasma to form an ultrafiltrate nal tubule. The final product of these processes is a hypertonic urine,
in the lumen of the Bowman's capsule whose composition differs from that of blood.
chapter, involves the synthesis of certain excreted products and the lumen of Bowman's capsule, which favors filtra-
in the tubular cells and lumen. tion; (2)the colloid osmotic pressure, which opposes filtra-
tion; and (3) the hydraulic permeability (sievelikeproper-
Glomerular filtration ties) of the three-layered tissue separating these two
The glomerular ultrafiltrate contains essentially all the con- compartments. The net pressure gradient results from the
stituents of the blood except for blood cells and nearly all sum of the hydrostatic pressure difference between the two
blood proteins. Filtration in the glomerulus is so extensive compartments and the colloid osmotic pressure difference.
that 15%-25% of the water and solutes are removed from The latter arises because of the separation of proteins dur-
the plasma that flows through it. The glomerular ultrafiltrate ing the filtration process. In humans, the proteins remain-
is produced atthe rate of about 125 rnl-min-l, or about 180 ing in the capillary plasma produce an osmotic pressure
.
L day-', in human kidneys. When this number is compared difference of about -30 mm Hg, and the hydrostatic pres-
to the normal intake of water, it is evident that unless most sure difference (capillary blood pressure minus the back
of the glomerular ultrafiltrate is subsequently reabsorbed pressure in the lumen of the Bowman's capsule) is about
into the bloodstream, the body would be quickly dehy- +40 mm Hg (Table 14-9).The result is a net filtration pres-
drated, thus much of the ultrafiltrate must be reabsorbed. sure of only about + 10 mm Hg. This small pressure differ-
ential acting on the high permeability of the glomerular
sieve produces a phenomenal rate of ultrafiltrate formation
by the millions of glomeruli in each human kidney. It is im-
portant to note that the filtration process in the kidney is
entirely passive, depending on hydrostatic pressure that de-
rives its energy from the contractions of the heart. In lower
vertebrates such as the salamander, the blood pressure in
the glomerular capillaries is much lower than in humans,
but the net filtration pressure is not that much less than in
the human kidney, because of a lower intracapsular and os-
The process of ultrafiltration in the glomerulus motic pressure in the salamander (see Table 14-9).
(Figure 14-18)depends on three factors: (1)the net hydro-
static pressure ddference between the lumen of the capillary TABLE 14-9
Balance sheet o f pressures (in m m Hg) involved in glomerular
ultrafiltration as illustrated i n Figure 14-20
Afferent Efferent arteriole Salamandar Man
arteriole I
Capillary Glomerular cap~llarypressure 17 7 55
pressure, lntracapsular pressure -1 5 -15
+55 rnrn Hg - -
Net hydrostatic pressure 162 40
Colloid osmotlc pressure -104
- - 30
-
Net f~ltrat~on
pressure 58 10
Source Pltts, 1968, Brenneret al , 1971
Collo~dosmot~c
,Filtration
slits
7 Endothelial
pores
-Endothelium
of
glomerulus
Basement membrane
of glomerular capillary
Figure 14-19 The inner (visceral)surface of Bowman's capsule is spe- largement of the portion of partA that is enclosed in a box. Substances
cialized for filtering the blood in the glomerular capillaries (A) Overview pass from the blood through the endothelial pores, across the basement
of glomerulus. The podocytes composing the visceral layer have long membrane, and then through the filtration slits between pedicels.
processes, termed pedicels, which cover the vascular epithelium. (B) En-
spaces, the filtration slits, between them (Figure 14-19B). based mainly on molecular size, but also on shape and
The filtrate, driven by the net pressure drop across the en- charge (Table 14-10).There is a bulk flow of water through
dothelium, passes through the pores formed in the walls of the sieve carrying with it ions, glucose, urea, and many
the glomerular capillaries and then through the filtration other small molecules.
slits. The three-layered membrane separating the capillary The kidneys are perfused by 500-600 ml of plasma per
lumen and lumen of Bowman's capsule acts as a molecu- minute, or 20%-25% of the cardiac output, yet constitute
lar sieve, excluding almost all proteins from the ultrafiltrate less than 1% of the body weight. This preferential perfu-
TABLE 14-10
Relation between the molecular size of a substance and the ratio of its concentration in the filtrate appearing in Bowman's capsule t o
its concentration in the plasma, [filtrate]/[filtrand]
D~mens~ons
(In nanometers)
Water 18 0.1 1 10
Urea 6 0.16 1.O
Glucose 180 0.36 1.O
Sucrose
Insulin
Myoglobin
Egg albumin
Hemoglobin
Serum albumin
Source P~tts,1968
I O N I C A N D OSMOTIC BALANCE 593
. . . . . . . . . . . . .< . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
sion takes place in a relatively low-resistance vascular bed pends on a number of interrelated mechanisms involving
within the kidney. A high renal blood pressure is the result both paracrine and endocrine secretions as well as neural
of the relatively direct arterial supply; because arteries and control.
arterioles are large in diameter and short in length, the loss Several intrinsic mechanisms provide autoregulation of
of pressure due to friction is minimized. The efferent arte- the glomerular filtration rate. First, an increase in blood
rioles (those taking the blood away from the glomeruli) are pressure will tend to stretch the afferent arteriole, which
of smaller diameter and along with the capillaries of the would be expected to increase the flow to the glomerulus.
vasa recta, constitute the major resistance of the renal vas- The wall of the afferent arteriole, however, responds to
cular bed, ensuring high pressures within the glomeruli. stretch by contraction, thus reducing the diameter of the ar-
As noted already, the glomerular filtration rate depends teriole and therefore increasing the resistance to flow. This
largely on the net filtration pressure and the permeability of myogenic mechanism thus reduces variations in flow to the
Bowman's capsule. The net filtration pressure depends on glomerulus in the face of oscillations in blood pressure. Sec-
the blood pressure (glomerular capillary pressure), the in- ond, cells in the juxtaglomerular apparatus (JGA),which
tracapsular pressure, and the colloid osmotic pressure of is located where the distal tubule passes close to the Bow-
the blood plasma (see Table 14-9). Under normal condi- man's capsule between the afferent and efferent arterioles,
tions, the colloid osmotic pressure and intracapsular pres- secrete substances that modulate renal blood flow.
sure do not vary. The colloid osmotic pressure of plasma The juxtaglomerular apparatus is composed of three
can be elevated during dehydration, and the intracapsular types of cells (Figure 14-20):
pressure can be increased by the presence of kidney stones
obstructing the renal tubules; in both cases glomerular fil- Modified distal-tubule cells, which form the macula
tration rate will be reduced. In contrast, the seepage of densa and may monitor the osmolarity and flow of fluid
plasma through burned skin can lower the colloid osmotic in the distal tubule
pressure of plasma, which in turn could increase the
Specialized vascular cells, called granular cells, located
glomerular filtration rate. These examples, howevel; are ex-
between the afferent and efferent arterioles
ceptions rather than the rule.
Although blood pressure and cardiac output normally Secretory juxtaglomerular cells, modified smooth-mus-
increase during exercise, these changes have little effect on cle cells that are located primarily in the wall of the af-
the glomerular filtration rate in mammals because of regu- ferent arteriole
latory processes that control blood flow to the kidney. This
regulation is achieved by modulating the resistance to flow Under certain conditions, the juxtaglomerular cells release
in the afferent arteriole leading to each nephron and de- the hormone renin, which indirectly affects blood pressure
~4c;culadensa ~ f f e r e n arteriole
t
Figure 14-20 The juxtamedullary apparatus plays a key role in control- the macula densa; secretory juxtaglomerular cells in the wall of the affer-
ling blood flow through the glomerulus. This structure is composed of ent arteriole; and granular cells. [Adapted from Sherwood, 1993.1
several cell types including modified distal-tubule cells, which constitute
594 INTEGRATION OF PHYSIOLOGICAL SYSTEMS
.........................................
and, therefore, renal blood flow as described below. The absorption of various metabolites, ions, and water. The hu-
juxtaglomerular apparatus also releases various substances man kidneys produce about 180 liters of filtrate per day,
that act in a paracrine fashion to cause vasoconstriction or but the final volume of urine is only about 1 liter. Thus,
vasodilation of the afferent arteriole in response to in- over 99% of the water is reabsorbed. Of the 1800 g of
creased or decreased flow, respectively, through the distal NaCl typically occurring in the original filtrate, only 10 g
tubule. Thus myogenic and i ~ x t a ~ l o m e r u l afeedback-
r (or less than 1%) appear in the urine of persons consuming
control mechanisms work together to autoregulate the 10 g of NaCl per day. Varying amounts of many other fil-
glomerular filtration rate over a wide range of blood tered solutes are also reabsorbed from the tubular lumen.
pressures. In addition, some substances are secreted into the tubular
In addition to these autoregulatory mechanisms, the fluid. The renal clearance of a substance is a measure of the
glomerular filtration rate is subject to extrinsic neural con- extent to which it is reabsorbed or secreted in the kidneys,
trol. The afferent arterioles are innervated by the sympa- as explained in Spotlight 14-1.
thetic nervous system. Sympathetic activation causes vaso- To understand the relationship between clearance and
contriction of afferent arterioles and a reduction in reabsorption, let's consider glucose. A healthy mammal ex-
glomerular filtration. This response, which overrides any hibits a plasma glucose clearance of 0 ml smin-'. That is,
autoregulation, occurs when there is a sharp drop in blood even though the glucose molecule is small and is freely fil-
pressure, for example, as a result of extensive blood loss. tered by the glomerulus, normally it is completely reab-
The reduction in filtration rate helps to restore blood vol- sorbed by the epithelium of the renal tubule (Figure 14-21).
ume and pressure to normal. Conversely, an elevation in Glucose is fully reabsorbed because its loss in the urine
blood pressure reduces sympathetic vasoconstriction and would mean a loss of chemical energy to the organism.
enhances glomerular filtration, decreasing blood pressure Usually glucose appears in the urine only when the glucose
and volume. concentration in the blood plasma, and hence in the
Sympathetic activation can also cause contractions of glomerular filtrate, is very high. Figure 14-21 reveals
elements within the glomerulus, closing off portions of the that there is a maximum rate (milligrams per minute) at
filtering capillaries and effectively reducing the area avail- which glucose can be removed from the tubular urine by
able for filtration. The podocytes also are contractile, and reabsorption. This transfer maximum, or Tm, is about
when they contract, the number of filtration slits decreases. .
320 mg min-' in humans. Below plasma glucose levels of
Thus contraction of either or both of these elements can ef- about 1.8 mg-ml-', all the glucose appearing in the
fectively reduce the hydraulic permeability of Bowman's glomerular filtrate is reabsorbed. At about 3.0 mg-mlkl,
capsule. In the past, the hydraulic permeability of the the carrier mechanism is fully saturated, so that any addi-
glomerular membrane was thought to change only in dis- tional amount of glucose appearing in the filtrate will be
ease states that caused the membrane to become leaky. It passed out in the urine. The arterial plasma glucose con-
is now apparent that normal regulation of the glomerular centration in humans is normally held at about 1mg mlkl .
filtration rate can involve changes in the hydraulic perme- by an endocrine feedback loop involving insulin. Since this
ability of the glomerular membrane. level is well below the Tm for glucose, normal urine con-
A reduction in renal blood pressure, a fall in solute de- tains essentially no glucose. Because the high plasma glu-
livery to the distal tubule, andlor activation of the sympa-
thetic innervation induces release of the hormone renin
from the secretory juxtaglomerular cells located in the wall
/
of the afferent arteriole that carries blood into the glomeru- /
lar capillaries in Bowman's capsule. Renin is a proteolytic Filtered / /
enzyme whose release leads to increased levels of an-
giotensin I1 in the blood. This hormone has several actions,
one of which is t o cause general vasoconstriction (constric-
tion of arterioles), which raises the blood pressure, thereby
increasing both renal blood flow and the rate of glomeru- Reabsorbed
lar filtration. Angiotensin I1 may also cause constriction of Excreted
the efferent arterioles, raising glomerular blood pressure
and increasing filtration. Angiotensin I1 also stimulates re-
lease of the steroid aldosterone from the adrenal cortex and
vasopressin from the posterior pituitary. The role of these Plasma glucose (mg %)
hormones in promoting the tubular reabsorption of salts
and water is discussed later. Figure 14-21 The concentrat~onof glucose in the glomerular filtrate
(broken line) is proportional to the plasma glucose concentration. The re-
nal tubules are capable of reabsorbing the glucose by active transport
Tubular reabsorption (colored line) at rates up to 320 mg.min-' (Trn,). Glucose entering the
As the glomerular filtrate makes its way through the filtrate in excess of this rate is necessarily excreted via the urine (black
nephron, its original composition is quickly modified by re- line).
IONIC A N D OSMOTIC BALANCE 595
...............................................................................
SPOTLIGHT 14-1 In thls speclal case, the substance used, ~nulln,1s freely flltered
and unchanged by tubular absorption or secretion Therefore,
RENAL CLEARANCE the GFR and the clearance, C, of the substance are equal. sub-
stltutlng Cfor GFR glves, for lnulln,
The renal clearance of a plasma-borne substance 1s the volume
of blood plasma from whlch that substance 1s "cleared" (I e., vu
-- - I
completely removed) per unlt tlme by the kldneys. A substance CP
that IS freely flltered Into the nephron along wlth water, but that
IS nerther reabsorbed nor secreted in the kidney, permits the cal- SO the renal clearance IS glven by
culatlon of the glomerular filtration rate (GFR) merely by dlvld-
cose levels typical of diabetes mellitus exceed the reabsorp- oped by Alfred Richards and his coworkers in the 1920s. A
tion ability of the renal tubule, diabetics commonly have glass capillary micropipette is used to remove a minute
glucose in their urine. sample of the tubular fluid from the lumen of the nephron.
The details of tubular function vary from species to The osmolarity of the sample (expressed as milliosmoles
species. Our knowledge of the changes in urinary compo- per liter1 is then determined by measuring its freezing point.
sition along different portions of the nephron is based to a The lower the freezing point, the higher its osmolarity.
large extent on the technique of micropuncture, first devel- The stopped-flow perfusion technique, a modification of
......................................
Richards' original technique, can be used to isolate a por- The result is a tubular fluid that is isosmotic with respect to
tion of the lumen and analyze its action on injected samples the plasma and interstitial fluids. Stopped-flow perfusion
of a defined solution in vitro (Figure 14-22). experiments revealed that when the NaCl concentration in-
Microchemicalmethods are now used to determine the side the tubule is decreased, the movement of water also de-
concentrations of individual ion species in the sample. In a creases. This result is just the opposite of what would be ex-
technique developed more recently, a given segment of re- pected if the outward movement of reabsorbed water
nal tubule is dissected from the kidney and perfused in vitro occurred by simple osmotic diffusion, and it indicates that
with a defined test solution; analysis of the perfusate pro- water transport is coupled to active sodium transport (see
vides insight to the movement of substances across the iso- Chapter 4). The actual pumping of Naf takes place at the
lated tubule segment (Figure 14-23).The results of numer- basolateral (serosal) surface of the epithelial cells of the
ous studies using these techniques have detailed the roles of proximal tubule, just as it does in frog skin and gallbladder
various portions of the nephron in the reabsorption of salts epithelia. In amphibians, this active transport leaves the
and water, which are summarized in Figure 14-24. tubular lumen about 20 mV negative relative to the fluid
The proximal tubule, which initiates the process of con- surrounding the nephron. This potential difference proba-
centrating the glomerular filtrate, is most important in ac- bly accounts for the passive net diffusion of chloride out of
tive reabsorption of salts. In this segment, about 70% of the the proximal tubule as the counterion for sodium. In the
Na+ is removed from the lumen by active transport, and a proximal tubule, NaHCO, is the major solute reabsorbed
nearly proportional amount of water and certain other proximally and NaCl the major solute reabsorbed distally.
solutes, such as C1-, follow passively. So about 75% of the At the most distal portion of the proximal tubule
L
filtrate is reabsorbed before it reaches the loop of Henle. (where it joins the thin descending limb of the loop of
;"' Henle), the glomerular filtrate is already reduced to one
n:
fourth of its original volume. As a result of the reduction
in the volume of tubular fluid, substances that are not
actively transported across the tubule or that do not pas-
Pipette sively diffuse across it are four times as concentrated to-
Proximal tubule ward the end of the proximal tubule than in the original fil-
Bowman's capsule
trate. In spite of this great reduction in the volume of
tubular fluid, the fluid at this point is isosmotic relative to
1 the fluid outside the nephron, having an osmolarity of
about 300 mosm. L-l. It is interesting to note that the ac-
tive transport of NaCl alone can account for the major
changes in volume of the fluid along the proximal tubule
and for the increased concentration of urea and many other
filtered substances.
The proximal tubule is ideally structured for the mas-
sive reabsorption of salt and water. Numerous microvilli at
the luminal border of the tubular epithelial cells form the
so-called brush border (see Figure 14-15). These projec-
tions greatly increase the absorptive surface area of the
membrane, thereby promoting diffusion of salt and water
from the tubular lumen into the epithelial cell.
Glucose and amino acids are also reabsorbed in the
proximal tubule by a sodium-dependent mechanism and
are not normally present in the ultrafiltrate beyond the
Bowman's proximal tubule. Carriers on the apical membrane co-
capsule
transport sodium and glucose or amino acids from the ul-
Figure 14-22 The stopped-flow perfusiontechnique is used to study the trafiltrate into the cell. The uptake process, which is uphill
. function of various portions of the renal tubule in vitro. A micropipette is for glucose and amino acids, depends on the sodium elec-
inserted into Bowman's capsule (I), and oil (color)then is injected until it
trochemical gradient created by the Na+/K+ATPase in the
enters the proximaltubule. Perfusionfluid (gray) is injected through a sec-
ond pipette into the middle of the column of oil, forcing a droplet ahead basolateral membrane of the tubular cell. Once in the tubu-
of it (2). The tubule is full when the droplet reaches the far end of the lar cell, glucose and amino acids diffuse into the blood.
tubule (3).After about 20 minutes, the perfusion fluid is collected by the Phosphates, calcium ions, and other electrolytes nor-
injection of a second liquid behind the oil remaining nearthe glomerulus mally found in the blood are reabsorbed up to that required
(4). The ability of the tubule segment to reabsorb or secrete substances
by the body, and any excess is excreted. Parathyroid hor-
can be determ~nedby comparing the composition of the perfusate
before and after it is injected. [From "Pumps in the Living Cell" by mone modulates the reabsorption of phosphates and cal-
Arthur K. Solomon. Copyright 0 1962 by Scientific American, Inc. All cium by the kidney. Parathyroid hormone stimulates kid-
rights reserved.] ney la,25-hydroxylase activity, which in turn stimulates
I O N I C A N D OSMOTIC BALANCE 597
.............................
Perfusing Fluxes Figure 14-23 Perfusionof a dissected segment of re-
nal tubule and chemical analysis of the perfusate per-
mits determination of the fluxes of ions across the
tubular wall in vitro.
the production of calcitriol, the active form of vitamin D. in the urine-concentrating system of the nephron. The thin
Calcitriol released into the blood stimulates calcium reab- segment of the ascending limb has also been shown by per-
sorption and phosphate excretion from the kidney, as well fusion experiments to be inactive in salt transport, although
as calcium absorption from the gut and release from bone it is highly permeable to NaCl. Its permeability to urea is
(see Figure 9-29). low, and to water, very low. Again, this differing perme-
'
The descending limb and thin segment of the ascending ability plays a key role in the urine-concentrating mecha-
limb of the loop of Henle are made up of very thin cells nism of the nephron.
containing few mitochondria and no brush border. In vitro The medullary thick ascending limb differs from the
perfusion studies have demonstrated that there is no active rest of the loop of Henle in that it exhibits active transport
salt transport in the descending limb. Moreover, this seg- of NaCl outward from the lumen to the interstitial space
ment exhibits very low permeability to NaCl and low per- (see Figure 14-24).This portion, along with the rest of the
meability to urea but is permeable to water. As discussed ascending limb, has a very low permeability to water. As a
later, this differential permeability plays an important role result of NaCl reabsorption, the fluid reaching the distal
Distal Figure 14-24 The movement of ions, water, and other substances
tubule in and out of the filtrate along the renal tubule determines the
Bowman's
capsule
'
Proximal + /+
- 1 composition of the urine. In this schematic diagram, the fluxes of
NaCI, water, and urea are shown in different portions of the rnam-
malian renal tubule. The numbers indicate the flltrate tonicity in
milliosrnolesper liter. The relative rates of active transport of NaCl
are indicated by the size of the arrow. The permeability of the stip-
pled portion of the collecting duct IS regulated by
antidiuretic hormone (ADH).[Adapted from Pitts, 1959.1
Cortex
400
Outer
medulla
duct
Inner
medulla
Loop
o f Henle 1200
-, Active transport
of NaCl
7 Passive diffusion
of urea
II)Passive diffusion
of H,O
Pass~ved ~ f f u s ~ o n
of NaCl
598 INTEGRATION OF PHYSIOLOGICAL SYSTEMS
.........................................
tubule is somewhat llyposmotic relative to the interstitial Tubular lumen
fluid. The importance of salt reabsorption by the thick as-
cending tubule is discussed later in the section on the urine-
A
concentrating mechanism.
Na+
The movement of salt and water across the distal tubule
is complex. The distal tubule is important in the transport Glucose
of K+, H+,and NH3 into the lumen and of Na-, C1-, and
HC0,- out of the lumen and back into the interstitial fluid.
As salts are pumped out of the tubule, water follows pas-
sively. The transport of salts in the distal tubule is under
endocrine control, and is adjusted in response to osmotic
conditions.
Because the collecting duct is permeable to water, wa-
ter flows from the dilute urine in the duct into the more
concentrated interstitial fluid of the renal medulla (see
Figure 14-24).This is the final step in the production of a
hyperosmotic urine. The water permeability of the duct is
variable and controlled by antidiuretic hormone (ADH).
Thus the rate at which water is absorbed is under delicate
feedback control. The collecting duct reabsorbs NaCl by
active transport of sodium. The inner medullary segment of
the collecting duct, toward its distal end, is highly perme-
able to urea. The significance of this will become clear in
our later discussion of the countercurrent mechanism that
concentrates the urine in the collecting duct. Figure 14-25 Several transport systems are involved in reabsorption of
Now that we have summarized the movement of watel; Na+ in the proximaltubule and ascending limb ofthe loop of Henle in the
ions, and glucose out of the tubular filtrate, let's exam- mammalian kidney. (A) Sodium passively crosses the apical membrane
ine sodium reabsorption in the nephron more closely. In via Na/ZCI/K and glucose/Na+ cotransporters. A Na7/K+ATPase in the
basolateral membrane actively removes Na+from the cell into the blood;
the proximal tubule and ascending limb of the loop of
K+ and CI- exit via ion channels down their concentration gradient. (B)
Henle, sodium is transferred across the apical membrane of The movement of Na+ down its electrochem~calgradient into the cell
the tubular epithelium via cotransporters and then is ac- also energizes the outward movement of protons via an electrically neu-
tively transported into the blood via a Na+/K+ ATPase tral Na+/H+exchanger. CO, in the blood diffuses into the cell, where car-
(Figure 14-25A). The electrochemical gradient for Nai be- bonic anhydrase (c.a.) ensures a high rate of proton delivery to the ex-
changer. A basolateral sodium pump transports Na* from the cell into the
tween the ultrafiltrate and the blood favors the diffusion of
blood. K+ and HCO, exit vla ion channels down their electrochemical
Na+ through channels in the apical membrane from the ul- gradient.
trafiltrate into the tubular cells. Sodium is also exchanged
for a proton via an electrically neutral Na+/H+exchanger;
in this case, the downhill movement of Na+ energizes the Figure 14-26A, renin acts to cause a rise in blood levels of
uphill movement of H+into the lumen (Figure 14-25B). Far- angiotensin I1 and subsequently aldosterone; the latter pro-
ther along in the distal tubule and collecting duct, sodium motes sodium reabsorption from the filtrate.
reabsorption is coupled t o secretion of protons into the Renin, a proteolytic enzyme, cleaves angiotensinogen,
urine by acid-secreting cells, which are involved primarily a glycoprotein molecule that is manufactured in the liver
in p H regulation. These cells are described in detail later. and is present in the a,-globulin fraction of plasma pro-
Sodium chloride represents more than 90% of the 0s- teins. Cleavage of angiotensinogin releases a 10-residue
motic activity of the extracellular fluid. Because reabsorp- peptide, angiotensin I. Angiotensinogen-converting enzyme
tion of salt results in the reabsorption of water, the amount (ACE) then removes two additional amino acids to form
of salt in the body is an important determinant of the vol- the 8-residue peptide, angiotensin I1 (Figure 14-26B). Much
ume of the extracellular fluid (ECF).If ECF volume is large, of the formation of angiotensin I1 occurs during the passage
then blood pressure tends to rise. The converse is true for of blood through the lungs. Angiotensin I1 stimulates the
a reduction in ECF, for example, as a result of blood loss. secretion of aldosterone from the adrenal cortex and also
Thus blood pressure is an indication of blood volume, causes a general vasoconstriction, which raises blood pres-
which in turn is a reflection of the salt content of the body. sure. Removal of the amino-terminal aspartic acid residue
When cells of the macula densa portion of the juxta- from angiotensin II yields angiotensin 111, which also causes
glomerular apparatus sense a decrease in blood pressure secretion of aldosterone from the adrenal cortex but to a
andlor solute delivery to the distal tubule, they stimulate lesser extent than angiotensin 11.
release of renin from the juxtaglomerular cells in the walls Like other steroid hormones, aldosterone diffuses
of the afferent arteriole (see Figure 14-20). As outlined in across the cell membrane and binds to cytoplasmic recep-
I O N I C A N D OSMOTIC BALANCE 599
.............................
A Figure 14-26 The renin-angiotensin
Aldosterone system plays an important role in control-
\ ling sodium reabsorption in the mam-
Increased N a f malian kidney. (A) Renin is liberated by
reabsorptlon secretory cells in the juxtaglomerular
apparatus (JGA) in responseto decreased
pressure in the afferent arteriole and to
& Adrenal
tubule
low Na+ concentration in the distal tubule.
Circulating renin leads to an increase
in the titer of angiotensin II and aldos-
terone. Aldosterone stimulates Na+ reab-
sorption from the filtrate in the renal
tubule. (B) Renin is a proteolytic enzyme
that cleaves angiotensinogen, an a2-glob-
.... ulin, yielding angiotensin I. Another pro-
teolytic enzyme' then removes the two
I General systemic
vasoconstriction
carboxyl-terminal residues to give angio-
tensin II.
Angiotensin II /\,
Converting
enzyme
\
Angiotens~nogen Asp-Arg-Val-Tyr-lle-His-Pro-Phe-His-Leu-Leu-Val-Tyr-Ser-Protein
(Renin Substrate)
Angiotensin l
1Renin
Asp-Arg-Val-Tyr-lle-His-Pro-Phe-His-Leu
Angiotensin II
1Converting enzyme
Asp-Arg-Val-Tyr-lle-His-Pro-Phe
tors in target cells, leading to an increase in transcription of 2. Metabolic hypothesis: Increase in the production of
specific genes and ultimately synthesis of the encoded pro- ATP to power the sodium pump, perhaps due to al-
teins (see Figure 9-9). Aldosterone acts on the cells of the dosterone-stimulated increase in fatty acid metabolism.
tubular epithelium to increase sodium reabsorption but 3. Permease hypothesis: Increased permeability of the
does so without affecting water permeability. Three mech- apical membrane to Na+ ions, presumably due to an
anisms have been proposed to account for the aldosterone- increase in the number of sodium channels in the
induced increase in sodium reabsorption across tubular ep- membrane.
ithelial cells (Figure 14-27):
Quite possibly, all three mechanisms operate in tubular cells
1. Sodium-pump hypothesis: Increased activity of stimulated by aldosterone.
Na+/K+ATPase in the basolateral membrane, perhaps Increased circulating levels of angiotensin I1 also in-
due to changes in membrane structure that enhance crease the synthesis of vasopressin, also called antidiuretic
ATPase activity as well as to increased synthesis of the hormone (ADH),in the hypothalamus and its release from
pump protein. the posterior pituitary (see Figures 9-5 and 9-7). Vaso-
Tubular secretion
The nephron has several distinct systems that secrete sub-
stances by transporting them from the plasma into the
tubular lumen. Most thoroughly investigated are the sys-
tems for secretion of K+, H+,NH3,organic acids, and or-
ganic bases. Although the number of secretory mechanisms
and transport molecules must be limited, nevertheless, the
nephron is capable of secreting innumerable "new" sub-
stances, including drugs and toxins, as well as endogenous,
naturally occurring molecules. How is the nephron able to
recognize and transport all these diverse substances? The
answer seems to reside in the role of the vertebrate liver in
modifying many such molecules so that they can react with
the transport systems located in the wall of the nephron.
These secretory mechanisms are important because they re-
move potentially dangerous substances from the blood. In -
'
the liver, many of these substances, along with normal
metabolites, are conjugated with glucuronic acid or its sul-
fate. Both these classes of conjugated molecules are actively
transported by the system that recognizes and secretes or-
ganic acids. Since they are highly polar, these conjugated
molecules, once deposited by the transport machinery in
the lumen of the nephron, cannot readily diffuse back
across the wall of the nephron into the peritubular space
and from there into the blood, and so these substances are
excreted in the urine.
Figure 14-27 Aldosterone, a steroid hormone that stimulates gene ex-
pression, increases sodium reabsorption in the kidney. Depicted here are
three mechanisms proposed t o explain the effect of aldosterone: an in-
crease in the activity of the Na+/K+ ATPase directly (sodium-pump hy-
pothesis) or indirectly by increasing ATP levels (metabolic hypothesis),
and an increase in the activity of sodium channels (permease hypothesis).
[Adapted from M. E. Hadley, 1992.1
solateral membrane to pump protons into the blood, accompanied by often fall; as a result, bicarbonate levels in the filtrate are re-
the reabsorption of CIk. Both cell types contain carbonic anhydrase (c.a.),
duced and less is available to act as a buffer. Under such
which rapidly forms Ht and HCO, ions from C02diffusing into the cell
from the blood. conditions, ammonia is a major vehicle for elimination of
excess acid. Ammonia is produced within the renal tubular
cells by enzymatic deamination of amino acids, especially
the extracellular flood. The basolateral membrane of the glutamine (see Figure 14-30). In its nonpolar, un-ionized
A-type cell also contains K+ channels, and K + is cycled
through this membrane by the Na+/K+ATPase. Thus acid-
Tubular lumen Blood
ification of the filtrate by A-type cells is coupled to sodium
reabsorption.
The distal tubule and collecting duct also contain base- Glutamine
secreting cells, called B-type cells. These cells have a chlo-
ridelbicarbonate exchanger in the apical membrane; this
exchanger differs from the band 3-type protein found in
the basal membrane of the A-type cell. As illustrated in
Figure 14-29B, B-type cells contain carbonic anhydrase and
secrete bicarbonate into the lumen of the tubule in ex-
change for chloride. Protons and chloride ions move across
the basolateral membrane via a proton ATPase and chlo-
ride channels. Basolateral
membrane
A mammal can regulate its body pH by altering the
activity of these A-type and B-type cells. The activity of Figure 14-30 Buffering of the renal filtrate by HPO, and NH,' permits
A-type cells and, therefore, acid secretion increase during greater secretion of protons. The phosphate ions in the lumen arise by fil-
acidosis, whereas increased B-type cell activity and bicar- tration, whereas the ammonium ions arise by passive diffusion of NH,
bonate secretion are associated with alkalosis. Changes in from the blood across the tubular cells or by intracellular breakdown of
glutamine. Glutamine (and other amino acids) enter the tubular cells via
the activity of the A-type cells involve alteration in both the
basolateral transporters and is deaminated, yield~ngNH,, which diffuses
proton ATPase activity in the apical membrane and the across the apical membrane into the lumen. Because the membrane is
number of bicarbonatelchloride exchangers present in the largely impermeable to both H,P04- and NH4+,both ions are trapped in
basal membrane. the urine and excreted.
form, ammonia freely diffuses across the cell membrane I 1 Medulla I
into the lumen where it reacts with protons forming NH,+
ions. Because the highly polar NH4+is impermeant, it traps
both nitrogen atoms and protons in the urine, thus serving
I lJ---i
'Ortex Outer zone
Urine-Concentrating Mechanism
The urine of birds and mammals becomes concentrated by
osmotic removal of water from the filtrate in the collecting
ducts as they course through the renal medulla. There is a
clear-cut correlation between the architecture of the verte-
brate kidney and its ability to manufacture a urine that is
hypertonic relative to the body fluids. Kidneys capable of
producing a hypertonic urine (i.e., those of mammals and
birds) all have nephrons featuring the loop of Henle. More-
over, the ability of a mammal to concentrate the urine is di- Inner medulla
rectly related to the length of the loops of Henle in its kid-
neys. The loops of Henle are longest in desert dwellers, such
as the kangaroo rat; these longer loops produce larger over- /
Outer medulla
all gradients in osmolarity from renal cortex to medulla,
thus permitting more efficient osmotic extraction of water Figure 14-31 Solute concentrations in the interstitiurnof the mammalian
from the collecting duct. In general, the longer the loop and kidney progressively increase from the cortex into the depths of the
the deeper it extends into the renal medulla, the greater the medulla. Shown here are the interstitial concentrations(in millimoles per
concentrating power of the nephron. Thus, desert mam- liter) of urea, sodium, and chloride at different depths Note that most of
the increase in urea concentration occurs across the inner medulla,
mals have both the longest loops of Henle and the most hy-
whereas most of the increase in NaCl concentration occurs across the
pertonic urine. outer medulla. Since the osmotic contributions of Na+ and CI- sum, the
In addition to this correlation between the anatomy total osmotic contributions of NaCl and urea are about equal deep within
and concentrating ability of the nephron, the tonicity of the the medulla. [Adapted from Ulrich et al., 1961.1
interstitial fluid progressively increases toward the deeper
regions of the renal medulla (Figure 14-31)for reasons dis-
cussed later. These findings led B. Hargitay and Werner tubule is isosmotic with respect to the extracellular fluid
Kuhn to propose in 1951 that the loop of Henle acts as a at that point (i.e., the outer portion of the renal medulla),
countercurrent multiplier (Spotlight 14-2).Though a very having a concentration of about 300 mosm-L-' (see
attractive and plausible hypothesis, it was initially hard to Figure 14-24).The concentration of the fluid gradually in-
test because of the difficulty of sampling the intratubular creases as it makes its way down the descending limb to-
fluid in the thin loop of Henle. Determinations of the melt- ward the hairpin turn in the loop, where its concentration
reaches 1000-3000 mosm. L-l in most mammals. At this .
ing point of the fluid in slices of frozen kidney and subse-
quently in situ perfusion experiments of segments of the point, too, it is nearly isosmotic relative to the surrounding
loop provided experimental support for the countercurrent extracellular fluid in the deep portion of the renal medulla.
hypothesis. This increase in the osmolarity of the tubular fluid flowing
These studies showed that the fluid entering the de- down the descending limb occurs because the wall of the
scending limb of the loop of Henle from the proximal descending limb is relatively permeable to water, but far less
604 INTEGRATION OF PHYSIOLOGICAL SYSTEMS
.......................................
SPOTLIGHT 14-2 gressively higher. As the fluid rounds the bend and begins flow-
ing out the other limb, its salt concentration progressivelyfalls as
COUNTERCURRENT a result of the cumulative effect of outward NaCl transport along
the length of the outflow limb. By the time it reaches the end of
, SYSTEMS that limb, its osmolarity is slightly lower than that of the fresh fluid
beginning its inward flow in the other limb. This establishes a salt
In 1944 Lyman C. Craig published a method of concentrating gradient along the tube.
chemical compounds based on the countercurrent principle. This example resemblesthe loop of Henle in principle but not
This method has proved useful in many industrial and laboratory in detail. The loop of Henle has no common wall dividing the two
applications. As in many other instances, human ingenuity turns limbs; nevertheless, the limbs are coupledfunctionally through the
out to be a reflection of Nature's inventiveness; countercurrent interstitial fluid, so that the NaCl pumped out of the ascending
mechanisms have been found to operate in a variety of biologi- limb can diffuse the short distance toward the descending limb
cal systems, including the vertebrate kidney, the gas-secreting and cause osmotic reabsorption of water from that limb. Several
organ of swim bladders and gills of fishes, and the limbs of vari- important points should be noted about countercurrent systems
ous birds and mammals that live in cold climates. such as the loop of Henle and the simple model illustrated here.
The principle can be illustrated with a hypothetical counter- First, the standing concentration gradient set up in both
current multiplier that employs an active-transport mechanism limbs is due to both the continual movement of fluid through the
much like the one that operates in the mammalian kidney. The system and the cumulative effect of transfer from the outflow limb
model shown in part A of the accompanyingfigure consists of a to the inflow limb. The gradient would disappear if either fluid
tube bent into a loop with a common dividing wall between the movement or transport across the membrane were to cease.
two limbs. A NaCl solution flows into one limb of the tube and Second, the difference in concentrationfrom one end to the
then out the other. Let us assume that, within the common wall other of each two limbs of the countercurrent multiplier is far
separating the two limbs of the tube, there is a mechanism that greater than the difference across the part~tionseparating the
actively transports NaCl from the outflow limb to the inflow limb limbs at any one point (part B of the figure). As a consequence,
of the tube, without any accompanying movement of water. As the countercurrent multiplier can produce greater concentration
bulk flow carries the fluid along the inflow limb, the effect of NaCl changes than would be attained by a simple transport epithe-
transport is cumulative, and the salt concentration becomes pro- lium without the configuration of a countercurrent system. The
longer the multiplier, the greater the concentration differences
that can be attained.
A
Increasing salt gradient Third, the multiplier system can work only if it contains an
b
Bulk - asymmetry. In the model in part A, there is an active energy-
flow requiring net transport of NaCl in one direction across the par-
tition. A passive countercurrent system, such as one used to con-
serve heat, does not requlre the expenditure of energy (part C of
the figure). In the extremities of birds and mammals that inhabit
cold climates, fgr example, a temperature differential exists be-
tween the arterial and venous flow of blood, because the blood
is cooled as it descends into the leg. As a result of this asymme-
try and the countercurrent arrangement of the vessels, the arte-
rial blood gives up some of its heat to the venous blood leaving
the leg, thereby reducing the amount lost to the environment.
Collecting
duct
Neurosecretory
termrnals rn
pltultary
Hlgh
plasma
absorptlon
H,O perm
608 I N T E G R A T I O N O F P H Y S I O L O G I C A L SYSTEMS
.........................................
mones are applied to frog skin and toad bladder, they in- nism for the production of a filtrate. These fish are hypo-
crease the water permeability of those epithelia. tonic to their environment and so lose water continually
To summarize the mechanisms we've discussed, the for- across the skin and gills. Their problem is water conserva-
mation of urine in the mammalian kidney begins with the tion and they produce only small volumes of urine. Little
concentration of the glomerular filtrate into a hyperosmotic urea is formed and ammonia is excreted across the gills.
fluid in the proximal tubule. About 75% of the salt and wa- Amphibians and reptiles appear incapable of produc-
ter are removed from the filtrate in osmotically equivalent ing a hypertonic urine (i.e., of higher osmolarity than the
amounts as it passes through the proximal tubule, leaving plasma), because they lack the countercurrent system of the
urea and certain other substances behind. As the filtrate loop of Henle that is necessary to produce urine of signifi-
moves through the loop of Henle and the distal tubule, cantly greater osmolarity than the plasma. Only mammals
there is little net change in its osmolarity, but the counter- and birds are known to have a renal countercurrent orga-
current mechanism sets up a standing concentration gra- nization, and thus only these animals, apparently, have
dient in the medullary interstitium along the length of the their plumbing so organized as to allow osmotic counter-
loop. This gradient provides the basis for the osmotic re- current multiplication. The avian kidney contains a mix-
moval of water from the urine as it makes its way down the ture of reptilian-type and mammalian-type nephrons. That
collecting duct within the medulla. Interestingly, this is, some avian nephrons lack a loop of Henle, and in some
process takes place without active transport of water at any birds the loop is oriented perpendicular to the collecting
place along the nephron. duct, producing a less efficient concentrating mechanism.
An animal can experience osmotic stress owing to The elasmobranch Raja erinacea (a skate) has been
changes in temperature or salinity and owing to the inges- shown to have a complex renal tubule organization that
tion of food and drink. Perturbations in the osmotic state has the anatomic requisites for countercurrent multiplica-
of the body fluids are minimized through feedback mech- tion. However, the skate nephron is functionally quite dif-
anisms by which the osmoregulatory organs adjust their ac- ferent from the mammalian nephron. As we have seen, the
tivity so as to maintain the internal status quo. These feed- mammalian kidney excretes urea and retains water to pro-
back control mechanisms may be neural, endocrine, or a duce a hypertonic urine. The elasmobranch kidney, in con-
combination of the two. In mammals, adjustments in the trast, retains urea (which is used as an osmolyte) and does
volume and concentration of the urine are the primary not produce a concentrated urine. Tubular bundles consti-
means for maintaining osmotic homeostasis. In response tute the countercurrent system in elasmobranch kidneys.
to osmotic stress and other signals, mammals can regulate These tubular bundles have been described in the kidneys
several aspects of urine formation including (1) the of marine elasmobranchs, which have high levels of urea in
glomerular filtration rate, (2) the rate at which salts and their tissues and reabsorb urea from the kidney ultrafiltrate.
water are absorbed from the lumen of the renal tubule, Freshwater stingrays, on the other hand, do not reabsorb
(3) secretion of unwanted substances, and (4)the rate at filtered urea and their kidneys lack tubular bundles, indi-
which water is osmotically drawn out of the pre-urine in cating that the bundles are the site of urea reabsorption.
the collecting duct. Thus, the function of the countercurrent organization of
the elasmobranch nephron may be to conserve urea.
NONMAMMALIAN VERTEBRATE
KIDNEYS EXTRARENAL OSMOREGULATORY
In the kidneys of the marine hagfishes (class Cyclostomata),
ORGANS IN VERTEBRATES
the nephrons possess glomeruli but no tubules, so the Bow- As indicated in the previous section, many vertebrates rely
man's capsules empty directly into collecting ducts. The on extrarenal osmoregulatory organs to maintain osmotic
kidneys are used largely to excrete divalent ions (e.g., Ca2+, homeostasis. First, we consider specialized glands for ex-
Mg2+,and S 0 4 2 - ) and carry out little or no osmoregula- creting salt found in some animals, and then see how fish
tion. Thus, the extracellular fluids of the most primitive liv- gills are used for osmoregulation.
ing vertebrate, the hagfish, are relatively similar to seawa-
ter in concentration of major salts, and their plasma is Salt Glands
essentially isotonic relative to seawater (see Table 14-1). Elasmobranchs, marine birds, and some reptiles possess
As a general rule, the kidneys in freshwater teleosts glands that secrete salt by cellular mechanisms similar to
have larger glomeruli and more of them than do those in sodium reabsorption in the mammalian kidney.
their marine relatives. Because their bodies are hypertonic
to the environment and water diffuses into their bodies, Elasmobranch rectal gland
freshwater teleosts maintain water balance by producing Marine elasmobranchs, although slightly hypertonic to sea-
large volumes of dilute urine. The kidney nephrons in cer- water, have a much lower NaCl content than seawater. As
tain marine teleosts have neither glomeruli nor Bowman's a result there is a continual influx of NaCl into the body of
capsules. In such aglomerular kidneys, the urine is formed the animal. The excess salt is removed largely by the rectal
entirely by secretion because there is no specialized mecha- gland, which produces a concentrated salt solution and is
I O N I C A N D OSMOTIC BALANCE 609
......................................
the major (perhaps the only) extrarenal site for secretion of port system also in the basolateral membrane. Thus as Naf
excess NaCl by marine elasmobranchs. The gland func- and K+ cycle across the basal membrane, the C1- level in-
tions to regulate extracellular volume by controlling the side the cell rises above that in the tubular lumen; eventu-
amount of NaCl in the body. ally C 1 exits via chloride channels in the apical (mucosal)
The rectal gland consists of a large number of blind- membrane moving down its concentration gradient. The
ending tubules that drain into a duct, which opens into the overall effect is the movement of C1- from the serosal
intestine near the rectum. The fluid produced by the gland (blood)side of the tubular wall into the lumen. This creates
can have a slightly higher salt concentration than seawater an electrical potential, with the serosal side positive and the
but is isosmotic to the plasma of the fish. The blood of elas- lumen negative; the resulting electrochemical gradient for
mobranchs is also slightly hyperosmotic to seawater but sodium permits diffusion of Nat from the serosal side
has a much lower salt concentration, the osmolarity of the through paracellular pathways into the lumen. Water fol-
blood being made up by high concentrations of urea and lows the transport of NaCl and is distributed passively
trimethylamine oxide (TMAO).Elasmobranchs are able to across the tubular wall, but the wall is impermeable to urea
tolerate high levels of urea, which normally causes the dis- and TMAO. Thus the rectal gland produces a solution that
sociation of multi-subunit enzymes, thus inhibiting their ac- has a much higher NaCl concentration than the blood but
tivity. In contrast, TMAO promotes the association of sub- is isosmotic with the blood.
units, thereby counteracting the effect of urea. Urea and The hearts of dogfish sharks contain a natriuretic pep-
TMAO do not appear in the rectal gland fluid, only NaCl. tide hormone that stimulates chloride secretion in perfused
Formation of the secreted fluid in the rectal gland does rectal glands. Although circulating natriuretic peptide lev-
not involve filtration of the blood; rather NaCl is secreted els have not yet been measured in elasmobranchs, it is pos-
into the lumen of the tubule and water follows. The cells of sible that natriuretic peptides released from the heart into
the tubule wall of the rectal gland consist of a single type of the circulation stimulate secretion by the rectal gland, re-
cell, a salt-secreting cell similar to the chloride cell found in ducing extracellular volume. The appropriate stimulus for
the gills of marine teleosts. This cell has a very extensive, the release of the natriuretic peptide would seem to be a rise
folded, basolateral membrane, whose surface area is much in venous pressure, that is, filling pressure of the heart. In
larger than that of the apical (mucosal) membrane. The ba- fact, the heart of a teleost fish, the rainbow trout, has been
solateral (serosal) membrane contains high concentrations shown to contain natriuretic -peptide
- hormone, which is re-
of a Na+/K+ATPase, which pumps Na+ out and K+ into leased into the circulation by increased venous pressure.
the cell; the K+, however, cycles back out through the many
potassium channels also present in the basolateral mem-
brane (Figure 14-36).The activity of the Na+/K+ATPase
generates a large sodium gradient across the basal mem-
brane, which drives NaCl uptake via a Na12CVK cotrans- EiI Do you think dinosaurs had salt glands?
Lumen of
Lobe Gland
Blood
flow
Figure 14-37 Marine birds malntain osmotic balance by excretion of a blood to the tubule, since the uphill gradient of salt concentration be-
concentrated salt solution from glands located above the orblt. (A) The tween cap~llaryand tubule lumen is thereby minimized at each point
avian salt gland consists of a longitudinal arrangement of many lobes, along the length of the tubule. (C)The secretory cells constituting the
which drain via a central canal into a duct that carries secretions to the tubularwall transport NaCl from the blood into the lumen via the mech-
nasal passages.(B) Each lobe consists of tubules and capillaries arranged anism depicted in Figure 14-%.These cells have a brush border and con-
radially around a central canal. Single tubules are surrounded by capil- tain many mitochondria. [Part A adapted from Schmidt-Nielsen, 1960;
laries In which blood flows counter to the flow of secretory fluid in the part B from "Salt Glands," by K. Schmidt-N~elsen.Copyright O 1959 by
tubule. This countercurrent flow facilitates the transfer of salt from the Scientific American, Inc. All rights reserved.]
secretion takes place across the epithelium of the secretory salt transport. Since this inhibitor does not pass across ep-
tubules, which consists of characteristic salt-secretingcells. ithelia and can block the pump only by direct contact with
These have a profusion of deep infoldings in the basolateral the ATPase, the sodium-transport mechanism appears to
membrane and are heavily laden with mitochondria. As in operate in the basal membrane of the epithelial cells, as it
many other transport epithelia, adjacent cells are tied to- does in the rectal gland. Increased salt secretion is associ-
gether by junctions, which preclude the massive leakage of ated with increased Na+/K+ ATPase activity in the salt
water past the cells, from one side of the epithelium to the gland. The Na+/K+ ATPase also occurs to some extent in
other. These cell junctions, however, are not as tight as the apical membrane of the bird nasal gland. The basal
those binding cells of the frog skin together but are leaky al- membrane of the salt-gland epithelium also contains a
lowing the paracellular movement of ions, as in the rectal Na12CVK cotransporter and potassium channels, and the
gland. apical membrane contain chloride channels. The net result
The formation of fluid in the nasal gland, as in the rec- is movement of NaCl from the blood across the epithelium
tal gland, does not include filtration of the blood. The ab- into the lumen of the salt gland (Figure 14-37C).
sence of filtration can be deduced from the failure of small As we saw earlier, the salt solution produced by the
filterable molecules (e.g., inulin or sucrose) that are injected elasmobranch rectal gland is isosmotic to plasma; in con-
into the bloodstream to appear in the gland fluid. High con- trast, the fluid produced by the nasal gland is hyperosmotic
centrations of a Na+/K+ATPase have been demonstrated to plasma. In both cases, the gland fluid has a high salt
in the basolateral membrane of the tubular cells. Applica- concentration, but the osmolarity of the blood of elasmo-
tion of ouabain to the basal surface of the epithelium blocks branchs is much higher than that of birds and reptiles. It is
I O N I C A N D OSMOTIC BALANCE 611
...............................................................................
not clear how the solution produced by the nasal glands of drinking saltwater is to reduce extracellular volume while
birds and reptiles is concentrated. It is possible that the ini- increasing NaCl levels in extracellular fluid and the blood
tial solution in the apex of the tubule is isosmotic to plasma (Figure 14-38A).The salt level in the gut thus will drop be-
and becomes more concentrated as it passes down the cause of salt loss to the body and diffusion of water from
tubule. The cells of the secretory epithelium of a single the body into the gut. After a while the osmolarity of the
tubule become larger, with deeper paracellular channels, to- gut fluids will fall below that of the body, so that water
wards the base of the tubule, indicating that the fluid may movement between the body into the gut will reverse, that
become more concentrated towards the base of the tubule. is, water moves into the body, following the movement of
Those birds that can produce the most concentrated salt so- salt and expanding extracellular volume. The initial reduc-
lutions have the largest secretory cells, with long paracellu- tion in extracellular volume inhibits nasal fluid production
lar channels between cells. In addition, the avian salt gland immediately after drinking seawater. The subsequent ele-
and its blood flow are organized as a countercurrent system vation of both extracellular volume and salt content, acts
and this might aid in concentration of the salt solution. The as a strong stimulus for salt secretion, thus there is often a
capillaries are so arranged that the flow of blood is paral- short delay between drinking saltwater and secretion from
lel to the secretory tubules and occurs in the direction op- the nasal gland. Since the solution secreted from the salt
posite to the flow of secretory fluid (see Figure 14-37B). gland is more concentrated than the seawater taken in, the
This flow maintains a minimal concentration gradient from bird ends up gaining osmotically free water, as illustrated in
blood to tubular lumen along the entire length of a tubule; Figure 14-38B.
it thereby minimizes the concentration gradient for uphill Regulation of the secretory activity of the avian
transport from the plasma to the secretory fluid. salt gland involves both parasympathetic neural control
The salt gland is not always active but responds to a salt and neuroendocrine control through the pituitary
load and/or expansion of the extracellular space. When (Figure 14-39). Osmoreceptors in the hypothalamus re-
birds drink seawater, water will diffuse from the body into spond to an increase in plasma tonicity by a sensory dis-
the gut because seawater has a higher osmolarity than the charge. This response, together with input from extracra-
body fluids. At the same time NaCl will diffuse from the nial osmoreceptors and/or volume receptors, activates
seawater in the gut into the body. Thus the initial effect of parasympathetic cholinergic neurons that innervate the salt
Neurohypophys~s
Dehydrat~on,
hypersalln~ty
Figure 14-39 The increase in avian salt-gland secretion in response t o leased from the heart in response t o low venous pressure, also directly
a rise in blood osmolarity and decrease in blood pressure is mediated by stimulates secretion. Pituitary secretion of ACTH in response to increas-
direct and indirect mechanisms.Stimulation of osmoticallysensitive neu- ing blood osmolarity indirectly promotes salt secretion by stimulating re-
rons in the hypothalamus and sensory input from peripheral osmorecep- lease of corticosterone (CS) from the adrenal cortex. This hormone acts
tors activate direct parasympathetic pathwaysto the salt gland and to the directly on the gland making it responsive to blood tonicity.
vessels supplying blood to the gland. Atrial natriuretic peptide (ANP), re-
GILL STRUCTURE
Gill Operculum
Figure 14-41 Fish gills function as both respiratory and osmoregulatory ter, Na+can move in and out of the blood in the larnellae. Black arrows
organs. These drawings show a portion of the teleost gill in increasing indicatewater flow; red arrow and dotted lines indicate blood flow.
magnification. In addition to gas exchange between the blood and wa-
614 I N T E G R A T I O N OF PHYSIOLOGICAL SYSTEMS
......................................
Figure 14-42 The eplthellum of the teleost gill 1s composed largely
of pavement cells Interspersed w ~ t ha few mucous and chlor~decells
(A) Drawlng of lamellar eplthellum showlng typlcal dlstrlbutlon of pave-
ment, mucous, and chlorlde cells The chlorlde cells tend t o be at the d
cells, especially near the pit that develops on the apical (mu- the case of marine teleosts, the secretion of salt occurs
cosal or external) border of these cells in fishes that have be- against an osmotic gradient and there is no movement of
come adapted to high salinities. water following the movement of salt. Thus the shark rec-
The mechanism of salt transport by chloride cells is tal gland, avian nasal gland, seawater teleost gill, and the
similar to that of the salt-secreting cells illustrated in thick ascending limb of the loop of Henle in the mam-
Figure 14-36. Thus, chloride cells have high levels of a malian kidney tubule all appear to contain salt-secreting
Na+/K+ATPase associated with Na/2Cl/K cotransporters cells that transport NaCl by the same basic mechanism (see
in the basolateral membrane and a chloride channel in the Figures 14-25A and 14-36). In the mammalian kidney,
apical membrane. Each chloride cell is associated with an however, the direction of salt transport is into the blood
accessory cell (distinct from a pavement cell), and Na+ dif- rather than into the environment, as in the other cases.
fuses from blood to seawater through the less-tight para- Since chloride cells were first characterized and associ-
cellular channel between the chloride and accessory cell. In ated with the transport of C1- across the gills of marine
I O N I C A N D O S M O T I C BALANCE 615
...............................................................................
teleosts, they have been found to also mediate the exchange TABLE 14-11
Physiological adaptation accompanying movement of fish to
of other ions including ca2+. F~~ instance, ca2+ in the wa-
water of differing salinity
ter is taken up via calcium channels in the apical membrane
of the chloride cells and then actively transported into the (A) Freshwater + saltwater
blood via a Ca2+ATPase, present at high levels in the ba- 1. Proton ATPase that powers active uptake of NaCl is down-
solateral membrane. regulated.
uptake across the apical membrane, whereas a Na+/K+AT- 4. As a result Na+/K+ ATPase activity falls.
Pase moves sodium across the basolateral membrane of the 5. Up-regulation of proton ATPase returns fish t o the freshwater
gills of freshwater fish. condition.
The gill epithelium of freshwater fish also possess chlo-
ride cells, which mediate uptake of Ca2+from the water.
These cells, which differ from the chloride cells in marine
teleost fish, have an anion transporter on the apical mem- uptake is no longer required. The influx of sodium from
brane and high levels of proton ATPase within the cell. seawater causes plasma Na+ levels to rise, which in turn
They may be involved in the uptake of C1-, as well as Ca2+, stimulates secretion of cortisol (Figure 14-43A). Cortisol, -
Figure 14-43 Cortisol plays a major role in inducing the physiological cluding an increase in the gill Na+/K+ATPase activity.As this activity in-
adaptation that occurs when coho salmon are transferred from freshwa- creases, sodium secretion from the gill rises; thus, after several days in
ter to seawater. (A) Initially after a f~shmoves into seawater, the plasma seawater, plasma sodium returns to values close to those observed in
sodium level begins to rise, stimulating secretion of cortisol. (B)The spike freshwater fish. [Unpublished data supplied by N. M. Whiteley and
in plasma cortisol levels mediates a number of changes in the gills in- J. M. Wilson.]
Secretory-Reabsorption Systems
Insects can survive in both freshwater and arid terrestrial
environments; given their often large surface-to-volume ra-
Crayfish antennal "gland"
tios, the osmotic demands placed on these insects can be ex-
Figure 14-44 Osmoregulation in some invertebrates depends on filtra-
treme. The locust, for example, has a large capacity to reg-
tion-reabsorption organs that differ structurally from the mammalian kid- ulate the ionic strength of the hemolymph (blood). During
ney but are functionally analogous. In this schematic diagram of the cray- dehydration the hemolymph volume may decrease by up to
fish antennal gland, the osmotically active tissues are shown in color. 90%, but its ionic composition is maintained. In addition,
Filtration of the blood produces the initial excretory fluid, which then is when this insect is given solutions to drink that range in os-
modified by selective reabsorption of various substances. [Adapted from
Phillips, 1975.1
motic strength from that of seawater to that of tapwater,
hemolymph osmotic pressure changes by only 30%. This
capacity to regulate hemolymph composition depends on a
in regulating the concentration of ions (e.g., Mg2+)in the secretory-type osmoregulatory system.
hernolymph. In broad outline, the osmoregulatory system of locusts
Since the final urine in mollusks and crustaceans differs and other insects consists of Malpighian tubules and the
in composition from the initial filtrate, there must be either hindgut (ileum, colon, and rectum). The closed ends of
secretion of substances into the filtrate or reabsorption of the long, thin Malpighian tubules lie in the hemocoel (the
substances from the filtrate. The reabsorption of elec- blood-containing body cavity); the tubules empty into the
trolytes is well established in freshwater species, for the fi- alimentary canal at the junction of the midgut and hindgut
nal urine has a lower salt concentration than either the (Figure 14-45).The secretion formed in the tubules passes
plasma or the filtrate. Glucose must also be reabsorbed, into the hindgut, where it is dehydrated and passes into the
since it is present in the plasma and in the filtrate but is ei- rectum and voided as a concentrated urine through the
ther absent or present at very low concentrations in the fi- anus. The presence of a tracheal system for respiration in
nal urine. insects (describedin Chapter 13)diminishes the importance
It is interesting that the filtration-reabsorption type of of an efficient circulatory system. As a consequence, the
osmoregulatory system has appeared in at least three phyla Malpighian tubules do not receive a direct, pressurized ar-
(Mollusca,Arthropods, Chordata) and perhaps more. This terial blood supply, as the mammalian nephron does. In-
kind of system has the important advantage that all the stead, they are surrounded with blood, which is at a pres-
low-molecular-weight constituents of the plasma are fil- sure essentially no greater than the pressure within the
tered into the ultrafiltrate in proportion to their concentra- tubules. Since there is no significant pressure differential
tion in the plasma. Such physiologically important mole- across the walls of the Malpighian tubules, filtration can-
cules as glucose and, in freshwater animals, such ions as not play a role in urine formation in insects. Instead, the
Na+, K+, C1-, and Ca2+are subsequently removed from urine must be formed entirely by secretion, with the subse-
the filtrate by tubular reabsorption, leaving toxic sub- quent reabsorption of some constituents of the secreted
stances or unimportant molecules behind to be excreted in fluid. This process is analogous to the formation of urine by
the urine. This process avoids the need for active transport secretion in the aglomerular kidneys of marine teleosts. The
into the urine of toxic metabolites or, for that matter, un- serosal surface of the Malpighian tubule exhibits a profu-
natural, man-made substances of a neutral or toxic nature sion of microvilli and mitochondria, a specialization often
encountered in the environment. Thus, an advantage of the associated with a highly active secretory epithelium.
filtration-reabsorption system is that it permits the excre-
tion of unknown and unwanted chemicals taken in from
the environment without the necessity for a large number
of distinct transport systems.
A disadvantage of the filtration-reabsorption osmoreg-
ulatory system is its high energetic cost for the organism.
Midgut 2 colon 7 Rectum
~ a l p i ~ h i atubule
n
Figure 14-45 Osmoregulat~onIn Insects ~nvolvesa secretory-reab- trated by the reabsorpt~onof water, although Ions also are reabsorbed,
sorpt~onmechan~sm(A) External s ~ d evlew and cross-sect~onalvlew the excreted urine 1s hyperton~cto the hemolymph The arrows 1nd1-
of locust (B) S ~ m p l ~ f ~
d~agram
ed showlng relat~onof Malp~gh~antubule cate the clrcular pathway of water and Ion movement The Insect body
to gut of the locust The pre-urlne 1s produced by secretion Into the contalns numerous Malpcgh~antubules, even though only two are
lumen of the Malp~gh~an tubules, wh~chIle In the blood-conta~n~ng dep~cted
hemocoel The pre-urlne flows ~ n t othe rectum, where ~tIS concen-
The details of urine formation by tubular secretion dif- mands. The fluid formed in the Malpighian tubules passes
fer among different insects, but some major features seem into the hindgut, where several important changes in its
to be common throughout. KC1 and, to a lesser extent, composition occur. In the hindgut, water and ions are re-
NaCl are transported from the hemocoel into the tubular moved in amounts that maintain the proper composition
lumen, along with such waste products of nitrogen metab- of the hernolymph. Thus, it is in the hindgut that the com-
olism as uric acid and allantoin. It appears that the trans- position of the final urine is determined. The water and
port of K+ is the major driving force for the formation of ions removed from the urine by the hindgut are transferred
the pre-urine in the Malpighian tubules, with most of the through intimate connections to the lumen of the
other substances following passively. This has been con- Malpighian tubules. These substances are thereby retained
cluded from the following observations: and recycled in the Malpighian tubule-hindgut circuit
(see Figure 14-45B).
The pre-urine is isotonic or slightly hypertonic relative The most complete study of the osmoregulatory be-
to the hemolymph. havior of the hindgut has been done with the desert locust
Schistocerca. The serosal surface of the ileum and rectum
The pre-urine has a high K+ concentration in all insects. is a highly specialized secretory epithelium (Figure 14-46).
The rate of pre-urine formation is a function of K+ When a solution similar to hernolymph is injected into the
concentration in the fluid surrounding the tubule, hindgut of this insect, water, Kf, Na+, and C1- are ab-
higher K+ concentrations producing more rapid pre- sorbed into the surrounding hemolymph. Evidence from
urine accumulation. electrical measurements suggests that the ions are trans-
ported actively with water following. An electrogenic
The formation of pre-urine is largely independent of the chloride pump and potassium channels in the apical mem-
Na+ concentration of the surrounding fluid. brane appear to mediate KC1 uptake from the hindgut lu-
men into the cells of the gut lining. Sodium uptake from
Although K+ is osmotically the most important substance the lumen is coupled to amino acid uptake andlor ammo-
actively transported, there is evidence that active transport nium ion excretion. The KC1 then moves from the cell into
plays an important role in the secretion of uric acid and the hemolymph via appropriate channels in the basolat-
other nitrogenous wastes. era1 membrane, while sodium is removed from the cell
The pre-urine formed in the Malpighian tubules is into the hemolymph by a Na+/K+ATPase (Figure 14-47).
relatively uniform in composition from one species to Acid is excreted into the lumen of the hindgut by a proton
another, and in each species it remains isotonic relative ATPase. The locust hindgut is capable of removing a large
to the hemolymph under different osmoregulatory de- amount of ions plus water, leaving behind an excess of
I O N I C A N D OSMOTIC BALANCE 619
...............................................................................
Lumen of Figure 14-46 The h~ndgutof Insects 1s spe-
c~al~zedfor transport of water and lons from the
lumen to the surround~nghemocoel Shown
here are the ultrastructural organlzatlon and
Ap~calmembrane gross d~mens~ons of the ep~thel~um of the
Ileum and rectum In the locust, both of wh~ch
are lnvolved In reabsorpt~onNote the exten-
sive ~nfold~ng of the ap~calmembrane and ex-
tensive lateral ~ntercellularspaces [Adapted
mbrane Infolds from lrvlne et al , 1988 ]
Lateral scalarlform
Dllated lntercellular
layer ~ntercellular
spaces Intercellular slnus
(hemocoel)
Subepltheltal space
(hemocoel)
Basal cell
ions and waste products so that the excreted urine is osmotically from the rectum into the Malpighian tubules
hypertonic with an osmolarity up to four times that of because of the KC1 gradient produced by active transport.
the blood. The direction of bulk flow in these compartments is such
In the mealworm Tenebrio, the urine-to-blood osmo- that the osmotic gradient along the length of the complex
larity ratio can be as high as 10, which is comparable to the is maximized, with the absolute osmolarities highest to-
concentrating ability of the most efficient mammalian kid- ward the anal end of the rectum. This gradient may allow
neys. It has been suggested that uphill transport of water in the concentrations near the anal end to exceed those of the
Tenebrio and some other species results from a counter- hemolymph by several times.
current arrangement of the Malpighian tubules, the peri- There is evidence for the feedback regulation of
rectal space, and the rectum (Figure 14-48).Water is drawn osmolarity among the invertebrates, especially in insects.
The bug Rhodnius becomes bloated after sucking blood
from a mammalian host. Within 2-3 minutes, the
Rectal lumen Hemocoel Malpighian tubules increase their secretion of fluid by
more than a thousand times, producing a copious urine.
Artificially bloating the insect with a saline solution does
not produce such diuresis in an unfed Rhodnius. It has
also been found that isolated Malpighian tubules im-
mersed in the hemolymph of unfed individuals remain
quiescent, but if immersed in the hemolymph of a recently
fed Rhodnius they produce a copious secretion. A factor
that stimulates the secretion of these tubules can be
extracted from the neural tissue containing the cell bodies
or axons of neurosecretory cells, primarily those of the
metathoracic ganglion. Thus, it appears that these cells
release a diuretic hormone in response to a factor present
in the ingested blood. The only identified neurohumor
that stimulates the diuretic action of the neurosecretory
cells is serotonin. Similar findings in other insect species
suggest that diuretic and antidiuretic hormones produced
- -
Perirectal space
Rectal lumen
T
Anus
* L
- Amino acids ,
/ Retention as
osmolyte \ + ma~ntenance
some excretlo
&
Excreted
Figure 14-50 Although exceptions occur, water availability correlates main nitrogen excretory product used by an animal, it can be classified as
with the predominant nitrogen excretory product (pink boxes) found in ammonotelic, ureotelic, oruricotelic. In some animals certain nitrogenous
different animals. This general overview of nitrogen metabolism and ex- compounds act as osmolytes, which are substances used to adjust body
cretion in animals shows the polnts at which they differ. Based on the osmolarity. [Adapted from Wright, 1995.1
uric acid via their kidney. That is, the type of excretion is Ammonia-Excreting (Ammonotelic) Animals
generally related to habitat: Terrestrial birds excrete about Most teleosts and aquatic invertebrates are ammonotelic;
90% of their nitrogenous wastes as uric acid and only that is, they excrete their nitrogenous wastes primarily as
3%-4% as ammonia, but semiaquatic birds, such as ammonia, producing little or no urea. As just noted, most
ducks, excrete only 50% of their nitrogenous wastes as uric land animals convert nitrogenous wastes to either uric acid
acid and 30% as ammonia. Mammals excrete most of their or urea to save water. Interesting exceptions are the terres-
waste as urea. Frog tadpoles are aquatic and excrete am- trial isopod (sowbug),as well as some terrestrial snails and
monia; after they metamorphose into the terrestrial adult crabs; these animals eliminate a significant portion of their
form, they excrete urea. Avian embryos produce ammonia nitrogen waste by ammonia volatilization.
for the first day or so, and then switch to uric acid, which is Cell membranes are generally permeable to un-ionized
deposited within the egg as an insoluble solid and thus has ammonia (NH,) but not very permeable to ammonium
no effect on the osmolarity of the precious little fluid con- (NH,+) ions. Ammonia excretion is due largely to the pas-
tained in the egg. Lizards and snakes have various devel- sive diffusion of un-ionized ammonia. In most teleosts,
opmental schedules for switching from the production of nearly all ammonia is excreted as NH,.The associated ex-
ammonia and urea to the production of mainly uric acid. cretion of H + and carbon dioxide acidifies the water next
In species that lay their eggs in moist sand, the switch to to the gill surface, trapping NH, as the largely impermeant
uric acid production occurs late in development, but before NH,+ and enhancing ammonia excretion. Some mem-
hatching. The switch to uric acid production is a kind of branes, however, have a low permeability to NH3, as well
biochemical metamorphosis that prepares the organism for as NH,+. Membranes of Xenopus eggs and those of the
a dry, terrestrial habitat. It is evident, however, that there cells of the thick ascending limb of the loop of Henle in the
are overlaps of different excretory products in animals in mammalian kidney are examples of structures having a low
similar environments. NH, permeability.
Amino groups of various amino acids are trans- toxic, should act as the amino-group carrier through
ferred, with the aid of a transaminase enzyme, to gluta- blood and tissues until its deamination in the ammonotelic
mate, which is then deaminated to form ammonium ions kidney.
and a-ketoglutarate in the liver. In the liver, glutamate is A blood concentration of only 0.05 mmo1.L-' am-
also converted to glutamine, which is much less toxic than monia is toxic to most mammals, causing convulsions
ammonia and crosses membranes easily but is not nor- and death. Similar acute toxic effects have been observed
mally excreted in any quantity. Although mammals ex- in many other animals, including birds, reptiles, and
crete most nitrogenous waste as urea, they excrete small fish. Mexican guano bats are unusual among mammals
amounts of ammonia in the urine. The less toxic gluta- in that they can withstand very high levels of ammonia
mine, rather than ammonia, is released from the mam- (1800 ppm) in the atmosphere of the caves in which
malian liver into the blood and taken up by the kidney. they live. This level is sufficient to kill humans, so enter
The glutamine is then deaminated in the cells of the kid- guano bat caves with care! The toxicity of NH, is due
ney tubules, and ammonia is liberated into the tubular in part to the elevation of p H it produces, which causes
fluid. The excreted ammonia can take up a proton to form changes in the tertiary structure of proteins. Ammonia also
the NH,+, which cannot diffuse back into the tubular cells interferes with some ion-transport mechanisms, because
and thus leaves the body via the urine (see Figure 14-30). NH,+ substitutes for K+ in some cases. Ammonia can also
-
Since ammonia in both its free and ionized forms is highly effect brain blood flow and some aspects of synaptic trans-
toxic, it makes good sense that glutamine, which is non- mission, particularly glutamate metabolism.
W#-C
T l3 t nvr I
Carbamoyl
,C=O
I
- 1
synthetase Ornithine HN
p h O s p h ~ carbamoyl wansferase 1
;
HC-NH,
\
I
COOH
L-Ornlthine H
HOOC-CH~-C-COOH
iH2
L-Aspartlc acid
Argininosucc~nate
synthetase
I COOH
I
C=N-C-H
AHz Argininosucc~nate I
HC-NH,
Arginase lyase
I I
FH2 COOH
I L-Argininosuccinate
HC-NH,
I
COOH
Figure 14-51 Urea IS formed bythe orn~th~ne-urea cycle In all vertebrates mals consume more energy In the excretion of nltrogen than do other
except teleosts Because ATP IS required for the f~rststep, ureotel~canl- an~mals.
molecule of CO, are added to ornithine to form arginine.
are the effects of variations in t The enzyme arginase, which is present in relatively large
quantities in these animals, then catalyzes removal of the
urea molecule from arginine, regenerating ornithine.
The Lake Magadi tilapia, Oreochromis alcalicus gra-
hami, is a completely aquatic freshwater teleost fish; unlike
Some squid, shrimps, and tunicates sequester NH,+ in most teleosts, it excretes all nitrogenous waste as urea. The
high concentration in specialized acidkied chambers that act high p H of Lake Magadi (about 10) impairs ammonia ex-
as a float making the animal more buoyant. In the floats of cretion to an extent that leads to ammonia accumulation
these marine animals, NH,+ is substituted for heavier Ca2+, and death in other fish. Oreochromis alcalicus grahami can
Mg2+,and S042- ions (see Figure 14-50).Ammonium lev- live in Lake Magadi because it converts ammonia to urea via
els in the floats are very high and the tissues that make up the the ornithine-urea cycle, thereby avoiding ammonia toxicity.
float must be resistant to the toxic actions of ammonia. Am- Elasmobranch fish use urea produced from ammonia via the
monia levels in other regions of the body are relatively low. ornithine-urea cycle to increase their body osmolarity; they
also excrete most of their nitrogenous waste as urea across
Urea-Excreting (Ureotelic) Animals their gills. So not all aquatic animals excrete ammonia.
Ureotelic animals excrete most nitrogen wastes as urea, Most teleosts and many invertebrates utilize the so-
which is quite soluble in water, is far less toxic than ammo- called uricolytic pathway in which urea is produced from
nia, and requires much less water for excretion. Moreover, uric acid that either arises from a transamination via aspar-
urea contains two nitrogen atoms per molecule. Ureotelic tate or is produced during nucleic acid metabolism. In this
animals utilize one of two pathways for urea formation. pathway, uric acid is converted first to allantoin and allan-
With the exception of most teleost fishes, vertebrates syn- toic acid with the help of the enzymes uricase and allan-
thesize urea primarily in the liver via the ornithine-urea toinase, respectively, and then to urea using allantoicase
cycle (Figure 14-51). In this cycle, two -NH, groups and a (Figure 14-52). During evolution most mammals have lost
'
duced vta the ur~colyttcpathway. U r ~ ca c ~ d
arlses from a purlne rlng that IS synthesized
by a complex unlon of aspartlc ac~d,form~c
I Transaminase
(Purine ring)
Uric acid
Uricase
+ 0 2
N"2
I
O=C COOH C=O
I I I
< "20 HN-C- NH
Allanto~case H Allantolnase H H
Allantoic a c ~ d Allanto~n
the ability to produce allantoicase and allantoinase; homi- primeval seas that are believed to have been the setting for
noid primates also cannot synthesize uricase and thus ex- the early evolution of animal life. The ability of many ani-
crete uric acid as an end product of nucleic acid metabolism. mals to regulate the composition of their internal environ-
Uric acid excretion is normally only about 1% by weight of ment is closely correlated with their ability to occupy eco-
urea excretion in humans. Should uric acid production or logical environments that are osmotically at odds with the
intake increase, however, uric acid levels in the blood may osmotic requirements of their tissues. Osmoregulation re-
rise because excretion may be compromised due to low uric quires the exchange of salts and water between the extra-
acid solubility, especially if urine volume is small. The low cellular environment and the external environment to com-
solubility may also result in the precipitation of uric acid pensate for obligatory, or uncontrolled, losses and gains.
crystals when levels of uric acid rise in the blood, which in The transport of solutes and water across epithelial layers
turn causes the painful condition called gout. is fundamental to all osmoregulatory activity. The obliga-
Urea crosses membranes either through aqueous pores tory exchange of water depends on (1)the osmotic gradi-
or via specialized membrane transporters, as lipid mem- ent that exists between the internal and external environ-
branes are not very permeable to urea. Specific urea trans- ments, (2)the surface-to-volume ratio of the animal, (3)the
porters are thought to be present in elasmobranchs and permeability of the integument, (4)the intake of food and
several other vertebrates, including mammals. Urea trans- water, ( 5 )the evaporative losses required for thermoregu-
porters are widely distributed and in some instances may be lation, and (6) the disposal of digestive and metabolic
involved in rapid urea transport to stabilize cell volume in wastes in urine and feces.
the face of an osmotic shock. Marine and terrestrial animals are faced with dehydra-
tion, whereas freshwater animals must prevent hydration
Uric Acid-Excreting (Uricotelic) Animals by uncontrolled osmotic uptake of water. Marine birds,
Uricotelic animals-birds, reptiles, and most terrestrial reptiles, and teleosts replace lost water by drinking seawa-
arthropods-excrete nitrogen chiefly in the form of uric ter and actively secreting salt through secretory epithelia.
acid or the closely related compound guanine. Uric acid Freshwater fishes do not drink water; they replace lost salts
and guanine have the advantage of carrying away four ni- by active uptake. Birds and mammals are the only verte-
trogen atoms per molecule. The nitrogen atoms incor- brates that secrete a hypertonic urine. Many desert species
porated into uric acid ultimately arise from the breakdown in addition utilize mechanisms for minimizing respiratory
of the amino acids glycine, aspartate, and glutamine (see water loss.
Figure 14-52).Because these animals lack uricase, they can- Most vertebrate kidneys utilize filtration, reabsorption,
not break down uric acid. Thus the catalysis of nitrogenous and secretion to form urine. A countercurrent mechanism
molecules is terminated at uric acid, which largely precipi- present in mammalian and avian kidneys allows produc-
tates because of its low solubility and is excreted as the end tion of a hypertonic urine. The filtration of the plasma in
product, requiring very little urinary water. In general, uri- the glomerulus is dependent on arterial pressure. Crystal-
cotelic animals are adapted to conditions of limited avail- loids and small organic molecules are filtered, leaving blood
ability of water. cells and large molecules behind. Salts and organic mole-
Uric acid is transported from the blood into the cells of cules such as sugars are partially reabsorbed from the
the renal tubule via a urate-anion exchanger or via a urate glomerular filtrate in the renal tubules, and certain sub-
uniporter. It then moves from the cells into the lumen of the stances are secreted into the tubular fluid. A countercurrent
tubule down an electrochemical gradient and is excreted in multiplier system that includes the collecting duct and the
the urine. Urate transport competes with para-aminohip- loop of Henle sets up a steep extracellular concentration
purate transport in the kidney tubule of birds but not in gradient of salt and urea that extends deep into the medulla
reptiles. of the mammalian kidney. Water is drawn osmotically out
Two unusual amphibians are the arid-land toads Chi- of the collecting duct as it passes through high medullary
romantis xerampelina and Phyllomedusa sauvagii. These concentrations of salt and urea toward the renal pelvis.
toads not only have an extremely low evaporative water Endocrine control of the water permeability of the collect-
loss from their skin but, like reptiles, excrete nitrogen as ing duct determines the volume of water reabsorbed and
uric acid rather than ammonia or urea as most other am- retained in the circulation. The final urine, then, is the
phibians do. The low solubility of uric acid causes it to pre- product of filtration, reabsorption, and secretion. These
cipitate readily in the cloaca, and allows these toads, like processes allow the urinary composition to depart strongly
reptiles and birds, to minimize the volume of urine neces- from the proportions of substances occurring in the blood.
sary to eliminate their excess nitrogen. The formation of urine follows the same major outline
in all or most vertebrates and invertebrates. A pre-urine is
formed that contains essentially all the small molecules and
SUMMARY
ions found in the blood. In most vertebrates and in the crus-
The extracellular environment in many marine and non- taceans and mollusks, this formation is accomplished by
marine animals broadly resembles dilute seawater. This ultrafiltration; in insects, by the secretion through the ep-
similarity may have had its origin in the shallow, dilute ithelium of the Malpighian tubules of KCl, NaCI, and
IONIC A N D OSMOTIC BALANCE 625
......................................
phosphate, with water and other small molecules, such as ing the loop, in what way is the final urine made hy-
amino acids and sugars, following passively by osmosis and pertonic in the mammalian kidney?
diffusion down their concentration gradients. The pre- Explain why the consumption of 1 liter of beer will
urine is subsequently modified by the selective reabsorption lead to a greater urine production than an equal vol-
of ions and water and, in some animals, by secretion of ume of water.
waste substances into the lumen of the nephron by the What role does the kidney play in the regulation of
tubular epithelium. blood pressure?
Birds and reptiles can drink seawater, excreting the salt Discuss the role of the kidney in the control of
load through a nasal salt gland. Elasmobranchs excrete salt plasma pH.
via a rectal gland, which is made up of salt-secreting cells How do insects produce concentrated, hypertonic
similar to those found in the thick ascending limb of the urine and excrement?
loop of Henle in the mammalian kidney, the avian and rep- In the course of evolution, terrestrial organisms have
tilian salt gland, and the chloride cell in the gills of marine come to excrete mainly uric acid and urea rather than
teleosts. The hormonal regulation of the activity of these ammonia. What are the adaptive reasons for such a
cells is also similar in sharks, birds, reptiles, and mammals. change?
The gills of teleost fishes and many invertebrates perform Explain why gulls can drink seawater and survive but
osmoregulation by active transport of salts, the direction of humans cannot.
transport being inward in freshwater fish and outward in After the injection of inulin into a small mammal,
marine fish. the plasma inulin concentration was found to
The nitrogen produced in the catabolism of amino be 1 rng.mlk1, the concentration in the urine
acids and proteins is concentrated into one of three forms 10 mg-ml-l, and the urine flow rate through the
of nitrogenous waste, depending on the osmotic environ- ureter 10 ml. h-'. What was the rate of plasma fil-
ment of different animal groups. Ammonia, highly toxic tration and the clearance in milliliters per minute?
and soluble, requires large volumes of water for dilution How much water was reabsorbed in the tubules per
and subsequent excretion across the gills of teleosts. Uric hour?
acid is less toxic and poorly soluble; it is excreted as a What evidence is there that the mammalian nephron
semisolid suspension via the kidney of birds and reptiles. employs tubular secretion as one means of eliminat-
Urea is the least toxic and its excretion requires a moder- ing substances into the urine?
ately small amount of water. Mammals largely convert why is a countercurrent system more efficient in
their nitrogenous wastes into urea, which is excreted in the physical transport and transfer than a system in which
urine; elasmobranchs use urea as an osmotic agent in their fluids in opposed vessels flow in the same direction?
blood and excrete most excess nitrogen as urea across What are the similarities and differences between the
their gills. elasmobranch rectal gland and the bird salt gland?
c-
- - #-- ---- Photosynthetic
&- -- autotroph
CO, + H,O
4c---%&
, 3-
% *_ -..--
Heterotroph
Figure 15-1 Two trophic levels occur in this generalizedflow diagram of in metabolism. Chemical energy content is at its peak following the pho-
chemical energy through a food chain. The flowchart begins with the tosynthetic production of sugars. When the plant material is consumed
photosynthetic formation of high-energy-content molecules (sugars) by a heterotroph, some chemical energy IS converted to heat and thus
from low-energy-content raw materials (CO, and H,O) in plants. Oxida- is lost as a direct source of energy for driving biological processes. This
tion of carbon compounds yields free energy coupled to the synthesis of food chain has only two trophic levels, but most food chains have many
high-energy compounds, such as ATP, used as common energy currency more intervening levels.
animals follow a more active sequence, which in the ex- Some free-living protozoans and invertebrates derive
treme of many carnivores (meat eaters) includes searching, part of their nutrients by direct surface uptake from the sur-
stalking, pouncing, capturing, and killing. rounding medium. Small molecules such as amino acids are
taken up from dilute solution by transport mechanisms (de-
Food Absorption through Exterior Body Surfaces scribed in Chapter 4), against what can often be a huge
The feeding method that is least dependent on specialized concentration gradient. In some of these organisms, larger
capture and digestive organs involves absorption of nutri- molecules or particles are taken up by a bulk process such
ents directly across the body wall. Certain protozoans, en- as phagocytosis, which is described next.
doparasites (animals that live within other animals), and
aquatic invertebrates are able to take up nutrient molecules Endocytosis
from the surrounding medium directly through their soft Endocytosis represents a more active form of "feeding"
body wall. Endoparasites such as parasitic protozoans, than passive absorption directly across the body wall. Like
tapeworms, flukes, and certain mollusks and crustaceans direct nutrient absorption, however, it occurs at the local
are surrounded by host tissues or by alimentary canal flu- cellular rather than tissue or organismal level. Endocytosis
ids, both of which are high in nutrients. Tapeworms, which includes two processes, phagocytosis ("cell eating") and
may be many meters long, lack even a rudimentary diges- pinocytosis ("cell drinking"). In phagocytosis, pseudopod-
tive system. Tapeworms evolved from a primitive flatworm like protuberances extend out and envelope relatively large
that lacked a body cavity (i.e., was acoelomic). However, nutrient particles. Pinocytosis occurs when a smaller par-
some endoparasites appear to have secondarily lost the di- ticle binds to the cell surface and the plasma membrane in-
gestive apparatus present in their ancestors. For example, vaginates (folds inward) under it, forming an endocytotic
parasitic crustaceans, which belong to the cirripeds (bar- cavity. Whether captured by phagocytosis or pinocytosis,
nacle group), lack an alimentary canal, but they appear to the morsel is then engulfed in a membrane-enclosed vesicle
have evolved from nonparasitic ancestors possessing a gut. that pinches off from the bottom of the cavity.
ACQUIRING ENERGY: FEEDING, DIGESTION, A N D METABOLISM 629
...............................................................................
The vesicle (or food vacuole in Protozoa) fuses with Oscu~um
lysosomes, organelles containing intracellular digestive en-
zymes, whereafter it is called a secondary vacuole. After
digestion, the contents of the vacuole pass through the vac-
uole wall into the cytoplasm. The remaining undigested
material is excreted externally by exocytosis, essentially
a reverse process of pinocytosis. Feeding by pinocytosis
and phagocytosis is familiar in protozoans such as Para- ath of water
mecium, but also occurs in the lining of the alimentary drawn into
flagellated
canals and other tissues of many multicellular animals. chambers
Filter Feeding
Many aquatic animals use filter feeding, also called sus-
pension feedmg, to capture food. Food items (usuallyphy-
toplankton or zooplankton) are carried to specialized en-
trapment devices either on the body surface or within it. Figure 15-2 Water flows in an organized fashion through syconoid
Most marine filter feeders are small, sessile animals, sponges. In this diagrammatic section, red arrows indicate water flow. A
such as sponges, brachiopods, lamellibranchs, and tuni- significant proportion of this flow results from the reduction in hydrosta-
cates. Food items are carried along on water currents that tic pressure at the osculum due to the Bernoulli effect, produced by the
either occur naturally or are generated by the movements transverse water currents (black arrows) flowing over the osculum at ele-
vated velocity.Water flow is also generated from the activity of flagellated
of body parts of the filter-feeding animal itself, such as cilia
choanocytes that line the flagellated chambers (and give them their
or flagella. Brachiopods respond behaviorally to currents, name).The choanocytes are found in the regions of the flagellated cham-
rotating on their pedal stalks to present the most efficient bers lined in red. Water enters the sponge through the ostia, passes
hydrodynamic orientation for capturing the water current. through the flagellated chambers, and ends up in the inner cavity, the
A number of other sessile animals located in moving wa- spongocoel. Nutrients are then taken up into individual cells by endocy-
tosis. [Adapted from Hyman, 1940; Vogel, 1978.1
ter make use of Bernoulli's effect (i.e., a drop in fluid pres-
sure as fluid velocity increases) to increase the rate of water
flowing through the entrapment sites, at no energy cost to along the filaments (i.e., 90" to the water flow) to the tip of
themselves. An example of such passively assisted filter the gill, where it travels in a special groove under ciliary
feeding is seen in sponges (Figure 15-2). The flow of water power toward the mouth in a rope-like string of mucus.
across the large terminal opening causes a drop in pressure Sand and other indigestible particles are sorted out and re-
(Bernoulli's effect) outside the osculum. As a result, water jected (presumably on the basis of texture), passing out
is drawn out of the sponge through the osculum, and is with the water leaving the exhalant siphon.
drawn in through the numerous ostia (mouth-like open- Non-sessile animals filter-feed by various mechanisms.
ings) in the body wall. The drop in pressure is facilitated by A number of fishes are planktivorous, using modified gill
the shape of the sponge's exterior, which causes the water
over the osculum to flow with greater velocity than the wa-
ter flowing past the ostia. Food particles, swept into the os- Ctenidium Adduct,or muscle
Labrum
Mandible
Maxilla
Hypophatynx
Salivary
channel
Labium
Figure 15-5 Sucking insects use tubular mouthparts for feeding. (A) lat-
eral view of the head of a mosquito, with the mouthparts separated for
identification. (6) Cross-sectionof the assembled mouthparts of the mos- Figure 15-6 The anterior limbs of arthropods are often modified for seiz-
quito, showing separate channels for blood to move superiorly to the ing prey and holding it while the mouthparts tear off small pieces to be
mouth, and for saliva to move inferiorly to the wound. (C) Lateral view of swallowed. (A) A mantispid insect (order Neuroptera).(B) Lobsters (order
the head of a moth, showing the sucking mouthparts curled up between Decapoda) have one claw modified for tearing and one modified for
feeding bouts. [Adapted from Rupert and Barnes, 1994.1 crushing.
Generalized
placental
mammal
n African
Figure 15-7 Rattlesnakes have modified teeth, known as fangs, which
they use to inject venom into their prey. These side views of a rattlesnake
skull show (A) a non-striking position, with the jaws only partially open
and the hinged fangs folded into the roof of the mouth, and (B) a striking
position, in which the jaws are open wide and the fangs extended. The
extraordinaryflexibility of the lower jaws allows the snake to swallow prey
whole after injecting it with deadly venom. [Adapted from Parker, 1963.1
Herbivores Carnivores
Omnivores
Filter-feeding
(goose)
Seed-eating /
(crossbill)
(heron)
( Flesh-eatlng
(hawk)
Figure 15-9 Bird beaks are adapted to suit herbivory, ornnlvoty, and carnivoty. [Adapted from Marshall and Hughes, 1980.1
634 INTEGRATION OF PHYSIOLOGICAL SYSTEMS
......................................
neurotransmitter at the motor endplate in muscle. A neu-
rotoxin, a-bungarotoxin (see Spotlight 6-3), found in the
venom of the cobra-like krait, binds to nicotinic acetyl-
choline (ACh)receptors, thereby blocking neuromuscular
transmission in vertebrates. The venoms of various species
of rattlesnake contain hemolytic (blood cell-destroying)
substances.
Toxins, although highly effective, are generally expen-
Neyatocysts
\\k ~y stalk
sive to produce. Usually carefully measured doses of toxins
are delivered during a bite or sting. Toxins also must
be specially stored before administration to avoid self-
poisoning. Toxins are generally proteins and, as such, are
rendered harmless by the proteolytic enzymes of the preda-
tor's digestive system when it ingests its poisoned prey.
Figure 15-11 The scorpion Androctonus captures prey and then injects
poison to subdue it. As the prey item is grabbed by the chela, the tail is Figure 15-12 The head of a gastropod mollusk contains a powerful
arched over the scorpion's head to bring the sting into position. The prey radula. (A) The rasp-like radula, revealed in sagittal section, is used for
is then impaled by the sting, which injects the rapidly acting toxin. grazing on vegetation. (B) Protraction of the radula. (C)Retractionof the
[Adapted from Jennings, 1972.1 radula. [Adapted from Rupert and Barnes, 1994.1
ified for grinding plant material. Plants (especially some (Figure 15-13, left). Coelenterates, for example, have a
grasses)contain relatively large amounts of silicates, and can blind tube or cavity, the coelenteron, which opens only at a
be tremendously abrasive. Consequently, the molars of her- "mouth" that serves also for the expulsion of undigested
bivores often are coated in especially tough enamel to resist remains. In all phyla higher than the flatworms, ingested
wear. Alternatively, some herbivores such as small rodents material passes through a hollow, tubular cavity-the
(microtines)have continuously growing, rootless teeth. alimentary canal-extending through the organism and
open at both ends. Processing goes on continuously,
rather than in pulses, with new food being ingested while
OVERVIEW OF ALIMENTARY SYSTEMS older food is still being processed. Some alimentary canals
Alimentary systems play an essential role in providing nour- can be modeled as ideal continuous-flow, stirred-tank
ishment through digesting and absorbing food, and remov- reactors, in which food is continually added and mixed into
ing from the body indigestible materials and toxic by-prod- a homogeneous mass, and the products of digestion
ucts of digestion. The most primitive "alimentary system" is are continuously eliminated, overflowing from the reactor
the plasma membrane of unicellular organisms, in which mi- (Figure 15-13, middle). An example of such a reactor is the
croscopic food particles are engulfed, undigested, by endo- forestomach of ruminants. The third way of processing
cytosis directly into the cell itself. Once in the cell, food par- food is in a plug-flow reactor, in which a bolus (a discrete
ticles undergo intracellular digestion by acids and enzymes. plug or collection) of food is progressively digested as it
More complex multicellular animals rely primarily on ex- winds its way through a long, tube-like digestive reactor
tracellular digestion carried out by true alimentary systems. (Figure 15-13, right). Unlike the stirred-tank reactor, its
From an anatomical perspective, there are myriad composition varies according to its position along the re-
designs of alimentary systems. However, from a physiolog- actor tube. The small intestine of many vertebrates func-
ical perspective, alimentary systems fall into one of three tions as a plug-flow reactor. It is important to recognize that
categories on the basis of how they process food in a "re- many animals combine features of both continuous- and
actor," or site of chemical digestion. So-called batch reac- plug-flow reactors. As you will see below, in many animals
tors are blind tubes or cavities that receive food and elimi- chemical digestion begins in the stomach, configured as a
nate wastes in a pulsed fashion; that is, one batch is continuous-flow, stirred-tank reactor, and then continues
processed and eliminated before the next one is brought in on into the small intestine, configured as a plug-flow reactor.
Third compartment
Gastrovascular
cavity
Figure 15-13 Digestive systems are functionally classified according to tine of many vertebrates acts as a plug-flow reactor, which may function
the type of chemical reactor they form. (Left) Batch reactors are found in in addition to the stomach. [From Hume, 1989; adapted from Penry and
simple organisms like Hydra. (Middle) Ruminants have a continuous-flow, Jurnars, 1987.1
stirred-tank reactor in the form of a forestomach. (RightjThe small intes-
636 INTEGRATION OF PHYSIOLOGICAL SYSTEMS
...............................................................................
A High-quality food B Low-quality food Figure 15-14 The qual~tyof food will greatly
Max~mumrate Influence the tlme requ~redfor d~gest~on
In a
of digest~on Maximum rate cont~nuous-flowd~gest~ve reactor (A) A hlgh-
qual~tyfood requlres m ~ n ~ menergy
al to cap-
of digest~on
ture and eat and, once eaten, 1s qulckly dl-
gested to release large amounts of energy
The maximum rate of d~gest~on occurs at the
polnt of the curved l~new ~ t hthe steepest
slope (B) A low-qual~tyfood requlres consld-
erable energy to capture and eat, and takes a
) long per~odof d~gest~on to yleld only low
amounts of energy [Adapted from Hume,
1989, Sibly, 1981 ]
Mean retention t~me Mean retention time
of food of food
It is critically important that the design of the alimen- (4)absorbing water and defecating. Representative alimen-
tary canal and the reactors it contains matches well with the tary canals from the different invertebrate and vertebrate
quality of the food that the animal routinely eats. A high- classes are illustrated in Figures 15-16 and 15-17,respectively.
quality food can release maximum amounts of energy -.
with
minimal time spent in the digestive reactor, whatever its Ingestion
type (Figure 15-14). A lower-quality food, on the other
hand, requires a longer period of digestion to release its en- Headgut
ergy. This in turn requires longer periods spent in the reac-
tor and longer transit times through the alimentary canal.
As also indicated in Fig. 15-14, the amount of energy spent \ Conducting
in capturing a particular food must also be factored into Foregut
consideration of food quality.
A generalized alimentary canal, or digestive tract, is il-
lustrated in (Figure 15-15). The lumen of this alimentary
canal is topologically external to the body. Sphincters and
other devices guard the entrance to and exit from the canal,
preventing uncontrolled exchange between the lumen and
the external environment. Ingested material is subjected to Midgut
various mechanical, chemical, and bacterial treatments as it
passes through this canal, and digestive juices (primarilyen-
zymes and acids)are mixed with the ingested material at ap- (Acidic secretions)
propriate regions in the alimentary canal. As the ingested
material is first mechanically broken down and then chemi-
cally digested, nutrients undergo absorption and are then
transported into the circulatory system. Undigested, non-ab-
sorbed material is stored briefly until it, along with bacter-
ial remains, is expelled as feces by the process of defecation.
The overall tubular organization of the alimentary
Absorption-+Assimilation
canal is efficient because it allows ingested material to travel (Basic secretions)
in one direction, passing through different regions that can
then be specialized for particular digestive tasks. For ex-
ample, the alimentary canal near the point of ingestion is
often specialized for acid secretion, while more distant re-
gions are alkaline. This regional specialization allows both Hindgut
acid and base secretion to occur at the same time and to Storage of waste .,
Tentacle
Enzymatic Mesoglea
gland cells /
Gastro
cavity
Food vacuole
Branch6
enteron
Gastric
Figure 15-16 Digestive systems of invertebrates show great variation, (C) Digestive system of a prosobranch gastropod mollusk. Arrows show
ranging from simple to highly complex. (A) Section through body wall of ciliary currents and the rotation of the mucous mass. (D) Digestivesystem
Hydra, a coelenterate. The epithelial lining of the coelenteron includes of the cockroach Periplaneta. The proventriculus (or gizzard) contains
phagocytosing cells (called nutritive muscle cells) and gland cells that se- chitinous teeth for grinding food. [Part C from Rupert and Barnes, 1994;
Crete digestive enzymes. (B) Digestive system of a polyclad flatworm. part D from Imms, 1949.1
638 INTEGRATION OF PHYSIOLOGICAL SYSTEMS
...............................................................................
Cyclostome Elasmobranchs Teleost fishes Amphibians
(hagfish) (shark) (bass) (frog)
Figure 15-17 The tube-l~kedigestive system of vertebrates has a basic G, gallbladder; L, liver; LI, large intestine; P, pancreas; PA, pyloric appen-
organizational plan, with common elements of esophagus, stomach, in- dices; SG, spiral gut; SI, small intestine; St, stomach. [From Florey, 1966;
testine, and colon. B, bladder; C, cecum; Cr, crop (gizzard); E, esophagus; adapted from Stempell, 1926.1
A C Q U I R I N G ENERGY: FEEDING, D I G E S T I O N , A N D M E T A B O L I S M 639
...............................................................................
Headgut: Food Reception feedings. In some animals crops are also used to ferment or
The headgut is the anterior (cranial) region of the alimen- digest foods for purposes other than their immediate di-
tary canal, providing an external opening for food entry gestion. Parent birds prepare food in this way to be regur-
(see Figure 15-15).It consists of organs and structures for gitated for their nestlings.
feeding and swallowing, including the mouthparts, buccal
cavity, pharynx, and associated structures such as bills, Stomach
teeth, tongue, and salivary glands. Where a common path- In vertebrates and some invertebrates digestion takes place
way exists leading to both the alimentary canal and the pas- primarily in the stomach and the midgut. The stomach
sageway (e.g., the trachea) ending in the organ of internal serves as a storage site for food, and in many species begins
gas exchange, there may be additional sphincter- or valve- the initial stages of digestion. In most vertebrates, for ex-
like structures that control and divert the flow of ingested ample, the stomach initiates protein digestion by secreting
material and inspired water or air into their respective the enzyme pepsinogen (later converted to pepsin) and hy-
channels. drochloric acid, which provides the highly acidic environ-
Other than in small-particle feeders such as coelenter- ment required for pepsin activation. Contraction of the
ates, flatworms, and sponges, the headgut of most meta- muscular walls of the stomach also provides mechanical
zoans has salivary glands, the secretions of which aid in- mixing of food, saliva, and stomach secretions.
gestion and the mechanical (and often chemical) digestion Stomachs are classified as monogastric or digastric, ac-
-
of food. The primary function of the salivary secretion, cording to the number of chambers they possess. A mono-
saliva, is lubrication to assist swallowing. The lubrication gastric stomach consists of a single strong muscular tube or
is provided in many cases by a slippery mucus of which the sac. Vertebrates that are carnivorous or omnivorous char-
chief constituent is a type of mucopolysaccharide named acteristically have a monogastric stomach (Figure 15-18).
mucin. The saliva often contains additional agents, such as Instead of a stomach, some invertebrates, such as insects
-
digestive enzymes, toxins, and anticoagulants (in blood- (see Figure 15-16D),have outpouchings termed gastric ceca
lapping or blood-sucking animals such as vampire bats and (singular, cecum), which are lined with enzyme-secreting
leeches).(See Chapter 8 for a discussion of salivary glands.) cells, as well as phagocytic cells that engulf partially di-
Tongues, an innovation of the chordates, assist in the gested food and continue the process of digestion. In these
mechanical digestion and swallowing of food. In some an- alimentary systems the processes of digestion and absorp-
imals tongues are used to grasp food. They are also used in tion are completed in the ceca, and the remainder of the al-
chemoreception, bearing gustatory receptors called taste imentary canal is concerned primarily with water and elec-
buds (see Figure 7-16A). Snakes use their forked tongues to trolyte balance and defecation.
take olfactory samples from the air and the substratum, re- Some birds have a tough, muscular gizzard, or crop, or
tracting the tongue to wipe the samples in Jacobson's or- both (see Figure 15-17). Sand, pebbles, or stones are swal-
gan, which consists of a pair of richly innervated chemosen- lowed and then lodge in the gizzard, where they aid in the
sory pits located in the roof of the buccal cavity. Jacobson's grinding of seeds and grains. The proventriculus of insects
organs are found in other reptiles and some amphibians. and the stomach of decapod crustaceans, comparable to
the bird's gizzard, contain grinding apparatuses for chew-
Foregut: Food Conduction, ing swallowed food. Some fish such as mullets also have
Storage, and Digestion gizzards. On the other hand, some fish and larval toads lack
In most species the foregut consists of an esophagus, a tube stomachs altogether, with material from the esophagus en-
that leads from the oral region to the digestive region of the tering into what is functionally the midgut.
alimentary canal, and a stomach (see Figure 15-15). Multichambered digastric stomachs (Figure 15-19) are
found in the mammalian suborder Ruminantia (deer, elk,
Esophagus giraffe, bison, sheep, cattle, etc.). Somewhat similar digas-
The esophagus conducts food from the headgut to the di- tric stomachs occur outside this suborder, in particular in
gestive areas, usually the stomach (see below). In chordates the suborder Tylopoda (camel, llama, alpaca, vicuaiia). Mi-
and some invertebrates, the esophagus conducts the bolus, croorganisms in the first division of the stomach carry out
or mass of chewed food mixed with saliva, by peristaltic fermentation, the anaerobic conversion of organic com-
movement (see Chapter 11)from the buccal cavity or phar- pounds to simpler compounds, yielding energy as ATP. All
ynx. In some animals, this conducting region contains a of the above-named groups carry out rumination, in which
sac-like expanded section, the crop, which is used to store partially digested food is regurgitated (transported back to
food before digestion. The presence of a crop, generally the mouth) for remastication (additional chewing). This
found in animals that feed infrequently, allows quantities process allows the ruminant (a gazelle on the open savanna,
of food to be stored for digestion at a later time. Leeches, for example) to swallow food hastily while grazing and
for example, feed very infrequently, with weeks or months then to chew it more thoroughly later when at rest in a
between feeding periods. However, they ingest large quan- place of relative safety from predators. After the regurgi-
tities of blood at a "sitting," storing the blood for many tated food is chewed, it is swallowed again. This time it
weeks and digesting it in small amounts between their rare passes into the second division of the digastric stomach and
640 INTEGRATION OF PHYSIOLOGICAL SYSTEMS
.................................................................................
A
Esophagus
\ /~~~$$hincter
Duodenum
, .
pylor~r
Abomasum
B I \
\
act as a fermentation vat that receives grazed vegetation.
Bacteria and protozoans in these chambers thrive on the
vegetation, causing extensive digestive breakdown by fer-
mentation of carbohydrates to butyrate, lactate, acetate,
Gastric
pit
and propionate. These products of fermentation, along
with some peptides, amino acids, and short-chain fatty
acids, are absorbed into the bloodstream from the rumen
Goblet fluid. Symbiotic microorganisms grown in the rumen,
cells
along with undigested particles, are passed into the oma-
Parietal sum (absent in the Tylopoda) and then into the abomasum.
cells Only the latter secretes digestive enzymes and is homolo-
gous to the monogastric stomach of non-ruminants.
Fermentation in the stomach is not limited to ruminat-
Gastric,
ing animals. It is found in other animals in which the pas-
gland
sage of food in the stomach is delayed, allowing the growth
of symbiotic microorganisms in a zone anterior to the di-
Chief
cells
gestive stomach, as in the kangaroo and the crops of galli-
form (chicken-like)birds.
Gastric wall Midgut: Chemical Digestion and Absorption
Figure 15-18 The monogastric stomach is a single chamber lined with a In vertebrates, the rnidnut
- is the site for the chemi-
specialized epithelium. (A) Major parts of the mammalian stomach. cal digestion of proteins, fats, and carbohydrates. once di-
(B) Detail of fundic, or gastric, glands lining a single gastric pit. The in-
ner layer of the stomach is lined with thousands of gastric pits, into which
gested to their component molecules, these materials are
the gastric glands open and dump their digestive juices. The epithelium then in the midgut and away
of the gastric gland contains chief (pe~sinogen-secreting)and parietal the alimentary canal in the blood. AS food is ready to pass
(HCI-secreting) cells as well as goblet (mucus-secreting)cells. on from the vertebrate stomach, it is released into the
midgut through the pyloric sphincter, which relaxes as the
peristaltic movements of the stomach squeeze the acidic
begins the second stage of digestion. In this stage hydroly- contents into the duodenum, the initial segment of the
sis takes place with the assistance of digestive enzymes se- small intestine (see Figure 15-18A). Digestion continues in
creted by the stomach lining. the small intestine, generally in an alkaline environment.
The digastric stomach of the Ruminantia (see
Figure 15-19)has four chambers, separated into two divi- General structure and function of the rnidgut
sions. The first division consists of the rumen and reticulum Among the vertebrates, carnivores have shorter and simpler
chambers; the second division comprises the omasum and intestines than do herbivores, reflecting the shorter time re-
the abomasum (true stomach). The rumen and reticulum quired to digest meat than vegetation. For example, a tad-
ACQUIRING ENERC
.........................................
pole, which is almost always herbivorous, has a longer in- The general organization of the vertebrate small intes-
testine than the adult frog, which is carnivorous. tine is shown in Figure 15-20A. The outermost layer is the
The vertebrate midgut or small intestine is typically serosa, which is the same tissue that covers the visceral or-
divided into three distinct portions. The first, rather short, gans of the abdomen. The serosa overlies an outer layer of
section is the duodenum, the lining of which secretes mu-
cus and fluids and receives secretions carried by ducts
from the liver and pancreas. Next is the jejunum, which
also secretes fluid and is involved in digestion and ab-
sorption. The most posterior section, the ileum, acts pri-
marily to absorb nutrients digested previously in the duo-
denum and jejunum, although some secretion occurs from
the ileum.
As just noted, the secretory functions of the vertebrate
duodenal epithelium are supplemented by secretions from
the liver and pancreas. The cells of the liver produce bile
salts, which are carried in the bile fluid to the duodenum
through the bile duct. Bile fluid has two important func-
tions. It emulsifies fats, and it helps neutralize acidity in-
troduced into the duodenum from the stomach. The pan- ~ircu~arkusc~e '0'4,," \\ Submucosa
creas, an important exocrine organ described in Chapter 9, , \
Intestinal epithelium
The vertebrate small intestine has adaptations at every
anatomical level, from its gross anatomy to the organelles Villus
of individual cells, all designed to amplify the surface area
' available for absorption of nutrients. In humans, the lumen
of the small intestine has a gross cylindrical surface area of
only about 0.4 m2, or about 7- 8 pages of this book. How-
ever, because of the enormous elaboration of absorptive
surfaces provided by this hierarchy of structures, the true
area is increased at least 500 times, to a total of 200 to
300m2, or about the size of a doubles tennis court. Since the
rate of absorption is generally proportional to the area of Central lacteal
the apical surface membrane of the cells lining the epithe-
Figure 15-20 The anatomy of the small intestine is dominated by spe-
lium, this huge increase in surface area greatly aids absorp- cializationsfor increased surface area. (A) Overall plan. (B) Intestinalfolds
tion of digested substances from the fluid within the intes- of the mucosa are covered by (C)finger-like villi. [From "The Lining of the
tine. We will now examine this remarkable system of Small Intestine," by F. Moog. Copyright 0 1981 by Scientific American,
valleys and peaks, peninsulas and inlets. Inc. All rights resewed.]
642 I N T E G R A T I O N OF PHYSIOLOGICAL SYSTEMS
......................................
longitudinal smooth muscle, while an inner layer of circu- steadily migrate toward its tip, where they are sloughed off
lar smooth muscle surrounds the epithelial layel; which con- at the rate of about 2 x 101° cells per day in the human in-
sists of the submucosa (a layer of fibrous connective tissue) testine, meaning that the entire midgut lining is replaced
and the mucosa (or mucous membrane). Projecting into every few days.
and encircling the lumen of the small intestine are numer- The next level in the hierarchy of absorptive adapta-
ous folds of the mucosa, variously called the folds of Ker- tions is found at the apical surface of each absorptive cell,
ckring or the circular folds (Figure 15-20A and B). In addi- where there are striated structures called microvilli, which
tion to increasing surface area, these folds serve to slow the collectively form the brush border (Figure
- 15-21B and D).
progress of food through the intestine, allowing more time There are up to several thousand microvilli per cell (about
for digestion. At the next anatomical level are the finger-like 2 X lo5 per square millimeter); each standing 0.5 to
villi (Figure 15-20B and C), which line the folds, standing 1.5 pm tall and about 0.1 pm wide. The membrane of the
about 1 mm tall. Each villus sits in a circular depression microvillus is continuous with the plasma membrane of the
known as the crypt of Lieberkiihn (see Figure 15-20C). epithelium and contains actin filaments that form cross-
Within each villus is a network of blood vessels-arterioles, bridge links with myosin filaments present at the base of
capillaries and venules-and a network of lymph vessels, each microvillus (Figure 15-21C). Intermittent actin-
the largest of which is the central lacteal. Nutrients taken up myosin interaction produces rhythmic motions of the mi-
from the intestine are transferred into these blood and crovilli, which might help mix and exchange the intestinal
lymph vessels for transport to other tissues; the central chyme (semifluid mass of partially digested food) near the
lacteal can, in addition, take up larger particles. absorptive surface.
The villi are lined with the actual absorptive surface of The surfaces of the microvilli are covered by the glyco-
the small intestine, the cells of the digestive epithelium calyx, a meshwork up to 0.3 pm thick comprising acid mu-
(Figure 15-21). The epithelium consists of goblet cells in- copolysaccharidesand glycoproteins (Figure 15-21C).Wa-
terspersed among columnar absorptive cells (Figure 15-21A). ter and mucus are trapped within the interstices of the
The absorptive cells proliferate at the base of the villus and glycocalyx. The mucus is secreted by the mucous (goblet)
- Villus
-------_-___
Microvilli
Tight junction
-----I 0
Absorptive
Basal epithelial cells
D lam,ina ,
Cell membrane
u
Microvilli
- 5 ~m
Figure 15-21 The lining ofthe mammalian small intestine has a complex tions ot the suriace membrane, enclosing bundles of actin filaments. (D)
microanatomyspecializedfor absorption and secretion.The luminal sur- Scanning electron micrograph of a group of absorptive cells from the hu-
face is shown in color. (A)Avillus covered with the mucosal epithelium, man small intestine, showing the brush border. [PartsA-CfromnThe Lin-
which consists primarily of absorptive cells and occasional goblet cells. ing of the Small Intestme," by F. Moog. Copyright 0 1981
(B) An absorptive cell. The luminal, or apical, surface of the absorptive cell by Scientific American, Inc. All rights resewed. Part D from Lodish
bears a brush border of microv~lli.(C) The microvilli consist of evagina- et al., 1995.1
A Hindgut (colon) fermenter B Hindgut (cecal) fermenter Figure 15-22 The digestive tract of a
Esophagus Stomach Esophagus Stomach colon fermenter has an enlarged
colon compared with that of a cecal
fermenter, which has an enlarged ce-
cum. (A) Digestive tract of the horse
Equus caballus, a colon fermenter. Site
of fermentation isshown in red. (B) Di-
gestive tract of the rabbit Oryctolagus
cuniculus, a cecal fermenter. [Adapted
from Stevens, 1988.1
H Rectum 1 Rectum U
20 cm 10 crn
Myenteric plexus
Submucous plexus
Muscularis mucosa
Control of Motility
The coordinated contractions of circular and longitudinal
smooth muscle layers that provide alimentary canal motil-
ity in vertebrates are regulated by a combination of distinct
mechanisms.
Intrinsic control
The smooth muscle tissue in the wall of the alimentary tract
is myogenic-that is, capable of producing an intrinsic cy-
cle of electrical activity that leads to muscle contraction
without external neural stimulation. This cycle occurs as
rhythmic depolarizations and repolarizations called the ba-
sic electric rhythm (BER).This rhythm consists of sponta-
neous slow waves of depolarization that progress slowly
Figure 15-24 Coordinated contraction of the gastrointestinaltract pro-
along the muscle layers (Figure 15-25).Some of these slow
pels material through its lumen. (A) Peristalsis occurs as a traveling wave waves give rise to action potentials (APs) produced by an
of contraction of circular muscle preceded by relaxation.This produces inward current carried by calcium ions. These calcium
longitudinal movement of the bolus. (6)Segmentation occurs as alter- "spikes" lead to contractions of the smooth muscle cells in
nating relaxat~onsand contractions, primarily of circular muscle. The re-
which they occur. The amplitude of the slow-wave BER is
sult is a kneading and mixing of the intestinal contents.
modulated by local influences such as stretching of the mus-
cle tissue. Such stretching would occur when a chamber of
moving the luminal contents back into the buccal cavity. the alimentary canal is stretched by contents in its lumen.
Ruminants regularly use regurgitation to bring up the Another influence on contraction is chemical stimulation of
unchewed food for further chewing, and other vertebrates the mucosa by substances in the chyme.
use it during emesis (vomiting).
Normal peristalsis in the vertebrate stomach occurs Extrinsic (neural, hormonal) control
with the ring of contraction only partially closed. Conse- Intrinsic patterns of the BER are modulated by locally re-
quently there is a mixing action in which the contents are leased gastrointestinal peptide hormones (Table 15-1;also,
Calcium action ,
potentials
0
Smooth g
Figure 15-25 Electrical and mechanical (contraction)activity are coordi- potentials at their peaks. (B) Calcium action potentials elicit contractions
nated in the cat jejunum. (A)The slow basic electric rhythm evident as 0s- of the smooth muscle in which they occur. [Adapted from Bortoff, 1985.1
cillations in the muscle potential occasionally gives rise to calcium action
ACQUIRING ENERGY: FEEDING, DIGESTION, AND METABOLISM 647
...............................................................................
see Spotlight 9-1).Thus, a chemical stimulant in the chyme networks dispersed throughout the smooth muscle layers
can cause the release of a local hormone, and this, in turn, (Figure 15-26).The parasympathetic network made up of
can modulate the motility of the muscle tissue. cholinergic neurons is divided into the myenteric plexus
In addition to local stimuli, intestinal motility is influ- and the submucosalplexus. These plexi, which receive their
enced by diffuse innervation from the sympathetic, parasympathetic input primarily via branches of the vagus
parasympathetic, and peptidergic (purinergic) divisions of nerve, mediate excitatory actions (i.e., increased motility
the autonomic nervous system (see Chapter 9). Sympa- and gastrointestinal secretion) of the digestive tract. In con-
thetic and parasympathetic postganglionic neurons form trast, the innervation from the sympathetic division is
A
Sympathetic Longitudinal I
Circular \
Muscularis
\
Mucosa
ganglia
muscle muscle mucosae
Submucosal
Myenteric
plexus
B Vaaal plexus I
Figure 15-26 The gastro~ntesttnaltract has r~chsympathet~cand nal target tlssues (muscle, glands) are postgangl~ontc[Adapted from
parasympathet~clnnervatlon (A) Efferent sympathet~clnnervatlon Davenport, 1977 ]
(B) Parasympathettc tnnervatlon All nerve endlngs on the gastrotntestl-
648 I N T E G R A T I O N O F P H Y S I O L O G I C A L SYSTEMS
...............................................................................
A Epinephr~neadded
Tension
2 [I 0 Membrane
(9) potential
Acetylcholine added :G s
(9) 1
1: fvlez;ne
potential
Membrane
primarily inhibitory. Postganglionic neurons of the sympa- The peristaltic movements described in the previous
thetic division directly innervate all the tissues of the gut section are coordinated by the intrinsic BER, with the local
wall as well as neurons of the myenteric and submucosal participation of the myenteric plexus. This contrasts with
plexi. Activity of these sympathetic efferents inhibits the the peristaltic movements of the swallowing reflex, in
motility of the stomach and intestine. which the movements of the esophagus are under direct
The smooth muscle cells are inhibited (i.e., prevented control of the central nervous system.
from developing action potentials) by norepinephrine, Smooth muscle in the alimentary canal of vertebrates is
released by the sympathetic nerve endings, and are excited also regulated by non-adrenergic, non-cholinergicneurons
by acetylcholine (ACh),released in response to the activity of that release a variety of peptides and purine nucleotides. In
the parasympathetic nerves (Figure 15-27A). Each impulse the nearly three decades since this was first discovered,
associated with excitation produces an increment of tension, aminergic neurons have been identified that release ATP,
which subsides with cessation of impulses (Figure 15-27B). 5-HT, dopamine, GABA, while peptidergic neurons have
Evidence of the importance of the smooth muscle innerva- been found that release enkephalins, vasoactive intestinal
tion in maintaining tone is found in Hirschsprung's disease polypeptide (VIP),substance P, bombesidgastrin-releasing
(also known as congenital megacolon), in which there is a peptide, neurotensin, ch~lec~stokinin (CCK), and neu-
congenital absence of ganglion cells in the wall of the rectum. ropeptide Ylpancreatic polypeptide. This host of transmit-
Lacking smooth muscle tone, the colon becomes greatly ex- ter substances allows very fine control over the numerous
tended, which can lead to recurrent fecal impactions. interacting functions of the alimentary canal.
ACQUIRING ENERGY: FEEDING, DIGESTION, A N D METABOLISM 649
...............................................................................
TABLE 15-1
Action o f some enzymes secreted i n t h e mouth, stomach, pancreas, and small intestine o f mammals
Mouth
Salivary a-amylase Mouth Starch Disaccharides (few)
Stomach
Pepsinogen + pepsin Stomach Proteins Large peptides
Pancreas
Pancreatic a-amylase Small intestine Starch Disaccharides
Trypsinogen +trypsin Small intestine Proteins Large peptides
Chymotrypsin Small intestine Proteins Large peptides
Elastase Small intestine Elastin Large peptides
Carboxypeptidases Small intestine Large peptide Small peptides (oligopeptides)
Aminopeptidases Small intestine Large peptide Oligopeptides
Lipase Small intestine Triglycerides Monoglycerides, fatty acids, glycerol
Nucleases Small intestine Nucle~cacids Nucleotldes
Small intestine
Enterokinase Small intestine Trypsinogen Trypsin
Disaccharidases Small ~ntestine* Disaccharides Monosacchar~des
Peptidases Small intestine* Oligopeptides Amino acids
Nucleotldases Small ~ntest~ne* Nucleot~des Nucleosidases, phosphoric acid
Nucleosidases Small intestine* Nucleosides Sugars, purines, pyrimidines
*Intracellular
GASTROINTESTINAL SECRETIONS
The alimentary canal of most animals produces both en-
docrine and exocrine secretions. In fact, the alimentary
canal in many animals has been described as the "largest
endocrine and exocrine gland of the body." As explained in
Chapters 8 and 9, hormones are produced in the alimen-
tary canal by cells of endocrine glands and are liberated di-
rectly into the bloodstream, acting as messengers to recep-
tor molecules in target tissues, which usually include other
tissues of the alimentary canal.
Exocrine gastrointestinal secretions usually consist of
aqueous mixtures of substances rather than a single species
of molecule. Exocrine tissues of the alimentary canal in-
clude the salivary glands, secretory cells in the stomach and
intestinal epithelium, and secretory cells of the liver and
pancreas. The primary secretions of the exocrine glands of
the alimentary canal enter the acinar lumen of the gland,
and then generally become secondarily modified in the
gland's secretory duct. This secondary modification can in-
volve further transport of water and electrolytes into or out
of the duct to produce the final secretory juice, as illustrated
in the salivary gland (Figure 15-28)and described in detail
in Chapter 8.
Daily
Region Secretion Composition*
amount (L) PH
Buccal cavity
' Fsalivary Saliva 1+ 6.5 Amylase, bicarbonate
glands
Esophagus
Stomach Gastric juice 1-3 1.5 Pepsinogen, HCI, rennln in
infants, "lntrlnsic factor"
Gall-
Bile 1 7- 8 Fats and fatty acids, bile salts and
bladder
pigments, cholesterol
Rectum
*Exclud~ngmucus and water, which together make up some 95% of the actual secretion.
Figure 15-29 Important digestwe secretions occur a t all polnts along the human alimentary canal
The approx~matevolume and pH of each secretion IS shown on the right.
Figure 15-30 Sodium glycholate is the mam-
0 0
malian bile salt. Cholic acid (colored area) is con-
II II
C-NH-CH,-C-0- Na+ jugated with the amino acid glycine and sodium.
Glycine
Second, bile salts facilitate enzymatic fat digestion by are small enough to cross cell membranes of the intestinal
breaking down fat into microscopic droplets that collec- barrier. For example, starch, a long-chain p~l~saccharide,
tively have a much higher surface area. The ability of the is degraded to much smaller disaccharides and monosac-
bile salts to disperse fatty, water-insoluble substances de- charides; proteins are hydrolyzed into polypeptides and
rives from their amphipathic nature. That is, the bile salt then into tripeptides, dipeptides, and amino acids.
molecule contains a lipid-soluble bile acid together with a All digestive enzymes carry out hydrolysis, adding H +
water-soluble amino acid. Thus, it acts as a detergent for to one residue and OH- to the other (Figure 15-31). Hy-
the emulsification of fat droplets, dispersing them in aque- drolysis of the anhydrous bonds frees the constituent
ous solution for more effective attack by digestive enzymes. residues (e.g., monosaccharides, amino acids, monogly-
Ultimately, the bile salts are removed from the large intes- cerides) from which the polymer is formed, making them
tine by highly efficient active transport and returned to the small enough for absorption from the alimentary canal into
bloodstream. The bile salts then become bound to a plasma the circulating body fluids and for subsequent entry into
carrier protein and are returned to the liver to be recycled. cells to be metabolized.
Bile salts also disperse lipid-soluble vitamins for transport Digestive enzymes, like all enzymes, exhibit substrate
in the blood. specificity and are sensitive to temperature, pH, and certain
Third, bile fluid contains waste substances removed ions (see Chapter 3). Corresponding to the three major
from the blood by the liver, such as hemoglobin pigments, types of foodstuffs are three major groups of digestive en-
cholesterol, steroids, and drugs. These substances are either zymes: proteases, carbohydrases, and lipases.
digested or excreted in the feces.
Proteases Proteases are proteolytic enzymes, categorized as
Digestive enzymes either endopeptidases or exopeptidases. Both types of en-
An animal must first digest food before it can be used for zymes attack peptide bonds of proteins and polypeptides
tissue maintenance and growth and as a source of chemical (Figure 15-31A, Table 15-2).They differ in that endopep-
energy. Digestion is primarily a complex chemical process tidases confine their attacks to bonds well within (endo,
in which special digestive enzymes catalyze the hydrolysis "within") the protein molecule, breaking large peptide
of large foodstuff molecules into simpler compounds that chains into shorter polypeptide segments. These shorter
Peptidase ' \
C
Peptide Water Acid Amine
B
Galactose unit Glucose unit Galactose Glucose
Lactose Water
Figure 15-31 Pept~des(A)and d~sacchar~des (B) are broken down by hy- res~duesas shown, break~ngthe covalent bond holdlng the resldues
drolys~s.Under enzyme catalys~s,a molecule of water 1s added t o the two together.
TABLE 15-2
Some gastrointestinal peptide hormones
Gastrln Stomach and Secretory cells and HCl product~onand Vagus nerve actlvlty,
duodenum muscles of stomach secret~on,st~mulat~on pept~desand prote~ns
of gastric mot~l~ty in stomach
Cholecystokinin Upper small Gallbladder Contraction of
(ccK)* intestine gallbladder Fatty acids and amino
Pancreas Pancreaticjuice acids in duodenum
secretion
Secretin* Duodenum Pancreas, secretory Water and NaHCO, Food and strong acid
cells, and muscles of secretion; inhibition in stomach and small
stomach of gastric motility intestine
Gastrlc Upper small Gastrlc mucosa and lnh~b~t~
ofogastrlc
n Monosacchar~des
~nh~b~tory lntestlne musculature secretion and m o t ~ l ~ t y and fats ~n
peptlde (GIP) duodenum
Bulbogastrone Upper small Stomach lnhibition of gastric Acid in duodenum
intestine secretion and motility
Vasoact~ve Duodenum Stomach, intestine Increase of blood flow; Fats ~nduodenum
~ntest~nal secretion of thln
peptlde (VIP)* pancreat~cflu~d, rnh~blt~on
of gastrlc secretlon
Enteroglucagon Duodenum Jejunum, pancreas Inhibition of motility and Carbohydrates in
secretion duodenum
Enkephalln* Small intestine Stomach, pancreas, Stimulation of HCI Basic conditions in
intestine secretion; inhibition of stomach and
pancreatic enzyme intestine
secretion and intestinal
motility
Somastostatin* Small intestine Stomach, pancreas, Inhibition of HCI Acid in lumen of
intestines, splanchnic secretion, pancreatic stomach
arterioles secretion, intestinal
motility, and splanchic
blood flow
*Peptides marked with an asterisk are also found in central nervous tissue as neuropeptides.Neuropeptides not listed here, but identifiedin both brain
and gut tissue, include substance P, neurotensin, bombesin, insulin, pancreatic polypeptide, and ACTH.
segments provide a much greater number of sites of action sist of residues of two or three amino acids, and then fur-
for the exopeptidases. The exopeptidases attack only pep- ther break these down into individual amino acids.
tide bonds near the end (exo, "outside") of a peptide chain,
providing free amino acids, plus dipeptides and tripeptides. Carbohydrases Carbohydrasescan be divided functionally
Some proteases exhibit marked specificity for particular into polysaccharidases and glycosidases. Polysaccharidases
amino acid residues located on either side of the bonds they hydrolyze the glycosidic bonds of long-chain carbohydrates
attack. Thus, the endopeptidase trypsin attacks only those such as cellulose, glycogen, and starch. The most common
peptide bonds in which the carboxyl group is provided by polysaccharidases are the amylases, which hydrolyze all
arginine or lysine, regardless of where they occur within the but the terminal glycosidic bonds within starch and glyco-
peptide chain. The endopeptidase chymotrypsin attacks gen, producing disaccharides and oligosaccharides. The
peptide bonds containing the carboxyl groups of tyrosine, glycosidases, which occur in the glycocalyx attached to
phenylalanine, tryptophan, leucine, and methionine. the surface of the absorptive cells (see Figure 15-21C),
In mammals, protein digestion usually begins in the act on disaccharides such as sucrose, fructose, maltose,
stomach by the action of the gastric protease pepsin. There and lactose by hydrolyzing the remaining alpha-1,6 and
are different forms of this enzyme, but the most powerful alpha-1,4 glycosidic bonds. This breaks these sugars down
form functions best at a low pH value of around 2. The ac- into their constituent monosaccharides for absorption (see
tion of pepsin is aided by secretion of gastric HC1 and re- Figure 15-31B).Amylases are secreted in vertebrates by the
sults in the hydrolysis of proteins into polypeptides and salivary glands and pancreas and in small amounts by the
some free amino acids. In the mammalian intestine, several stomach, and in most invertebrates by salivary glands and
proteases produced by the pancreas continue the prote- intestinal epithelium. Many herbivores consume large
olytic process, yielding a mixture of free amino acids and amounts of plant cell walls, containing cellulose, hemicel-
small peptide chains. Finally, proteolytic enzymes inti- lulose, and lignin. Cellulose, which is in greatest abun-
mately associated with the epithelium of the intestinal wall dance, consists of glucose molecules polymerized via beta-
hydrolyze the polypeptides into oligopeptides, which con- 1,4 bonds. Cellulase, an enzyme that digests cellulose and
hemicellulose, is produced by symbiotic microorganisms in enzyme and its tissue container while it is stored in zymo-
the gut of host animals as diverse as termites and cattle, gen granules. The proenzyme is activated by the removal of
which themselves are incapable of producing cellulase. In a portion of the molecule, either by the action of another
termites, cellulase is liberated into the intestinal lumen by enzyme specific for this purpose or through a rise in ambi-
the symbiont and functions extracellularly to digest the in- ent acidity. Trypsin and chymotrypsin are good examples
gested wood. In cattle, the symbiotic microbes take up cel- of enzymes originally constituted as proenzymes. The
lulose molecules (from ingested grass, etc.), digesting them proenzyme trypsinogen, a 249-residue polypeptide, is inert
intracellularly and passing some digested fragments into the until a 6-residue segment is cleaved from the NH,-terminal
surrounding fluid. These symbiotic gut bacteria, in turn, end. This cleavage is achieved either by the action of an-
multiply and are themselves subsequently digested. Were it other trypsin molecule or by enterokinase, an intestinal
not for these symbiotic microorganisms, cellulose (the ma-, proteolytic enzyme. Trypsin also activates chymotrypsino-
jor nutritional constituent of grass, hay, and leaves) would gen through hydrolytic active form, chymotrypsin.
be unavailable as food for grazing and browsing animals.
Only a few animals, such as the shipworm Toredo (a wood- Other hgestive enzymes In addition to the major classes of
boring clam), Limnoria (an isopod), and the silverfish (an digestive enzymes just described, there are others playing
insect), can secrete cellulase without the help of symbionts. a less important role in digestion. Nucleases, nucleotidases,
and nucleosidases, as their names imply, hydrolyze nucleic
Lipases Fats are water-insoluble, which presents a special acids and their residues. Esterases hydrolyze esters, which
problem for their digestion. Fats must undergo a special, include those fruity-smelling compounds characteristic of
two-stage treatment before they can be processed in the ripe fruit. These and other minor digestive enzymes are not
aqueous contents of the digestive tract. First, fats are emul- essential for nutrition, but they enhance the efficient use of
sified-that is, they are rendered water-soluble by dispers- ingested food.
ing them into small droplets through the mechanical churn-
ing of the intestinal contents produced by segmentation (see Control of Digestive Secretions
Figure 15-24).The process of emulsification is aided by the Among vertebrates, the primary stimulus for secretion of
chemical action of detergents such as bile salts and the digestive juices in a given part of the digestive tract is the
phospholipid lecithin under conditions of neutral or alka- presence of food there or, in some instances, elsewhere in
line pH. Bile salts have a hydrophobic, fat-soluble end and the tract. The presence of food molecules stimulates chemo-
a hydrophilic, water-soluble end. Lipid attaches to the hy- sensory endings, which leads to the reflex activation of au-
drophobic end, while water attaches to the hydrophilic end, tonomic efferents that activate or inhibit motility and ex-
dispersing the fat in the water-based fluid of the digestive ocrine secretion. Appropriate food molecules also directly
tract. The overall effect is comparable to making mayon- stimulate epithelial endocrine cells by contact with their re-
naise, in which salad oil is dispersed in vinegar and egg ceptors, causing reflex secretion of gastrointestinal hor-
yolk. mones into the local circulation. These reflexes permit se-
The second step, in vertebrates, is the formation of mi- cretory organs outside the alimentary tract proper (the liver
celles (seeFigure 2-16), aided by bile salts. Micelles are ex- and pancreas, for example)to be properly coordinated with
ceedingly small spherical structures formed from molecules the need for digestion of food passing along the digestive
which have polar hydrophilic groups at one end and non- tract. Gastrointestinal secretion is largely under the control
polar hydrophobic groups at the other end and which are of gastrointestinal peptide hormones secreted by endocrine
assembled so that their polar ends face outward into the cells of the gastric and intestinal mucosa. Several of these
aqueous solution. The lipid core of each micelle is about hormones turn out to be identical with neuropeptides that
times the size of the original emulsified fat droplets, act as transmitters in the central nervous system. This sug-
greatly increasing the surface area available for pancreatic gests that the genetic machinery for producing these bio-
lipase digestion. Enzymatic degradation then results from logically active peptides has been put to use by cells of both
the action of intestinal lipases (in invertebrates) or pancre- the central nervous systerl~and the gastrointestinal tract.
atic lipases (in vertebrates), producing fatty acids plus Some gastrointestinal hormones are listed in Table 15-2.
monoglycerides and diglycerides. In the absence of suffi- Often ignored in the control of digestive secretion in an-
cient bile salts, fat digestion by the lipase is incomplete, and imals is the role of cognition or thought processes. Cephalic
undigested fat is allowed to enter the colon. influences such as mental images of food as well as learned
behaviors also stimulate digestive secretion, at least in
Proenzymes Certain digestive enzymes, in particular pro- mammals (Spotlight 15-1).However, none of these neural
teolytic enzymes, are synthesized, stored, and released in an and hormonal mechanisms regulating secretion is under
inactive molecular form known as a proenzyme, or zymo- simple voluntary control.
gen. Proenzymes require activation, usually by hydrochlo- The characteristics of digestive secretion (rate of secre-
ric acid in the lumen of the gastric gland, before they can tion, quantity of secretion) depends on several interacting
carry out their degradative functions. Initial packaging of features, including: (1)whether secretion is neurally or hor-
the enzyme in an inactive form prevents self-dgestion of the monally controlled, (2) where in the alimentary canal
trolled by the brain). Thus, in vertebrates, neural control of
digestive secretions consists of two categories. In the first, secre-
tomotor output to gland tissue occurs by an unconditioned re-
flex elicited directly by food in contact with chemoreceptors.
CONDITIONING IN In the second, secretomotor output is evoked indirectly by
FEEDING AND DIGESTION association of a conditioned stimulus with an unconditioned
stimulus.
Another example of cephalic control of secretions is the re-
The experiments of the Russian physiologist Ivan Pavlov figure
flexive secretion of salivary and gastricfluids evoked by the sight,
prominently in the histories of both psychology and physiology.
smell, or anticipation of food. This reaction is based on past ex-
Pavlov, nearly a century ago, demonstrated reflexive secretion of
perience (i.e., associative learning). Closely related to this is the
saliva in dogs. A dog was given food following the sounding of
discovery that some animals exhibit one-trial avoidance learning
a bell. Normally a dog will salivate in response to the sight or
taste of food, but not in response to a bell. However, after sev- of noxious food. Thus, a meal will be rejected even before it is
tasted if it looks or smells like something previously sampled that
eral presentations of the bell (conditioned stimulus) together
proved to be noxious. Insect-eating birds have been found to
with food (unconditioned stimulus), the bell alone elicited sali-
vation. This was the first recognition of a conditioned reflex. avoid a particular species of bad-tasting insect prey on the basis
of a one-trial experience with that prey. Examples of avoidance
These experiments became important for the development of
of noxious foods by one-trial learning have also been described
theorles of an~malbehavlor and psychology In the context of th~s
in several mammalian species.
chapter, Pavlov's experiments demonstrated that some secre-
tions of the digestlve tract are under cephalic control (~.e.,con-
secretion occurs, and (3)how long food is normally present A major secretion of the stomach lining is hydrochloric
in the region being stimulated. For example, salivary secre- acid (HCI),which is produced by the parietal, or oxyntic,
tion is very rapid and entirely under involuntary neural cells located in the gastric mucosa. The secretion of HC1 is
control, gastric secretions are under hormonal as well as stimulated by:
neural control, and intestinal secretions are slower and are
primarily under hormonal control. As in other systems, Vagal motor discharges.
neural control predominates in rapid reflexes, whereas en- The action of the gastric hormone gastrin, in conjunc-
docrine mechanisms are involved in reflexes that develop tion with histamine, a local hormone with paracrine ac-
over minutes or hours. tions synthesized in the mast cells of the gastric mucosa.
Compared with vertebrates, very little is known about (Both hormones are required for HCI secretion because
the control of digestive secretions in the invertebrates. Fil- they bind to different receptors on the parietal cell
ter feeders evidently maintain a steady secretion of digestive membrane, both of which must be filled for HCl secre-
fluids while they continuously feed. Other invertebrates se- tion to occur.)
crete enzymes in response to the presence of food in the al- Secretatgogues in food, such as caffeine, alcohol, and
imentary canal, but the precise control mechanisms have ~ ~
-
to the salivary glands. Cognitive awareness of food has an (Figure 15-32).They do this with the aid of the enzyme car-
identical effect (see Spotlight 15-1). The amylase in saliva bonic anhydrase, which catalyzes the reaction of water
mixes with the food during chewing and digests starches. with carbon dioxide:
The mucin and watery fluid conditions the food bolus to
help it slide smoothly toward the stomach by the peristaltic CO, + H20 carbon~c H2C03
movements of the esophagus. anhydrase
ACQUIRING ENERGY: FEEDING, DIGESTION, A N D METABOLISM
...............................................................................
Protein - Peptides
655
Gastric
lumen
Stomach
wall
~asoiateral Apical
Figure 15-33 The powerful proteolytic enzyme pepsin is secreted In an
membrane membrane
inactive form (pepsinogen), which is then activated by HCI. The chief
(zygomatic) cell secretes the pepsinogen, while the parietal cell secretes
Figure 15-32 The secretion of hydrochloric acid (HCI) by gastric parietal
the HCI, as well as intrinsic factor.
cells employs primary active transport of H+ produced from the break-
down of water. ACI-/HCO, exchange pump res~deson the basolateral
membrane and a C I channel on the ap~caimembrane.
ABSORPTION
The breakdown products of digestion (amino acids from
proteins, sugars from carbohydrates, etc.) are transported
from the gut to the animal's tissues and cells. In a unicellu-
lar organism, the products of digestion leave the food vac-
uole to enter the surrounding cytoplasm. In a multicellular
animal, these products must be transported across the ab- Carrier-mediated transport
sorptive epithelium into the circulation, and then move The absorption of monosaccharides and amino acids pre-
from the blood into the tissues. sents two problems. First, these molecules are hydrophilic
Digestion products are absorbed mainly via the mi- because of their -OH groups, because of charges they
crovilli of the apical membrane of the absorptive cell (see may bear, or because of both. Second, they are too large to
Figure 15-21).The digestive and absorptive mechanisms of be carried through water-filled pores by solvent drag or
the microvilli include the glycocalyx, digestive enzymes in- simple diffusion. These problems are overcome by carrier-
timately associated with the membrane, and specific in- mediated transport across the absorptive cell membrane
tramembrane transporter proteins. In the basolateral mem- (Figure 15-35). For example, sugars such as fructose are
branes other mechanisms transfer these substances out of carried down their concentration gradient by facilitated dif-
the absorptive cell into the interstitial fluid and eventually fusion, a process in which a hydrophilic, lipid-insoluble
into the general circulation. substance diffuses down its chemical gradient with the help
658 I N T E G R A T I O N O F PHYSIOLOGICAL S Y S T E M S
.........................................
Cells lining separate and non-competingcotransport systems. Each sys-
intestine tem transports just one of four categories of amino acids:
Endocytosis
Transport of sugars and amino acids across the basolateral
of specific protein channels located in the membranes. This membranes occurs by facilitated transport, as noted earlier.
process is powered by coupling sugar transport to the Some oligopeptides are taken up by absorptive cells
sodium gradients and electrical gradients across the plasma through endocytosis. In newborn mammals this process is
membrane. SGLTl is the integral membrane protein that responsible for the uptake in the intestine of immunoglob-
couples the transport of Na+ with glucose across the brush ulin molecules derived from the mother's milk that escape
border. In this model, GLUT5 is the brush border fructose digestion. Once inside the absorptive cell, nutrients pass
transporter, and GLUT2 is the basolateral membrane trans- through the basolateral membranes of the absorptive cell
porter for fructose as well as glucose and galactose (not (see Figure 15-36) into the interior of the villus and then
shown). move from the interstitial fluid into the circulatory system.
Some monosaccharides are taken up into the absorp-
tive cells by a related mechanism, hydrolase transport, in Blood Transport of Nutrients
which a glycosidase attached to the membrane hydrolyzes From the interstitial fluid of the villus, digestion prod-
the parent disaccharide (e.g., sucrose, maltose) and also ucts enter the blood or the lymphatic circulation (see
acts as, or is coupled to, the transfer mechanism of the Figure 15-36). Fishes have relatively simple lymphatic ves-
monosaccharide into the absorptive cell. sels, but these are well developed in all other vertebrates. In
Once through the epithelium, sugar and amino acid humans, about 80% of the chylomicrons, for example, en-
molecules enter the blood by diffusion into the capillaries ter the bloodstream via lymph carried in the lymphatic sys-
within the villi. Upon reaching other tissues of the body, tem, a modified ultrafiltrate of blood plasma, while the rest
sugars and amino acids are transferred by the same types of enter the blood directly. The pathway into the lymphatic
active transport and facilitated diffusion mechanisms into system begins with the blind central lacteal of the villus (see
other body cells. Figure 15-21A).In humans lymph is returned to the circu-
lation via the thoracic lymph duct. Sugars and amino acids
Active transport primarily enter the capillaries of the villus, which are
In the mammalian intestine, the sodium-driven transport of drained by venules that lead into the hepatic portal vein.
amino acids into the absorptive cells takes place via four This vein takes the blood from the intestine directly to the
ACQUIRING ENERGY: FEEDING, D l
....................................................
Figure 15-36 Lipids are transported from
-
Intestinal lumen Bile salts Fat droplet
the Intestinal lumen through absorptive cells
and into the interstitial space. Hydrolytic
Micelle products of triglyceride digestion-mono-
6 b0 +on rzr glycerides, fatty acids, and glycerol-form
1 Micelles migrate to
brush border, and
<w micelles with the blle salts in solution. Mi-
celles transport these materials to the brush
Microvillus border, where their contents enter the ab-
release their contents
\ sorptive cell by passive, lipid-soluble diffusion
across the m~crovillusmembrane Wlthin the
cell they are resynthesized into triglycerides in
the smooth endoplasmic reticulum and, to-
gether with a smaller amount of phospho-
lipids and cholesterol, are stored In the Golgi
apparatus as chylomicrons-droplets about
150 p m in diameter. These then leave the ba-
solateral portions of the cell by exocytosis.
liver. There, under the influence of insulin, much of the glu- Clearly, this quantity of water, not to mention the elec-
cose is taken up into hepatocytes, and in these cells it is con- trolytes contained within it, cannot be lost from the body
verted to glycogen granules for storage and subsequent re- with the feces. In fact, nearly all secreted water and elec-
lease into the circulation after being converted back into trolytes, along with ingested water, is recovered by uptake
glucose. The hormonal regulation of glycogen breakdown, in the intestine. Although water is reabsorbed throughout
sugar metabolism, fat metabolism, and amino acid metab- the intestine, most of the reabsorption takes place in the
olism is discussed in Chapter 9. lower part of the small intestine.
The cells in the alimentary canal responsible for water
Water and Electrolyte Balance in the Gut uptake are bound together by tight junctions near their api-
In the process of producing and secreting their various cal borders (see Figure 15-21B), nearly obliterating free
digestive juices, the exocrine tissues of the alimentary paracellular pathways. Tracer studies using deuterium ox-
canal and its accessory organs pass a great deal of water ide, D,O, indicate that water leaves the intestinal lumen
and electrolytes into the lumen of the alimentary canal. In through channels in the absorptive cell membrane that oc-
humans, this can normally amount to over 8 liters per day cupy only 0.1% of the epithelial surface. Flux studies em-
(Figure 15-37),or about 1.5 times the total blood volume. ploying isotopically labeled solutes inlcate that these
660 INTEGRATION OF PHYSIOLOGICAL SYSTEMS
.........................................
Because osmotic pressure is the motive force leading
to net water movement from the intestinal lumen to the
interior of the villus, this movement is entirely passive. In
fact, there is no evidence of active transport of water in
any living organism-animal, plant, or microbe. The
osmotic gradient driving water from the lumen into the
villus is set up primarily by the active transport of sub-
stances from the lumen into the villus, in particular the
transport of salt, sugar, and amino acids. The elevated 0s-
motic pressure within the villus that results from this active
transport, especially in the lateral clefts of the epithelium
(seeFigures 4-48 and 4-49), draws water osmotically from
the absorptive cell. This water is then replenished by wa-
ter entering osmotically across the apical membrane from
the lumen.
Most of the absorption of water and electrolytes across
the absorptive cell epithelium occurs at or near the tips of
the villi. The greater proportion of water absorption at the
villus tip results from an elevated concentration of Na+ near
the upper end of the villus lumen, which decreases with in-
creasing distance from the villus tip. There are two reasons
for this concentration gradient. First, most of the active ab-
sorption of Na+ takes place across absorptive cells located
at the tip of each villus. Chloride follows, and NaCl accu-
mulation is therefore greatest at the blind upper end of the
villus lumen. Second, the organization of circulation within
the villus leads to a further concentration of NaCl at the up-
per end of the villus lumen because of a countercurrent
mechanism (see Spotlight 12-2). Arterial blood flowing to-
ward the tip of the villus picks up Na+ and C1- from NaC1-
enriched blood leaving the villus in a descending venule.
The "short-circuiting" of NaCl in this manner recirculates
and concentrates it in the villus tip, promoting osmotic flow
of water from the intestinal lumen into the villus.
The absorption of Na+ and C1- into the villus is en-
hanced by high concentrations of glucose and certain other
hexose sugars in the intestinal lumen, which stimulate
sodium-sugar cotransport.
Excessive uptake of water from the lumen across the in-
testinal wall results in abnormally dry lumen contents (and
hence constipation). This situation is normally prevented
by an inhibitory action on electrolyte and water uptake by
some of the gastrointestinal hormones. Gastrin acts indi-
rectly to inhibit water absorption from the small intestine,
while secretin and CCK reduce the uptake of Na+, K+, and
C 1 in the upper jejunum. Bile acids and fatty acids also in-
hibit the absorption of water and electrolytes.
Unlike water, Ca2+requires a special active transport
Figure 15-37 Fluid fluxes occur all along the length of the human ali- mechanism for absorption from the gut. The calcium ion is
mentary canal. Volumes vary with the condition and body mass of the
first bound to a calcium-binding protein found in the mi-
subject. Values in black, in milliliters,are the amounts of fluid entering the
alimentary tract, and those in red are the amounts reabsorbed from the crovillus membrane and is then transported as a complex
lumen. [Adapted from Madge, 1975.1 into the absorptive cell by an energy-consuming process.
From the absorptive cell the Ca2+ then passes into the
channels exclude water-soluble molecules with molecular blood. The presence of calcium-binding protein is regulated
.
weights exceeding 200 g molk'. Smaller solute molecules by the hormone calcitriol, formerly known as 1,25-dihy-
are carried passively along with the water by solvent drag as droxy-vitamin D3. The release of Ca2+from the absorptive
it flows down its osmotic gradient through hydrated channels. cell into the blood is accelerated by parathyroid hormone.
A C Q U I R I N G ENERGY: FEEDING, D I G E S T I O N , A N D METABOLISM 661
...............................................................................
Vitamin B12, which has a molecular weight of otherwise), plus the production of body heat in birds and
.
1357 g molkl, is the largest water-soluble essential nutri- mammals. Thus,
ent taken up intact across the intestinal lumen in the region
of the distal ileum. This highly charged cobalt-containing caloric intake = caloric output
compound is associated with food protein, to which it is = calories consumed by tissues
bound as a coenzyme. In the process of absorption, B12 + heat produced
transfers from the dietary protein to a mucoprotein known
as intrinsic factor (or hemopoietic factor) that is produced Insufficient intake of calories can be temporarily offset by
by the H+-secretingparietal cells of the stomach. Since B,, using stores of fat and carbohydrates or proteins within tis-
is essential for the synthesis and maturation of red blood sues, but this produces a resultant loss of body weight.
corpuscles, pernicious anemia occurs when BIZabsorption Conversely, caloric intake in excess of what is required for
is prevented by interference with its binding to intrinsic fac- energy balance will result in increased storage of body fat,
tor. Some tapeworms "steal" B,, in the intestine of the host as in the large fat stores accumulated before long migra-
by producing a compound that removes it from intrinsic tions in migratory birds or laid down by mammals before
factor, making it unavailable to the host but available to the onset of hibernation.
tapeworm. Animals differ in their abilities to synthesize the sub-
stances fundamental to maintenance and growth. Thus, for
a given animal species, certain cofactors (Zn, I etc.) or
NUTRITIONAL REQUIREMENTS
building blocks (amino acids, etc.) essential for important
Whatever the form of food capture, ingestion, and diges- biochemical reactions or for the production of tissue mole-
tion, all animals must acquire an appropriate variety and cules may be required from food sources simply because
amount of nutritive substances, as we will now consider. those substances cannot be produced by the animal itself.
Nutrients are substances that serve as sources of metabolic Such items are known as essential nutrients.
energy and as raw material for growth, repair of tissues,
and production of gametes. Nutrients also include essential Nutrient Molecules
trace elements such as iodine, zinc, and other metals that A wide variety of molecules serve as nutrient molecules, in-
may be required in extremely small quantities. There is cluding water, proteins and amino acids, carbohydrates,
wide variation between the nutritional needs of different fats and lipids, nucleic acids, inorganic salts, and vitamins.
species. Within a species, nutritional needs vary according
to phenotypic differences in body size and composition and Water
activity, and also with age, sex, and reproductive state. A Of all the constituents of animal tissue, none is more per-
gravid (egg-bearing)or pregnant female may require more vasively important to living tissue than water. This unique
nutrients than a male, while a male producing sperm may and marvelous substance can constitute 95% or more of
have greater nutritive needs than one that is not producing the weight of some animal tissues. It is replenished in most
sperm. Regardless of reproductive state, a small animal re- animals by drinking (see Chapter 12) and by ingestion
quires more food for energy per gram of body weight than with food. Some marine and desert animals depend almost
does a larger animal, because its metabolic rate per unit entirely on "metabolic water"-water produced during
body weight is higher. Similarly, an animal with a high body the oxidation of fats and carbohydrates-to replace wa-
temperature requires more food, to satisfy greater energy ter lost by evaporation, defecation, and urination (see
needs, than does an animal with lower body temperature. Chapter 16).
(The energetics of temperature, size, and other factors are
discussed in Chapter 16). Proteins and amino acids
Proteins are used as structural components of tissues and as
Energy Balance enzymes. They can also be utilized as energy sources if first
A balanced nutritional state exists when an animal has suf- broken down to amino acids (see Chapter 3). The proteins
ficient food intake of all nutrients necessary for long-term of animal tissues are composed of about 20 different amino
growth and maintenance. The nutritional requirements in- acids. The ability to synthesize amino acids differs among
clude (1)sufficient sources of energy to power all body species. Those amino acids that cannot be synthesized by
processes, (2)enough protein and amino acids to maintain an animal, but are required for synthesis of essential pro-
a positive nitrogen balance (i.e., to avoid net loss of body teins, are the so-called essential amino acids for that animal.
proteins), (3)enough water and minerals to compensate for Recognition of this requirement has been of enormous eco-
their loss or incorporation into body tissues, and (4)those nomic significance in the poultry industry. The rate of
essential amino acids and vitamins not synthesized within growth of chickens at one time was limited by too small a
the body. proportion of a few essential amino acids in the grain diet
Energy balance requires that caloric intake over a given they were provided. Supplementing the diet with these
period of time equal the number of calories consumed for amino acids allowed full utilization of the other amino
tissue maintenance and repair and for work (metabolicand acids present in the feed, greatly increasing the rate of
protein synthesis and hence the rate of poultry growth and and fats can be converted by most animals into carbohy-
egg laying. Microbiologists artificially induce this limiting drates. The major sources of carbohydrate are the sugars,
condition by genetically engineering microbes that require starches, and cellulose found in plants and the glycogen
a specific amino acid (e.g., lysine) not normally found in stored in animal tissues.
their environment. Thus, the microbes will grow only in an
environment enriched with the amino acid, serving as a safe- Lipids
guard preventing their spread through normal populations. Lipid (fat)molecules are especially suitable as concentrated
energy reserves. Each gram of fat provides over 2 times as
Carbohydrates much caloric energy as a gram of protein or carbohydrate.
Carbohydrates are used primarily as immediate (glucose Consequently, lipids can store significantly more chemical
6-phosphate) or stored (glycogen)sources of chemical en- energy per unit volume of tissue. Fat is commonly stored by
ergy. However, they may also be converted to metabolic in- animals for periods of caloric deficit, as during hibernation,
termediates or to fats (see Chapter 3). Conversely, proteins when energy expenditure exceeds energy intake. Lipids are
TABLE 15-3
Some mammalian vitamins
Vltamln
(des~gnat~on In Major d~etary Funct~on~n Def~c~ency
letter or name) sources, solublllty* Uptake, storage mammalst symptoms
Carotene (A) Egg yolk, green or yellow Absorbed from gut with Formation of visual pig- Night blindness, skin lesions,
vegetables, fruits; FS aid of bite; stored in liver ments; maintenance of birth defects
ep~thelialstructures; im-
portant in fetal develop-
ment
Calciferol (DJ Fish oils liver; FS Absorbed from gut; little Enhancement of calcium Rickets (defective bone forma-
storage absorption from gut; tion) In children, osteomalacia
bone and tooth forma- in adults
tion
Tocopherol (E) Green leafy vegetables, Absorbed from gut; In humans, maintenance Increasedfragility of red blood
meat, milk, eggs, butter; stored in adipose and of red cells; antioxidant. cells, muscular dystrophies,
FS muscle tissues In other mammals, main- abortion, muscular wastage
tenance of pregnancy
Napthoquinone (K) Synthesis by Intestinal Absorbed from gut; little Enabling of prothrombin Failure of coagulation
flora, her, green leafy storage, excreted in synthesis by liver
vegetables; FS feces
Thram~ne(B,) Bra~n,I~ver,k~dney,heart, Absorbed from gut; Formation of cocar box- Stoppage of CHz0 metabolism
whole grains, nuts, beans, stored in liver, brain, ylase enzyme involved at pyruvate, beriberi, neuritis,
potatoes k~dney in decarboxylat~on heart failure
(Krebs cycle)
Riboflavin (B,) Milk, eggs, lean meat, Absorbed from gut; Flavoproteins in oxidative Photophobia, fissuring of the
liver, whole grains; WS stored in kidney, liver, phosphorylation skin
heart
Niacin Lean meat, liver, whole Absorbed from gut; Coenzyme in hydrogen Pellagra, skin lesions, digestive
grains; WS distributed to all tissues transport (NAD, NADP) disturbances, dementia
Cyanocobalamin Liver, kidney, brain, fish, Absorbed from gut; Nucleoprotein synthesis; Pernicious anemia, malformed
(B~d eggs, bacterial synthesis stored in liver, kidney, formation of erythrocytes erythrocytes
in gut; WS brain
Folic acid (folac~n, Meats; WS Absorbed from gut; Nucleoprotein synthesis; Failure of erythrocytes to ma-
pteroylglutamic utilized as taken in formation of erythrocytes ture, anemia
acid)
Pyridoxine (BJ Whole grains, traces in Absorbed from gut; half Coenzyme for amino and Dermatitis, nervous disorders
many foods; WS appears in urine fatty acid metabolism
Pantothenic acid Many foods; WS Absorbed from gut; Constituent of coenzyme Neuromotor, cardiovascular
stored in all tissues A (CoA) disorders
Biotin Egg yolk, tomatoes, I~ver, Absorbed from gut Protein and fatty acid Scaly dermatitis, muscle pains,
synthes~sby flora of GI synthes~s;C0,fixation; weakness
tract; WS transamination
Ascorbrc acid (C) Citrus; WS Absorbed from gut; little Vital element for collagen Scurvy (failure to form connec-
storage and ground substance; tive tissue)
antioxidant
*FS = fat-soluble, WS = water-soluble
665
666 INTEGRATlOrJ OF PHYSIOLOGICAL SYSTEMS
.........................................
an organism is one index of anabolism. That is, ana-
bolic activity leads to a net incorporation of nitrogen-
containing molecules through protein synthesis, rather
than a net loss due to protein breakdown.
Catabolism, in contrast, is the breaking down of com-
plex, energy- or material-rich molecules into simpler
ones. In caa'tabolisrn, the degradation of complex mol-
ecules into simpler ones is accompanied by the release
of chemical energy. Some of this energy is stored as
high-energy phosphate compounds, such as ATP,
which are subsequently used to power cellular aaivi-
ties (see Figure 3-70). Simpler metabolic intermediates,
such as glucose or lactate, can serve as energy storage
compounds in that they are substrates for additiona!
exergonic reactions (see Figure 3-39).
Time (min)
Figure 16-2 An oxygen deficit develops during a period of sustained, in- gradually subsides with time. The initial oxygen deficit is due to the
tense activity. Active muscle tissue with anaerobic capabilities can amass use of preexisting stores of high-energy phosphagens amassed during
an oxygen deficit, which is subsequently paid off in the form of rest. Replenishmentof these stores is included in the repayment of the
a delayed oxidation of an anaerobic product such as lactic acid. As a re- oxygen debt.
sult, an elevated metabolic rate continues after cessation of activity but
this latent heat, the mass of the water vapor is determined
by passing the air through sulfuric acid, which absorbs the
water. The energy content of each gram of water absorbed
is 2.45 kJ (0.585 kcal), the latent heat of vaporization of
water at 20°C. The results are generally reported in calories
or kilocalories per hour (see Spotlight 16-1for a discussion
of the numerous, and sometimes confusing, ways in which
energy units are expressed).
Although simple in principle, direct calorimetry can be
Direct Calorimetry rather cumbersome in practice. The technique may be too
If no physical work were being performed and chemical imprecise for animals having very low metabolic rates, and
synthesis were occurring, all the chemical energy released very large animals require a calorimetry chamber of im-
by an animal in carrying out its metabolic functions would practical dimensions. Consequently, direct calorimetry has
ultimately flow from the animal as heat. This is formalized been used most often for birds and small mammals with
in Hess's law (1840),which states that the total energy re- high metabolic rates. Another disadvantage of direct
leased in the breakdown of a fuel to a given set of end prod- calorimetry is that an animal's behavior (and therefore its
ucts is always the same, irrespective of the intermediate metabolism) is unavoidably altered because of the restric-
chemical steps or pathways used. The metabolic rate of an tions imposed by the measurement conditions.
organism is therefore effectively determined by measuring
the amount of energy released as heat over a given period. Indirect Calorimetry: Measurement from Food
Such measurements are made in a calorimeter, and the Intake and Waste Excretion
method is called direct calorimetry. The animal, usually un- Metabolic rate can be estimated from a "balance sheet"
restrained and minimally disturbed, is placed in a well-in- that sums energy gain and compares it with energy loss.
sulated chamber for the measurement. Heat lost from the Living organisms obey the laws of energy conservation and
animal is determined from the rise in temperature of a transformations that were initially derived for nonliving
known mass of water used to trap that heat. The earliest chemical and physical systems (Chapter 3). We could,
and simplest calorimeter was that devised in the 1780s by therefore, in principle determine the metabolic rate of
Antoine Lavoisier and Pierre de Laplace, in which the heat an animal in an energy steady state by using the following
given off by an animal in a chamber melted ice packed formulation:
around that chamber. The heat loss was calculated from the
mass of collected water and the latent heat of melting ice. rate of chemical energy intake
In one type of modern calorimeter, water flows through - rate of chemical energy loss
coiled copper pipes in the measuring chamber. The total = metabolic rate (heat production) (16-1)
heat lost by the subject is the sum of the heat gained by the
water plus the latent heat present in the water vapor of the Total energy intake over a given period equals the chemi-
expired air and of evaporated skin moisture. To measure cal energy content of ingested food over that same period.
SPOTLIGHT 16-1 arlsen over the use of the popular term Calone (note the capltal
C), whlch designates 1000 calorles (cal). In fact, if the label of a
ENERGY UNITS (OR soda can states that ~tconta~ns"125 CaIor~es,"almost ~nvar~ably
thls means that ~tcontalns 125 k~localor~esThe use of calorle and
WHEN IS A CALORIE kllocalorle have persisted largely because they are famll~arto
% NOT A CALORIE?) most people.
According to the lnternatlonal System of Unlts (Systeme
lnternatlonal d'unltes, SI), heat IS deflned In terms of work, and
The most commonly used unlt of measurement of heat 1s the t measurement IS the joule (J) Agaln, the more use-
the u n ~of
calone, abbreviated "cal," whlch IS deflned as the quantlty of ful verslon 1s the k~lojoule( I kJ = 1000 J) Thus, 1 cal = 4 184 J,
heat requlred to ralse the temperature of 1 g of water 1°C This and 1 kcal = 4 184 kJ If we assume a respiratory quotlent (R), -
heat quantity varies slightly with temperature, and so the calorie of 0.79, which is a typical value, 1 liter of oxygen used in the
is more precisely the amount of heat required to raise the tem- oxidation of substrate will release 4.8 kcal, or 20.1 kJ, of heat
perature of 1 g of water from 14.SQCto 15.5'C. energy.
Because a calorie is a very small quantity of heat relative to Power is the amount of energy expended per unit time, and
many biological processes, a more practical unit of heat energy has the SI units of watts(W), with 1 W = 1 J-s-'. Conversion ta-
is the kilocalorie(1 kcal = 1000cal). Unfortunately, confusion has bles for units of energy are given in Appendix 3.
Energy loss is the unabsorbed chemical energy that remains successive determinationsof the decreasing amount of oxy-
in the feces and urine produced by the animal over the same gen dissolved in the water or present in the air contained in
period. The energy content of either food or wastes can be the chamber. These measurements can be obtained very
obtained from the heat of combustion of these materials in conveniently with the aid of an oxygen electrode and the
a bomb calorimeter. In this method, the material to be tested appropriate electronic circuitry. The partial pressure of
is first dried and then placed inside an ignition chamber en- oxygen of the water or air directly determines the signal
veloped in a jacket containing a known amount of water. produced in the electrode. In gaseous phase (that is, an air-
The material is burned to ash (usingno additional fuel)with filled respirometer), 0, can be measured by a mass spec-
the aid of oxygen gas. The resulting heat is captured in the trometer or an electrochemical cell in addition to an oxy-
surrounding water jacket. The amount of energy released gen electrode. In water or air, C0, can be determined with
from the burning of the test substance is then determined a CO, electrode, but the complex chemistry of CO, dis-
from the increase in temperature of the surrounding water. solved in water (see Chapter 13) makes the interpretation
The energy released from the burned material is equivalent of these values more complex. In gases, CO, can be accu-
to that which would be released if all of this material were rately measured with a C 0 2electrode, an infrared device, a
to be passed through aerobic metabolic pathways. gas chromatograph, or a mass spectrometer. Generally, 0,
When using the balance-sheet approach to energy me- is easier to measure than CO,, so MO2is more commonly
tabolism, one must contend with variables that are difficult reported thanM,,, as a measure of metabolic rate.
to control. For example, not all the energy extracted from All of these methods of gas analysis allow mass-flow
food is available for the metabolic needs of the animal. De- analytical techniques in which the flow of gas or water into
pending on the food type, a variable fraction may actually and out of a chamber is monitored, and the difference in
be digested and absorbed in the digestive tract (Chapter gas concentrations or partial pressures is used to calculate
15).One must correct for this fraction when calculating to- respiratory exchange. Such systems employ flow-through
tal energy intake. Another complicating factor is that en- or open respirometry. Importantly,.the chambers in which
ergy can be obtained during the period of measurement the animals reside must be well stirred so that the gas exit-
from an animal's tissue reserves (e.g., stored fat). Because ing the chamber is in equilibrium with that throughout the
these energy reserves can be exhausted, the animal will chamber. Open system respirometry can also be carried out
eventually lose weight, signaling a nonsteady state (a viola- on animals fitted with breathing masks, a method especially
tion of one of the assumptions of this technique). useful in wind-tunnel tests of flying animals or on animals
The balance-sheet method does not measure BMR, running on treadmills.
SMR, or RMR (restingmetabolic rate in a fed, thermoreg- Closed and open respirometry can be combined in a sin-
ulating, inactive animal),which are best measured by more gle experiment, as illustrated in Figure 16-3. Such combined
direct methods, considered next. systems are frequently used in partitioning total gas ex-
change into pulmonary, branchial, and cutaneous locations
Indirect Measures of Metabolic Rate in such animals as amphibian larvae and air-breathing fishes
Indirect measures of metabolic rate depend on the mea- that simultaneously employ water and air breathing.
surement of some variable other than heat production re- The determination of metabolic rate from O2 con-
lated to energy utilization. The energy contained in food sumption rests on important assumptions:
molecules becomes available for use by an animal when
those molecules or their products are subjected to oxida- 1. The relevant chemical reactions are assumed to be aer-
tion, as described in Chapter 3. In aerobic oxidation, the obic. This assumption usually holds for most animals
amount of heat produced is related to the quantity of oxy- at rest, because energy available from anaerobic reac-
gen consumed. Thus, measurements of oxygen uptake tions is relatively minor except during vigorous activ-
(MO2)and carbon dioxide production ( M ~ , ~expressed
), as ity. However, anaerobiosis is important in animals that
moles of gas per hour, can be used to calculate metabolic live in oxygen-poor environments, as do gut parasites
rate.* Respirometry is the measurement of an animal's res- and invertebrates that dwell in deep lake-bottom
piratory exchange-that is, its M,~and Mco2.In closed muds. Oxygen consumption would be an unreliable
system respirometry, an animal is confined to a closed, wa- index of metabolic rate in such animals and would un-
ter- or air-filled chamber in which the amounts of oxygen derestimate the true metabolic rate.
consumed and carbon dioxide produced are monitored for 2. The amount of heat produced (i.e., energy released)
a given time period. Oxygen consumption is revealed by when a given volume of oxygen is consumed is as-
sumed to be constant irrespective of the metabolic sub-
strate. This assumption is not precisely true: more heat
* Oxygen uptake and carboc dioxide production are also commonly is ~roducedwhen 1 liter of 0, is used in the break-
expressed as volume of gas, Vo, and Vco,, respectively. This is less desir- down of carbohydrates than when fat or protein is the
able than expressing these values as molar amounts, which by definition
are comoletelv, inde~endentof measurement temoerature and atmo- substrate. However, the error resulting - from this as-
spheric pressure; Vo, or v,,> reported iq a paper can accurately
L .&
he converted into other units such asMo, or Mco, only if the author has
sumption is no greater than about 10%. Unfortu-
reported temperature and pressure, which is not always done. nately, it is generally difficult to precisely identify the
670 INTEGRATION OF PHYSIOLOGICAL SYSTEMS
...............................................................................
Figure 16-3 Open and closed respirometery
can be combined in a single experiment to
i
i measure an animal's gas exchange partition-
ing between various sites. In this experiment
on the reedfish Calamoicthys calabaricus, two
independent open systems are used to deter-
mine branchial and cutaneous aquatic oxygen
uptake. A third, closed respirometery system
Air- includes the air in the funnel above the head,
equilibrate from which the animal breathes air. Gas sam-
water ples taken after an air breath are used to cal-
culate aerial oxygen consumption. [Adapted
from Sacca and Burggren, 1982.1
substrate(s)being oxidized in correcting for differences is that it can be employed on intact, unrestrained, normally
in caloric yield. behaving animals. The many studies by Ken Nagy and his
3. The 0, stores in the body are small, so the minute-to- colleagues have shown the usefulness of this technique in
minute oxygen consumption from air or water flowing measuring field metabolic rate.
over the gas-exchange organs fairly accurately repre-
sents the metabolic rate. (Note that the ability to store Respiratory Quotient
CO, in body tissues is much greater than the ability to To translate the amount of oxygen consumed into equiva-
store 0,, so the minute-to-minute elimination of CO, lent heat production, we must know the relative amounts
is a much less reliable indicator of metabolic rate.) of carbon and hydrogen oxidized. The oxidation of hydro-
A final important method for measuring metabolic rate gen atoms is hard to determine, however, because meta-
employs isotopic techniques. These techniques first came to bolic water (i.e., that produced by oxidation of hydrogen
prominence in the measurement of water fluxes in animals: atoms available in foodstuffs),together with other water, is
deuterium- or tritium-labeled water is injected into an ani- lost in the urine and from a variety of body surfaces at a
mal, and the specific activity in serial blood or other body rate that is irregular and determined by unrelated factors
fluid samples is determined. The decline in specific activity (e.g., osmotic stress and ambient relative humidity). It is
with time indicates the loss of labeled water and therefore more practical to measure, along with the 0, consumed,
the outward water flux. The use of isotopes was then ex- the amount of carbon converted into CO,, as explained
tended to the measurement of CO, production as a mea- earlier. As noted in Chapter 13, the ratio of the volume of
sure of metabolic rate. In essence, isotopes of oxygen and CO, produced to the volume of 0, removed from within
hydrogen are injected into an animal. The subsequent de- a given time is called the respiratory quotient (RQ):
cline in the abount of isotopic oxygen ("0)in body water
is related to the rates of CO, loss through exhalation and [rate of C 0 2 production]
R - (16-2)
through water loss, the latter measured by the disappear- Q - [rate of O2 consumption]
ance of deuterium- or tritium-labeled water. Although the
numerous assumptions required must be validated for each Under resting, steady-state conditions, the RQ in Table
experimental setting, the great advantage of the technique 16-1 is characteristic of the type of molecule catabolized
TABLE 16-1
Heat production and respiratory quotient for the three major foodstuff types
Figure 16-5 Mass-specific metabolic rate (MR) in mammals declines as body mass. (6) Generalizedrelationsbetween overall metabolic rate and
body mass (M) Increases. (A) The mouse-to-elephant curve, with rneta- body mass (blackcurve)and between metabolic intensity and body mass
bolic intensity (mass-specificmetabolic rate) given as O2consumption (red curve). (C) Log-log plots of part B. The MR and log MR plots in parts
per unit mass plotted against body mass. Note the logarithmic scale of B andccross at M - 1 kg. [Part Aadaptedfrorn Schmidt-Nielsen, 1975.1
The inverse relation between metabolic rate and body line (and differs between species), and 6 is an empirically
mass applies within species as well as between species. determined exponent that expresses the rate of change of
Thus, a small human being, cockroach, or fish tends to MR with change in body mass.
have a higher metabolic rate per unit mass per unit time Mass-specific metabolic rate, also termed metabolic in-
than does a larger member of that species. However, this re- tensity, is the metabolic rate of a unit mass of tissue (i.e.,
lation is often difficult to demonstrate within a species, amount of 0, consumed per kilogram per hour). It is de-
where the overall range of body mass may be quite small termined by dividing both sides of equation 16-3 by M:
compared with that between species and where other fac-
tors such as sex, nutrition, and season may exert com-
pounding effects.
Metabolic rate is a power function of body mass, as de-
scribed by the simple relation
The relation described by equation 16-3 is shown in
color in Figure 16-5B. Because it is often more convenient
to work with straight-line rather than curved plots (e.g., for
in which MR is the basal or standard metabolic rate, M is statistical analysis) equations 16-3 and 16-4 are often put
the body mass, a is the intercept of the log-log regression into their logarithmic form. Thus, equation 16-3 becomes
674 INTEGRATION OF PHYSIOLOGICAL SYSTEMS
............................................................................
log MR = log a + b(1og M) (16-5) hypothesis. Rubner reasoned that the metabolic rate of
birds and mammals that maintain a more or less constant
body temperature should be proportional to body surface
and equation 16-4 becomes
area because the rate of heat transfer between two com-
partments (i.e., warm animal body and cool environment)
log MR
M
= log a + (b - 1)log M (16-6) is proportional, all else being equal, to their area of mutual
contact (Spotlight 16-2).The surface area of an object of
isometric shape (i.e., nonvarying proportions) and uniform
These logarithmic equations are plotted in Figure 16-5C. density varies as the 0.67 (or f ) power of its mass. This is
See Appendix 2 for a discussion of logarithmic equations. because mass increases as the cube of linear dimension,
Notice the difference in how whole-animal metabolic whereas surface area increases only as the square. As noted,
rate (black plots) and mass-specific metabolic rate (red this relation holds true for a series of animals of different
plots) change with changes in body mass. These graphs mass only if body proportions remain constant. This pro-
show that overall metabolic rate rises with increasing body vision is generally satisfied only by adult individuals of dif-
mass, whereas the mass-specific metabolic rate (metabolic ferent size within a species, because they tend to obey the
rate of a unit mass of tissue) decreases with increasing body principle of isometry -namely, proportionality of shape
mass. This principle first emerges in the mouse-to-elephant regardless of size. In this case, it follows that the surface
plot in Figure 16-SA. area must vary as the 0.67 power of body mass. However,
The value of exponent b lies close to 0.75 for many the principle of isometry is not followed in individuals of
different taxonomic groups of vertebrates and inverte-
~ -
different size belonging to related but different species. In-
brates and even holds true for various unicellular taxa stead, they tend to follow the principle of allometry-
(Figure 16-6).The exponential relation between body size namely, systematic changes in body proportions with in-
and metabolic rate has been attracting the attention of creasing species size. An example of allometry was alluded
physiologists since it was first recognized more than a cen- to earlier when we compared the proportions of an ele-
tury ago. There have been many attempts at giving a ratio- phant with those of a mouse. In a comparison of surface-
nal explanation for this nearly universal logarithmic rela- to-mass relations in mammals of different species ranging
tion between body mass and metabolism. In 1883, Max from mice to whales, surface areas were found to be pro-
Rubner proposed an attractive theory known as the surface portional to the 0.63 power of body mass (Figure 16-7).
Figure 16-6 A wide variety of groups of animals (including unicellular or- represent slopes (exponents in the allometric equation) of 0.75. The ver-
ganisms) show the same general relation between metabolic rate and tical position of each group on the graph is related to coefficient a in
body mass. Metabolic rate is related to body mass by similar exponents equation 16-3. [From Hernrningsen, 1969.1
in all three groupsforwhich data are presented here. All three solid lines
proportional to the surface area of the individual. Recall
that isometry-and, hence a 0.67, power relating surface
area to body mass-is characteristic of adult individuals of
the same species.
In spite of the logical attractiveness of the surface hy-
pothesis, it is not without flaws. True, the difference in
metabolic intensity between large and small homeotherms
may indeed be an adaptation to the more rapid loss of heat
from a smaller animal owing to surface-to-volume rela-
tions, the small animal having more surface area per unit
mass. Nonetheless, several contradictory observations raisc
serious concerns about the surface hypothesis. First, when
metabolic rates of individuals of different species of mam-
mals are plotted against body mass, the exponent relating
M a s s (g) metabolic rate to body mass is found to be approximately
0.75 (Figure 16-8B). The 0.75 power relating metabolic
Figure 16-7 Body surface area of mammals ranging from mice to whale*
rate to body mass was first discovered by Max Kleiber
is very closely correlated with body mass. The slope of the line gives an
exponent of 0.63 rather than the 0.67 predicted by the isometric (i.e., pro-
(1932)and is often referred to as Kleiber's law. The expo-
portional) scaling. The allometric (i.e., disproportional) scaling arises from nent 0.75 is significantly higher than that predicted by the
the fact that, with increasing species size, there is a progressive relative surface hypothesis (see Figure 16-8B);recall that the surface
thickening of body structures (i.e., bones, muscles, etc.), so a large area of mammalian individuals taken from various species
species has relatively less surface area than would be predicted from iso-
Of differing size is proportional to the body mass raised to
metric scaling. Recall the relat~veproportions of mouse and elephant.
[From McMahon and Bonner, 1983.1
the 0.63 power (see Figure 16-7). Thus, in comparing dif-
ferent species, the differences in metabolic rate clearly can-
not be predicted simply on the basis of differences in body
The surface hypothesis of Rubner gained support over surface area.
the years from numerous findings that metabolic rate in an- Another flaw of the surface hypothesis arises from the
imals maintaining a constant body temperature is approx- simple observation that the metabolic rates of animals
imately proportional to body surface area. An especially whose body temperatures vary with that of their sur-
close correlation can be seen in comparing the metabolic roundings (such as fish, amphibians, reptiles and most in-
rates of adult guinea pigs (all of the same species), which vertebrates) exhibit nearly the same relation to body mass
were found to be proportional to body mass raised to the as the metabolic rates of animals that actively maintain a
0.67 power (Figure 16-8A) or, assuming isometry of shape, constant, high body temperature (i.e., birds and mammals;
Elephant /
Figure 16-8 Basal metabolic rate of animals is closely correlated with species plotted against body mass. The black line through the points has
body mass. (A) Basal metabolic rate of individuals of the same species of a slope of 0.75. The red line has the slope of 0.63 predicted by the surface
various sizes plotted against individual body mass. The slope of the line law. The discrepancy is statistically significant, indicating that metabolic
indicates that the basal metabolic rate is proportional to the body mass rate is not strictly related to surface area in mammals. [Part Afrom Wilkie,
raised to the 0.67 power. (B) Basal metabolic rate of animals of different 1977; black line in part B from Kleiber, 1932.1
676 I N T E G R A T I O N OF P H Y S I O L O G I C A L SYSTEMS
.......................................
SPOTLIGHT 1 6 - 2 The velocity at which turbulence appears is higher for a stream-
lined object such as a dolphin than for an unstreamlined object
THE REYNOLDS NUMBER: such as a human scuba diver with protruding tanks, and so forth.
Because the value of a streamlined form is in reducing turbu-
IMPLICATIONS FOR BIG lence, it has no advantage for small organisms operating at very
AND SMALL ANIMALS low Reynolds numbers, in that such organisms do not experi-
ence turbulence.
To a small organism such as a bacterium, spermatozoan, or
The energy expended in propelling an animal through a fluid ciliate, the watery medium appears far more viscous than it does
medium (water or air) depends in part on the flow pattern set up to a human being. The viscosity encountered by a paramecium
in the medium. The flow pattern is determined not only by the swimming through water has been compared to the viscosity
density and viscosity of the medium, but also by the dimensions that would be experienced by a person swimming through a
and velocity of the animal. Osborn Reynolds combined these pool of honey (which, indeed, is difficult to imagine). This is an-
four factors in a dimensionless ratio that relates internal forces other example of an allometric scaling effect. Viscous effects are
(proportional to density, size, and velocity) to viscous forces. This proportional to surface area, which rises with the square of ani-
is the so-called Reynolds number (Re), calculated as mal length. Inertial effects, due to momentum of the moving an-
imal, are proportional to mass, which rises with the cube of
length. Thus, the movements of small organisms are dominated
by viscous affects, whereas those of large animals are dominated
where p is the density of the medium, V is the velocity of the by inertial effects.
body, L is an appropriate linear dimension, and p is the viscos- The relative importance of these two factors in the "coast-
ity ofthe medium. Thus, when a body moves through a fluid such ing" of objects of different size can be illustrated by forcing a tiny
as water or air, the flow pattern depends on its Re. The larger the toothpick (low Re) floating on water t o a given velocity, say
object or the higher its speed in water, the higher is its Re. The 0.1 m .s-', and then doing the same to a large floating log (high
same object moving at the same speed in air as in water would Re) of similar physical proportions but larger size. When it is let
be characterized by a lower Re in air (about 15 times lower) be- go, the little toothpick abruptly comes to rest due to the drag ex-
cause of the much lower density of air. erted on it by the viscosity and cohesion of the water. In contrast,
An Re below about 1.0characterizes movement in which the the massive log coasts for many seconds after its release be-
object produces a purely laminar pattern of flow in the fluid pass- cause its far greater momentum (based on its mass) overcomes
ing over its surface. If Re is above about 40, turbulence begins to the drag (based on its surface area). Similarly, a paramecium
appear in the wake of the object. As Re exceeds about lo6, the comes to an abrupt halt if it stops its rap~dlybeating cilia,
fluid in contact with the object becomes turbulent. At this point, whereas a whale coasts with little loss of velocity between the
the energy needed to increase the velocity further rises steeply. slow thrusts of its flukes.
see Figure 16-6). There is no self-evident reason why the ma1 size have been considered extensively. McMahon and
metabolic rate of animals with variable body temperature Bonner (1983) pointed out that the cross-sectional area
should be causally related through heat loss to body surface rather than the surface area of the body (or rather of its
area. Relatively little or no metabolic energy is expended to parts) more closely resembles the scaling of metabolic rate to
warm animals when they are in temperature equilibrium body mass, because the cross-sectionalarea of any body part
with the environment. in a series of animals of increasing size should be propor-
Scaling effects are also evident at the cellular level. tional to the 0.75 power of body mass, owing to allometric
There is a correlation between differences in metabolic in- principles that require an elephant's leg to be proportionately
tensity of animals of differing sizes and the number of mi- thicker than a mouse's leg. Remember that metabolic rate
tochondria per unit volume of tissue. The cells of a small bears the same (0.75) power relation to body mass in a wide
mammal contain more mitochondria and mitochondria1 range of animals (see Figures 16-6 and 16-8B).
enzymes in a given volume of tissue than do the cells of a Although the allometry of metabolic rate is well docu-
large mammal. Because mitochondria are sites of oxidative mented, comparative physiologists have yet to "prove" de-
respiration, this correlation comes as no surprise. However, finitively why this relation exists, and both experiments and
we are still left with the problem of how metabolic inten- thought on the subject continue. However, there is no
sity is functionally related to body size. doubt of the physiological implications of allometry to an-
The question of why large animals have lower metabolic imals. Small animals with proportionately higher metabolic
rates per volume of tissue than those of small animals and rates must spend more of their time looking for resources
the functional reasons for the allometric relations that exist and may also be more susceptible to temporary shortages
between metabolic rate (as well as other variables) and ani- of metabolic substrates or oxygen.
where MR, is the metabolic rate at the lower tempera-
ture, and MR(,+ is the metabolic rate at the higher
temperature.
The Qlo of a given enzymatic reaction depends on the
particular temperature range being considered, so it is im-
portant, when citing a Qlo value, to clearly indicate the
range of temperatures (i.e., t, and t2)for which it was de-
termined. As a rule of thumb, chemical reactions (and such
physiological
- . - -processes as metabolism, growth, locomo-
tion, etc.) have Q,, values of about 2 to 3, whereas purely
physical processes (such as diffusion) have lower tempera-
TEMPERATURE AND ANIMAL ture sensitivities (i.e., closer to 1.0).
ENERGETICS The temperature effect on enzymes results in the meta-
Few environmental factors have a larger influence on bolic rate of an animal increasing exponentially with body
animal energetics than temperature. Those animals temperature, as described by the equation
whose body temperature fluctuates with that of the envi-
ronment experience corresponding temperature-induced
changes in metabolic rate, whereas those that can main-
tain a constant body temperature in fluctuating environ-
mental temperatures have to expend metabolic energy to where MR is metabolic rate and M is body mass (making
do so. MRIM the metabolic intensity in kilocalories per kilogram
per hour), k and b, are constants, and t is temperature in
Temperature Dependence of Metabolic Rate degrees Celsius. Because enzyme action largely dictates
metabolic rate, we can see this same relation when looking
Chemical reaction rates, especially those of enzymatic re-
at the effect of body temperature on oxygen consumption
actions, are highly temperature dependent. Therefore, tis-
in animals that do not maintain body temperature at a con-
sue metabolism and, ultimately, the life of an organism de-
stant value (Figure 16-9A).Again, it is useful to transform
pend on maintenance of the internal environment at
the relation into a logarithmic one to produce a linear plot.
temperatures compatible with metabolic reactions facili-
Thus, equation 16-10 becomes
tated by enzymes. When we consider the effect of tempera-
ture on the rate of a reaction, it is useful to obtain a tem-
log MR
perature quotient by comparing the rate at two different
M
= log k + blt
temperatures. A temperature difference of 10 Celsius de-
grees has become a standard (if arbitrary) span over which
Now the coefficient b, gives the slope of the line-that is,
to determine the temperature sensitivity of a biological
the rate of increase in log MRIM per degree (Figure 16-9B).
function. The so-called Qlo is calculated by using the van't
The metabolic rates in most animals with variable body
Hoff equation:
temperature increase two- to threefold for every 10 degree
(Celsius) increase in ambient temperature, in accordance
with what would be predicted for the Qloof enzymes. Yet,
the metabolic rates of some ectotherms exhibit a remark-
where k, and k, are rates of reaction (rate constants) at able temperature independence. For example, some inter-
temperatures t, and t2, respectively. The beauty of the Qlo tidal invertebrates that experience large swings in ambient
concept is that it can be applied both to simple processes temperature with the ebb and flow of the tides have meta-
such as single enzymatic reactions and to complex bolic rates with a Qlovery close to 1.0, so the rate of me-
processes such as running and growing. To relate the van't tabolism changes very little with temperature changes as
Hoff equation to metabolic rate, consider the following large as 20 degrees. These animals appear to possess enzyme
form of the van't Hoff equation: systems with extremely broad temperature optima, which
prevents their inactivation during environmental tempera-
ture swings. Such enzyme systems may be due to a stagger-
ing of the temperature optima of sequentialenzymes in a re-
in which MR, and MR, are the metabolic rates at temper- action, such that a drop in the rate of one step in a sequence
ature t, and t2, respectively. For temperature intervals of of reactions "compensates" for an increased rate of other
precisely 10 degrees, the following simpler form of equa- steps in the sequence. The "biological clocks" of animals
tion 6-8 can be used: also are temperature insensitive with Qlosof 1.Otherwise,
the "time" kept by these "clocks" would be utterly depen-
dent on an animal's body temperature, and even a fever in
a mammal or bird would throw off its body rhythms.
678 INTEGRATION OF PHYSIOLOGICAL SYSTEMS
.........................................
a single environmental condition such as temperature. (Re-
call that evolutionary adaptation refers to evolutionary
changes over thousands of generations of a species).
Enzymatic acclimation
Acclimation occurs in individual tissues as well as in whole
animals. For instance, at a given experimental temperature,
winter acclimated frogs and summer acclimated frogs have
different contractile properties of skeletal muscles and dif-
ferent heart rates. Similarly, nerve conduction persists at
low temperatures in cold-acclimated fishes, but it is blocked
at these same temperatures in warm-acclimated ones. How
can this be explained? It is reasonable to suppose that en-
zymatic reactions have been affected. In Figure 16-10, the
plots of 0, consumption against temperature for frogs ac-
climated at 5°C and 25°C show differing slopes. That is,
- -
.-0
(1
Ambient temperature ( ' C ) exhibit different Qlos, suggesting that there has been a
modification in the temperature sensitivity of enzyme ac-
tivity. A change in the rate of enzymatically controlled re-
actions can indicate a change either in the molecular struc-
ture of one or more enzymes or in some other factor that
affects enzyme kinetics.
In some instances of acclimation, however, thermal
compensation appears to result simply from a change in the
quantity of an enzyme rather than its characteristics. This
is indicated in experiments in which the plot relating a
metabolic function to the test temperature exhibits dis-
placement without a change in slope (Figure 16-11). Be-
cause the Q,, of the process remains unchanged but the ac-
tivity is higher at every temperature in the cold-acclimated
group, the acclimation appears to have led to an increase in
the number of enzyme molecules without any change in the
Ambient temperature ( " C ) kinetics of the enzymes. The particular time course of ac-
Figure 16-9 The oxygen consumption of the tiger moth caterpillar in- climation depends on the rate at which enzyme type or con-
creases sharply as its body temperature increases. (A) Geometric coor- centration can be modified.
dinates. (B) Semilog coordinates. The generalized ordinates are shown
in color at the right in reference to equations 16-11 and 16-12. The con-
stant k is obtained by extrapolating the metabolic rate to a body tem-
perature of 0°C and is the proportionality factor in equations 16-11 and
16-12. [From Scholander et al., 1953.1
lnfrared thermal
lnfrared thermal
Scattered sunlight
- Wind
radiation from
ground
Figure 16-13 Heat IS transferredbetween an antmal and ~tsenvlronment tlon transfer heat out to the envlronment [Adapted from Porter and
In numerous ways lnfrared thermal radtatlon and d~rectand reflected Gates, 19691
sunllght transfer heat Into the anlrnal, whereas rad~atlonand evapora-
USING ENERGY: M E E T I N G ENVIRONMENTAL CHALLENGES 681
...............................................................................
whereas heat entering the body from the environment has Many animals dissipate heat by allowing water to be evap-
positive (+)value. Animals can lose heat by conduction, orated from body surfaces.
convection, radiation and evaporation. Let us now consider
each of these key terms. Heat storage Heat storage leads to an increase in temper-
ature of the heat-storing mass. The larger the mass, or the
Conduction The transfer of heat between objects and sub- higher its specific heat, the smaller its rise in temperature
stances that are in contact with each other is conduction. It (in "C) for a given quantity of heat (in joules) absorbed.
results from the direct transfer of kinetic energy of the mo- Thus, a large animal that has a small surface-to-massratio
tion from molecule to molecule, with the net flow of energy tends to heat up more slowly in response to an environ-
being from the warmer to the cooler region. The rate of mental heat load than does a small animal that has a rela-
heat transfer through a solid conductor of uniform proper- tively high surface-to-mass ratio. This follows from the sim-
ties can be expressed as ple fact that heat exchange with the environment must take
dace through the bodv surface.
0 1 2 3 4 5
Time (hours)
Figure 16-17 Microclimates under rocks afford ectotherms protec- Ambient temperature ("C)
tion from harsh thermal environments. The tropical ch~tonChiton stoke-
siican live in an intertidal zone that reaches lethally high temperatures Figure 16-18 The relation between body temperature and ambient
during the day by seeking much cooler microclimates under rocks. Tem- temperature differs in various animals. The cat is a strict homeotherm,
peratures in this illustration were recorded from the exwosed face of maintaining body temperature independent of ambient tempera-
the chiton-bearing rock, in shaded air, and in the space beneath the ture, whereas monotremes (platypus and echidna) are temporal het-
attached foot of a ch~tonhidden under a rock [Adapted from McMahon erotherms. The lizard is a strict heterotherm. [Adapted from Marshall and
e t a l , 1991 ] Hughes, 1980.1
U S I N G ENER
........................................
high daytime metabolic rate. To avoid running out of en- ing is a threat to those species living in environments with
ergy stores at night when they cannot feed, they enter into ambient temperatures below freezing. The formation of ice
a state of torpor during which they allow metabolic rate to crystals within cells is usually lethal because, as the crystals
decline and body temperature to drop toward ambient tem- grow in size, they rupture and destroy the cells. No animal
perature. Even some large endotherms resort to a long win- is known to survive complete freezing of its tissue water,
ter torpor with reduced body temperature to save energy but some come close. Certain beetles can withstand freez-
(see Hibernation and winter sleep later in this chapter). ing temperature because the extracellular fluid contains a
Regional heterotherms are generally ectotherms that substance that accelerates nucleation (the process of crystal
can achieve high core (i.e., deep-tissue) temperatures formation). Consequently, the extracellular fluids freeze
through muscular activity, while their peripheral tissues and more readily than the intracellular fluids. As ice forms in the
extremities approach the ambient temperature. As men- extracellular fluid, the unfrozen extracellular fluid becomes
tioned earlier, examples include mako sharks, tuna, and more concentrated with solutes. This process draws water
many flying insects. Elevated temperatures generally allow out of the cells, lowering the intracellular freezing point. As
higher metabolic rates than would be achieved at ambient the temperature drops further, the process continues and
environmental temperatures. Fishes that are regional het- produces further depression of the freezing point of the re-
erotherms depend on countercurrent heat exchangers. Heat maining intracellular water. The freshwater larvae of the
is conserved in the body core by a specialized parallel midge Chironomus, which survive repeated freezing, yield
arrangement of incoming arteries and outgoing veins (see some unfrozen liquid at temperatures as low as -32°C. If
Temperature relations of heterotherms) that, in the case of ice crystals form and grow within cells, they damage the tis-
heat exchangers, facilitates heat transfer between blood sue by breaking the cells. In contrast, ice crystals that form
vessels and retains heat in the body core. Some large billfin outside the cells do little damage. The adaptiveness of this
fishes (e.g., marlin) use specialized ocular muscle called process therefore lies in crystal formation in the extracellu-
"heater tissue" to elevate brain temperature (see Thermo- lar space where little tissue damage is caused. Red blood
genesis later in this chapter). Another special example of re- cells, yeast, sperm, and other cell types also can withstand
gional heterothermy is seen in the scrotums of some mam- freezing damage, provided intracellular ion concentrations
mals, including canines, cattle, and human beings, which do not rise above those levels that cause damage to cell or-
hold the testes outside the body core to keep them at a ganelles. As K. B. Storey and J. M. Storey and their col-
slightly lower temperature. The scrotum shortens in cool leagues (1992) have found, a few vertebrates, primarily
air, drawing the testes against the warmer body, and length- anuran amphibians, also can withstand freezing. Both ice-
ens as its temperature rises. These actions regulate testicu- nucleating proteins that initiate and control the formation
lar temperature and in particular prevent overheating of the of extracellular ice and cryoprotectants ("antifreezes") are
testes, which has a harmful effect on sperm production. employed to survive freezing environments.
Some animals can undergo supercooling, in which the
body fluids can be cooled to below their freezing tempera-
TEMPERATURE RELATIONS ture, yet remain unfrozen because ice crystals fail to form.
OF ECTOTHERMS Ice crystals will not form if they have no nuclei (mechanical
Ectotherms occupy a wide variety of environments- both "seeds," so to speak) to initiate crystal formation. Thus,
hot and cold. A few very specialized environments have certain fishes dwelling at the bottom of Arctic fjords live in
highly stable temperatures, varying no more than a degree a continually supercooled state and normally do not freeze.
or two throughout the year. Examples are the shallow ma- If they brush against frozen ice on the water surface, how-
rine waters under the Arctic and Antarctic ice, the deep re- ever, ice crystallizes rapidly throughout their bodies and
gions of the seas, the air deep within the interior of many they die immediately. Thus, survival for these fish depends
caves, and the microenvironments within deep groundwa- on their remaining well below the surface where ice is
ter. Normally, however, almost all environments show sig- absent.
nificant long- or short-term variation in temperature. Such The body fluids of some cold-climate ectotherms con-
variation is maximized in terrestrial temperate environ- tain antifreeze substances. For example, the body fluids of
ments, some of which may have daytime summer surface a number of arthropods, including mites and various in-
temperatures of nearly 40°C and nighttime winter temper- sects, contain glycerol, the concentration of which typically
atures of -40°C. Most animals occupy microhabitats with increases in the winter. Glycerol, acting as an antifreeze
less extreme temperature fluctuations. However, some de- solute, lowers the freezing point to as low as - 17°C. The
gree of thermal stress is inherent in living in most environ- tissues of larvae of the parasitic wasp Brachon cephi can
ments, and a wide variety of mechanisms have evolved withstand even lower temperatures; they have been super-
through natural selection to sustain animal life. cooled to -47°C without ice crystal formation. The blood
of the antarctic ice fish Trematomus contains a glycopro-
Ectotherms in Freezing and Cold Environments tein antifreeze that is from 200 to 500 times as effective as
Because the body temperature of many ectotherms depends an equivalent concentration of sodium chloride in prevent-
to a considerable extent on the ambient temperature, freez- ing ice crystal formation. The glycoprotein lowers the
temperature at which the ice crystals enlarge, but it does of denaturation. Often, critical thermal maxima relate to
not lower the temperature at which they melt. a breakdown in some critical physiological process. For ex-
For many animals living in cool (but not freezing) en- ample, most tissue functions in many ectotherms are hand-
vironments, survival requires maintaining adequate me- icapped by a decreased affinity of the respiratory pigment
tabolism at the very low levels of enzyme activity charac- for oxygen at the upper limit of body temperature. At
teristic of low temperatures. Many animals living in cold 5O0C, the blood of a chuckwalla (Sauromalus) cannot
environments have enzymes that show maximal activity at achieve more than 50% 0, saturation of arterial blood,
temperatures many degrees below those of homologous en- which prevents vigorous activity by the animal. At slightly
zymes in animals living in warmer environments. Figure lower temperatures (47-48°C) that prevent higher arterial
16-19 shows strong evidence of thermal adaptation in the oxygen saturation levels, the desert iguana Dipsosaurus
Michaelis-Menten constant (K,) of pyruvate for A,-lactate continues to be active. Above 43"C, the iguana pants, much
dehydrogenase. The Kmvalue in Termatomus centronotus, as a dog does, to increase heat loss through evaporative
a fish that lives in water that is almost always at -1.9"C, cooling.
is far higher than that in fishes and other vertebrates that in- Most ectotherms never experience such extreme tem-
habit more eurythermic environments. Even within a single peratures. Yet, even in temperate climates, many experience
species (barracuda),fish that inhabit cooler waters have a general environmental temperatures that are high enough
Kmfor pyruvate that is higher than that in individuals of the to require active responses to prevent unacceptably elevated
same species that inhabit warmer waters, allowing the body temperatures. Many ectotherms expose their bodies
cooler fish to maintain a relatively higher metabolism for to sun or to shade to absorb more or less heat, respectively,
their body temperature than that of the warmer fish. from the environment. The effectiveness of such behavioral
thermoregulation is enhanced by the high heat conductance
Ectotherms in Warm and Hot Environments of ectotherms. Certain reptiles, however, additionally use
Heat exchange with the environment is closely related to nonbehavioral (i.e., physiological) means to control the
body surface area, so the temperature of a small ectotherm rates at which their bodies heat and cool. For example,
(which has a relatively large surface area) rises and falls the diving Galapagos marine iguana Amblyrhynchus
rapidly as environmental temperatures undergo daily fluc- (Figure 14-20) can permit its body temperature to rise at
tuations. All ectotherms have a critical thermal maximum, about twice the rate at which it drops by regulating both
a temperature above which long-term survival is not possi- heart rate and flow of blood to its surface tissues. When the
ble. Generally, this is determined by measuring the temper- iguana wishes to warm up, it basks in the sun and simul-
ature at which 50% mortality occurs in a population. taneously diverts cooler core blood to the surface (see
The critical thermal maximum varies enormously, de- Figure 16-20A).The net effect is a large difference between
pending on the organism. Some thermophilic bacteria can body and environmental temperature. The increased blood
thrive at temperatures above 9O0C, although almost all flow increases the skin's heat conductance and speeds ab-
metazoans have critical thermal maxima below 45°C. The sorption of heat into the animal. Increased pumping of
physiological causes for a critical thermal maximum are blood accelerates the removal of heat from surface tissues
varied. An ultimate upper limit is the temperature at which to deeper tissues. During the iguana's prolonged feeding
proteins are denatured, though proteins such as enzymes dives in the cool ocean, loss of body heat is slowed by a re-
usually fail to function at levels well below the temperature duction in blood flow to the surface tissues and by a general
- . -----
\.**.*'-
-
Barracuda congeners:
S. idiastes (14-22°C)
S. lucasana (16-28°C)
species (barracuda).[Adapted from Sornero,
1995.1
0.00 1 I I I I I I I I I I
-5 0 5 10 15 20 25 30 35 40 45
Temperature ("C)
Hlgher body temperature,
r a p ~ dheartbeat, and
vasodilation
Figure 16-20 The Galapagos marine iguana heats and cools at different c~rculat~on, whlch qu~cklydlstrlbutes the heat throughout the body
rates, indicating an active regulation of heat exchange with its environ- Underwater heat loss 1s retarded by a slowed heartbeat and vasocon-
ments. (A) On land, the basking iguana absorbs heat from the sunk rays. strldlon In cutaneous blood vessels, both of whlch mlnlmlze the flow of
Vasodilation of cutaneous blood vessels and rapid heartbeat (as recorded blood to the skln
in the electrocardiogram, ECG) assure heating of the blood and efficient
slowing of the circulation. This is apparent in experiments birds and mammals for theirs. Indeed, the endotherms and
that demonstrate a hysteresis (asymmetrical response) of ectotherms do represent different life-styles, the former rep-
the heart rate relative to body temperature during a rise and resenting a fast, high-energy way of life and the latter rep-
fall in body temperature (Figure 16-21).Essential physical resenting a slower, low-energy approach. Many of the
concepts underlying this tactic include not only the differ-
0
ence between the rate of heat convection and the rate of
90 o Heating o
heat conduction, but also the differing heat capacities of air
Cooling
and water. Because of its far higher heat capacity, water can
remove heat by conduction from the surface of the iguana
much faster than air can; thus, it is especially important that
circulation ~othe skin be slowed during diving.
Similar dissimilarities between rates of heating and
cooling, which indicate active regulatory processes, have
been observed in amphibians and arthropods.
0 2 4
Time from onset of warmup (min)
TEMPERATURE RELATIONS
bient temperatures approaching O°C, convective heat loss is
OF HETEROTHERMS
generally so rapid that flight temperatures cannot be main-
Heterotherms are animals intermediate between pure ec- tained. High ambient temperatures, on the other hand, place
totherms and endotherms. As mentioned earlier in this the insect in danger of overheating. Thus, at ambient tem-
chapter (in Temperature classificationsof animals), certain peratures above 20°C, the hovering sphinx moth Manduca
insects and fishes are heterotherms. Some flying insects, in- . sexta prevents thoracic overheating by regulating the flow of
cluding locusts, beetles, cicadas, and arctic flies, can be con- warm blood to the abdomen. The flow of heat from the ac-
sidered both temporal and regional heterotherms because, tive thorax to the relatively inactive and poorly insulated ab-
when preparing to fly, they raise the core temperatures in domen increases the loss of heat to the environment through
their thoracic parts to more or less regulated levels. At mod- the body surface and especially through the tracheal system.
erate ambient temperatures, these insects are unable to take An interesting and somewhat unusual example of true
off and fly without prior warmup because their flight mus- shivering thermogenesis in an insect is found in honeybee
cles contract too slowly to produce sufficient power for swarms, in which individual honeybees regulate the
flight at temperatures much below 40°C. When inactive, swarm's core temperature by muscle contraction in the
however, these insects behave strictly as ectotherms. After form of shivering movements together with alterations in
such an insect is aloft, its flight muscles produce enough swarm structure. At low ambient temperatures (e.g., joC),
heat to maintain elevated muscle temperatures, and the in- the swarm packs more tightly, restricting the free flow of air
sect even employs heat-dissipating mechanisms to prevent into and out of the swarm to that needed for respiration.
overheating. These flying insects generally have quite a Through shivering activity, the core of the swarm can be
large mass; and some, such as bumblebees, butterflies, and maintained as high as 35°C. In warm weather, in contrast,
moths, are covered with heat-insulating "hairs" or scales. the swarm loosens, providing ventilatory passages for air
To warm up, these insects activate their large thoracic flight flow so that the core temperature exceeds the outside tem-
muscles, which are among the most metabolically active tis- perature by only a few degrees.
sues known. The activation is such that antagonistic mus- Another example of muscle-generated heat in a het-
cles work against one another, producing heat without erothermal species is found in the brooding female Indian
much wing movement other than small, rapid vibrations python as it elevates its body temperature with shivering
akin to shivering. Flight is finally initiated when the tho- thermogenesis so as to provide warmth for the group of
racic temperature has reached the temperature that is main- eggs around which it coils itself. In the laboratory, the rate
tained during flight, about 40°C (Figure 16-22). of occurrence of muscle contractions was found to increase
Like endotherms in general, endothermic flying insects with declining ambient temperature, and the increase in
face problems of regulating their body temperature when contractions was accompanied by an increased difference
their environmentshave large temperature gradients. At am- between the ambient and body temperatures.
690 INTEGRATION OF PHYSIOLOGICAL SYSTEMS
.......................................
Unlike terrestrial ectothermic vertebrates, which can muscle, brain, and eyes some 10 degrees or more above
bask in the sun to warm up, marine ectotherms cannot their surroundings. These fishes can therefore be classified
obtain radiant energy as a source of underwater heat, ow- as regional heterotherms. The large mass (and hence small
ing to the high infrared absorption of water. As a result, surface-to-volume ratio) of some of these fishes helps them
fishes can rise above ambient temperature only through in- attain a relatively constant muscle temperature. In these
tensive metabolic activity. Many teleosts are strictly ec- fishes, the retention of heat in the body core depends
tothermic, operating at core temperatures close to ambient crucially on the organization of the vascular system. Unlike
temperatures. However, as already mentioned, some fishes, ectothermic fishes, which have a centrally located aorta and
such as tunas, have specializations for generating and postcardinal vein (Figure 16-23A), heterothermic fishes,
retaining sufficient heat to raise the temperature of body (e.g., tunas and lamnid sharks such as the mako) have
A Ectothermic
Dorsal
B Heterothermic
cutaneous Cutaneous
artery vein
Water 19.3"
Y
C
35 - ..
.,- /
-0
0)
//
2 20-
E //
/
Figure 16-24 The bluef~n tuna controls reg~onalbody temperature w ~ t h
5E
a countercurrent arter~al-venousheat-exchange rete The rete (shown In
color) helps the tuna retaln heat produced In act~vedeep muscles (A) En- ' - //
largement of the rete area (B) Isotherms (left), plotted at 2 Cels~usdegree r" /I
Intervals, show temperature d~str~but~on In cross sectlon [rrght) (C)Max- //
lmum muscle temperatures of bluef~nscaught In waters of d~fferenttem- 10 I 1 I
peratures The dashed Ihne nd~catesequalty between body tempera- 10 15 20 25 30
tures and water temperatures [From Carey and Teal, 1966.1 Water temperature ( ' C )
Another important factor is that these regional het- man beings are a vestige of the pilomotor control of a long-
erotherms swim continuously, so the red muscle never lost pelage.
cools down to the ambient temperature. One of the As the ambient temperature decreases, an endotherm
implications of regional heterothern~yis the energy sav- will eventually reach its lower critical temperature (LCT;
ings in the cool tissues while the temperatures of only see Figure 16-25),below which the basal metabolic rate be-
certain tissues, such as the swimming muscles, are being comes insufficient to balance heat loss despite these mani-
elevated. fold adjustments in thermal conductance. Below this tem-
perature, an endotherm must increase heat production
above basal levels to offset heat loss (i.e., by thermogenesis,
TEMPERATURE RELATIONS described in the next subsection). Heat production then
OF ENDOTHERMS rises linearly with decreasing temperature below the lower
In homeothermic endotherms (most mammals and birds), critical temperature, in what is termed the zone of meta-
body temperature is closely regulated by homeostatic bolzc regulation (see Figure 16-25). If the environmental
mechanisms that regulate rates of heat production and heat temperature drops below the zone of metabolic regulation,
loss so as to maintain a relatively constant body tempera- compensating mechanisms fail, the body cools, and the
ture independent of environmental temperatures. As men- metabolic rate drops. Many animals tolerate varying de-
tioned earlier, core temperatures are maintained nearly con- grees of hypothermia during their normal rest period (in-
stant between 37°C and 38°C in mammals and about 40°C cluding human beings during sleep). However, if an ani-
in birds. The temperatures of peripheral tissues and ex- mal's body temperature falls below its normal values, the
tremities are held less constant and are sometimes allowed animal enters a state of hypothermia (see Figure 16-25).If
to approach environmental temperatures. Basal heat pro- this condition persists, the animal grows progressively
duction for different homeotherms of a given size is about cooler and, because cooling only further lowers the meta-
the same, and the basal metabolic rate can be 10 times as bolic rate, the animal soon dies.
high as the standard metabolic rate of ectotherms of com- The thermal neutral zone lies entirely below the normal
parable size measured at similar body temperatures. This body temperature, Tb(37-40°C), as shown in Figure 16-25.
elevated basal metabolism, in conjunction with heat-
conserving and heat-dissipating mechanisms, allows
Thermal neutral
homeotherms to maintain constant body temperatures zone
as much as 30 Celsius degrees or more above ambient
temperatures.
Blood flow in
Figure 16-26 Brown-fat deposits are found between the scapulae in tion of this tissue. During brown-fat oxidation, this tissue is detectable as
bats and many other mammals. The detail shows the special vasculariza- a warm region by its infrared emission.
massive heat production. This fat contains such extensive Brown fat heats up significantly during thermogenesis.
vascularization and so many mitochondria that it is brown This newly produced heat is rapidly dispersed to other
(owing largely to mitochondrial cytochrome oxidase) parts of the body by blood flowing through the extensive
rather than white. In brown fat, oxidation takes place vasculature of the brown fat tissue. Nonshivering thermo-
within the fat cells themselves, which are richly endowed genesis is especially pronounced during arousal of hiber-
with fat-metabolizing enzyme systems. In ordinary body nating or torpid mammals, when it supplements shivering
fat, deposits must first be reduced to fatty acids, which en- to facilitate rapid warming. One consequence of acclima-
ter the circulation and eventually are taken up by other tis- tion to cold by mammals is an increase in brown fat
sues, where they are oxidized. deposits, which allows for a gradual changeover from
Nonshvering thermogenesisin fat (includingbrown fat) shivering to nonshivering thermogenesis at low ambient
is activated by the sympathetic nervous system through the temperatures. The acclimatory increase in brown-fat ther-
release of norepinephrine, which binds to receptors on the mogenesis is mediated by the thyroid hormones. Brown
adipose cells of brown-fat tissue. Through a second-mes- fat is also present in some mammalian infants, including-
senger mechanism, described in Chapter 9, this signal leads human infants, where it is generally located in the region
to thermogenesis by two mechanisms. In the first of these of the neck and shoulders, the spine, and the chest. Because
mechanisms, normal ATP utilization for cellular processes an infant is relatively small and inactive at birth, deposits
rises in these fat cells in response to the sympathetic signal, of brown fat provide an important and rapid means of
accounting for part of the increased heat production. warming if the infant is threatened with temperature
Through processes such as ion pumping by the plasma reduction.
membrane, ATP is hydrolyzed to produce work and heat. In Another example of tissues specialized for heat produc-
the second mechanism, ATP production is uncoupled dur- tion are the heater tissues formed from modified eye muscles
ing respiratory chain oxidation. The resynthesis of ATP in billfishes. B. A. Block and her colleagues (1994)have ex-
from ADP and Pi is normally coupled to the movement of tensively investigated these tissues, which have an enormous
protons (H+)down their electrochemical gradient from in- capacity to generate heat (as high as 250 W . kg-'). Heater
termembrane space into mitochondria across the inner mi- cells, which lack myofibrils and sarcomeres, produce heat
tochondrial membrane. Thermogenesisin brown fat is char- through the release of Ca2+ from internal cytoplasmic
acterized by the appearance in the inner mitochondrial stores. The released Ca2+ ions then stimulate catabolic
membrane of uncoupling proteins, which provide a path- processes and mitochondrial respiration (Figure 16-27).
way for protons to leak across this membrane without the
energy of their downhill movement being harnessed for the Endothermy in cold environments
phosphorylation of ADP to ATP. Once inside the mito- Endotherms adapted to cold environments have necessar-
chondrion, the protons oxidize substrate oxygen to produce ily evolved a number of mechanisms, both temporary and
water and heat, or else further utilization of metabolic en- permanent, that help them retain body heat. For example,
ergy is required to subsequently pump them into the inter- an animal sensing heat loss in a windy place will fluff its fur
membrane space and eventually out of the mitochondria. or feathers and move to a more sheltered area. This reduces
Figure 16-27 The metabolism of heater tissues in billfish is specialized ceptor activates a sarcoplasmic reticulum (SR) Ca2+channel, releasing
for heat production. This model for nonshivering thermogenesis in the Ca2+stored in the sarcoplasmic reticulum. The increased cytoplasmic
skeletal muscle of billfish heater tissue shows how exogenously stimu- Ca2+ions then triggerthe release of energy in the form of heat from ATP
lated release of Ca2+into the cytoplasm stimulates mitochondrial respi- previously manufactured in the mitochondria. [Adapted from Block,
ration and the associated release of heat. After stimulation, a T-tubule re- 1994.1
U S I N G ENE
.......................................
convection and the dissipation of body heat by the wind.
More enduring responses to cold include the thick layers of
insulation in many arctic animals, in the form of subcuta-
neous fat or a thicker pelage or plumage. The insulating ef-
fectiveness of pelages in arctic and subarctic animals
changes with both season and latitude to match insulation
qualities with insulation needs. In addition, animals living
in the temperate zone exhibit seasonal variations by shed-
ding old fur or feathers and growing new bodily cover,
thereby providing thick insulation during the winter, yet
preventing overheating during the summer.
The specific conductances of homeotherms vary over
a large range and decrease with body size (Figure 16-28).
Ambient temperature ("C) 1,
Larger animals have lower specific heat conductances ow-
ing to their generally thicker coats of fur or feathers. In ad- Figure 16-29 The decline in metabolic rate in endotherms as ambienttem-
dition, they face smaller heat loss in cold climates because peraturesfall depends on the extent of the animal's insulation.Adecrease
of their relatively smaller surface areas. Thus, one adapta- in insulation (i.e., an increase in conductance) raises the lower critical tem-
tion of endotherms to cold latitudes is an increase in body perature and makes the slope of increasing metabolism steeper. The slope,
however, still extrapolates to body temperature at zero metabolic rate.
size. As the surface-to-volme ratio becomes smaller, pelage
becomes thicker and conductance decreases. With in-
creased insulation, the lower critical temperature of a within the body and is supplied with blood vessels. Thus,
homeotherm decreases and the thermal neutral zone ex- whereas the insulating properties of fur remain unaffected
tends to lower temperatures (Figure 16-29). An exception by circulatory adjustments to the area, the insulating prop-
is that small animals and immature animals often have erties of blubber depend on whether blood flow to the sur-
feathers or fur that is less conductive per unit thickness, as face is restricted or not. Hence, the more blood flow is di-
evident in the fluffy feathers of young chicks, for example. verted away from the vessels within the blubber, the higher
Blubber, a fatty tissue typically found under the skin in the effective thickness of the insulating layer. Conversely, the
cetaceans, is a good insulator because, like air, blubber has greater the blood flow into the blubber, the lower the effec-
a lower thermal conductivity than does water, which is the tive thickness of the insulating layer. This ability to regulate
main constituent of nonfatty tissues. In addition, fatty tis- heat transfer through blubber allows a marine mammal to
sues are metabolically very inactive and require little per- facilitate the loss of excessive body heat by shunting its sur-
fusion by blood, which would ordinarily carry heat to be face blood effectively to the outer regions of the insulating
lost at the body surface. In whales, the outermost regions layer of blubber during periods of intense activity in
of the thick blubber layer are always at a temperature near warmer waters or when lying on land in warm air.
that of the surrounding water.
An important means of controlling heat loss from the Countercurrent heat exchange
surface is the diversion of blood flow to or away from the Effective locomotion requires that the limbs of endotherms
skin (see Figure 16-14).Vasoconstriction of arterioles lead- not be mechanically hindered by a massive layer of insula-
ing to the skin keeps warm blood from perfusing cold skin tion. The flukes and flippers of cetaceans and seals and the
and conserves the heat of the body core. An interesting ad- legs of wading birds, arctic wolves, caribou, and other cold-
vantage of blubber over pelage in the control of heat loss is weather homeotherms require blood to nourish cutaneous
illustrated in Figure 16-15B, which reminds us that fur is lo- tissue and limb muscles used in locomotion. The well vas-
cated outside the body proper whereas blubber is contained cularized limbs are potential major avenues of body heat
loss because they are thin and have large surface areas.
Excessive heat loss from these appendages can be re-
duced drastically by countercurrent heat exchange. Coun-
tercurrent exchange systems have already been discussed
in the context of oxygen and carbon dioxide exchange
(Chapter 13). Arterial blood, originating in the animal's
core, is warm. Conversely, the venous blood returning from
peripheral tissues may be very cold. As blood flows from the
core, it enters arteries in the limb that lie next to veins that
carry blood returning from the extremity. As the arteries
and veins pass each other, the warm arterial blood gives up
Body mass (g)
heat to the returning venous blood and thus becomes suc-
Figure 16-28 Thermal conductance decreases exponentially as body cessively cooler as it enters the extremity. By the time it
mass Increases. reaches the periphery, the arterial blood is precooled to
696 INTEGRATION OF PHYSIOLOGICAL SYSTEMS
.........................................
within a few degrees of the ambient temperature, and little well as other large mammals, can absorb relatively large
heat is lost. Conversely, the returning venous blood is quantities of heat for a given rise in body temperature. These
warmed by the arterial blood, so it is nearly at core tem- features also result in a slow loss of heat during the cool
perature as it flows into the core. The advantage of such a hours of the night. Thus, the large mass acts as a heat buffer
system is that heat exchange with the environment is re- that, by reducing rates of both absorption and loss of heat,
stricted without a reduction in blood flow and the oxygen minimizes extreme temperature fluctuations. A dehydrated
and nutrients that the blood carries. A similar situation was camel can also tolerate an elevation of its core temperature
described for heterothermic fishes (see Figures 16-23 and by several degrees, further increasing its heat-absorbing ca-
16-24).Another highly evolved example of countercurrent pacity. Large amounts of heat gradually accumulated during
heat exchange is found in the mirabile of the flipper of the daytime hours are subsequently dissipated in the cool of the
porpoise. Here, the artery carrying warm blood flowing to- night. In preparation for the next onslaught of daytime heat,
ward the extremity is completely encased in a circlet of veins the dehydrated camel allows its core temperature to drop
carrying cold blood back from the extremity. Birds and arc- several degrees below normal during the night. As a conse-
tic land mammals also use countercurrent exchange to min- quence, the camel starts the day with a heat deficit, which al-
imize heat loss from their extremities in cold climates, and lows it to absorb an equivalent amount of additional heat
to some extent this mechanism is also present in the ex- during the hot part of the day without reaching harmful tem-
tremities of human beings. As a result, the extremities of peratures. This practice, called limited heterothermy, allows
cold-climate endotherms are maintained at temperatures the camel to tolerate the extreme daytime desert heat with-
that are far below the core temperature and often approach out using much water for evaporative cooling.
ambient temperature (Figure 16-30). The efficiency of the Limited temporal heterothermy is also practiced by the
countercurrent heat exchangers can generally be regulated antelope ground squirrel (Ammospermophilus leucurus), a
by vasomotor control in which blood flow is shunted past diurnal desert mammal. Because of its small mass, the an-
the heat exchanger network by parallel vessels. telope ground squirrel cannot continuously gain heat for
several hours in the hot sun, and its small surface-to-mass
Endothermy in hot environments-dissipation of body heat ratio would lead to rapid heating. Instead, this desert mam-
In very hot, dry climates, large animals have the advantages mal exposes itself to high environmental temperatures for
of relatively low surface-to-mass ratios and large heat ca- only about 8 minutes at a time. It then returns to its bur-
pacities. Camels, well known for their ability to tolerate heat, row, where its stored heat escapes into the cool under-
have not only a large body mass, but also a thick pelage that ground air. By allowing its temperature to drop a bit below
helps insulate them from external heat. Low surface-area-to- normal before returning to the hot desert floor, it is able to
volume ratio and thick pelage retard the absorption of heat extend its stay a few minutes without lethal overheating.
from the surroundings. Furthermore, because of its large The temperature of the body surface is an important
mass and the high specific heat of tissue watel; the camel, as factor affecting heat loss to the environment, because it de-
Air = - 16°C
Air = -31°C
--
lnspiration Expiration
Figure 16-32 The pathway of respiratorygas flow varies with the extent
Both homeothermic endotherms and homeothermic ec-
totherms also use nonmetabolic means to regulate body
temperature. The regulation of body temperature, Tb, is
not fully understood even after decades of research on the
of panting in the dog. (Top) Air flow through the nose of a panting dog. topic but appears to work along principles of negative feed-
Horizontal lines extending to the left of the vertical midline indicate in- back (see Spotlight 1-1). Most animals have not one but
spiration; to the right, expiration. Mean inhaled and exhaled volumes are
many temperature sensors in various regions of the body.
indicated by vectors placed adjacent to the dog's nose. (6ottom)Air flow
through the mouth of a panting dog. lnspirationthrough the mouth isvir- Furthermore, to maintain Tb at about T,,,, homeothermic
tually zero; expiration through the mouth carries most of the air taken in animals can call on several heat-producing and heat-ex-
through the nose. [From Schmidt-N~elsenet al., 1970.1 changing mechanisms, so the thermostat controls heat-
conserving and heat-loss mechanisms as well as heat endings exist in the brain, the spinal cord, the skin, and sites
production. This means of control is analogous to the in the body core, providing input to thermostatic centers in
microprocessor-controlled heating and cooling system of the brain. Although a mammal may have several ther-
the futuristic "smart house" in which the thermostat, in ad- moregulator~centers, the most important one, considered
dition to cycling the furnace and air conditioner, controls to be the body's "thermostat," is located in the hypo-
the position of window shades, window opening and clos- thalamus (see Figure 9-5). It was discovered by Henry G.
ing, the conductance of the wall and roof insulation, and so Barbour in 1912 in the course of a series of experiments in
forth. Furthermore, control of thermogenesis in the which a small temperature-controlledprobe was implanted
homeotherm is not all-or-none, like the turning on and off in different parts of the rabbit brain. The probe evoked
of a furnace. Instead, the rate of heat production by meta- strong thermoresponses only when it was used to heat or
bolic means is graded according to need. The colder the cool the hypothalamus. Cooling the hypothalamus pro-
temperature sensors become (within limits), the higher the duced an increase in metabolic rate and a rise in Tb,
rate of therrnogenesis. Engineers call this proportional con- whereas heating it evoked panting and a drop in Tb. This
trol, because heat production and conservation are more or experiment is analogous to changing the temperature of a
less proportional to the difference of Tb - Ts,. thermostat in your home by holding a lighted match
nearby. As the thermostat is warmed above its set-point
The hypothalamus-the mammal's "thermostat" temperature, it shuts down the furnace, allowing the room
Mammalian body temperature can vary widely (as much as temperature to drop below the set point. An apparatus for
30 Celsius degrees) between the periphery and the body controlling hypothalamic temperature and measuring a
core, with the extremities undergoing far more variation homeotherm's response to changes in that temperature is
than the core. Temperature-sensitive neurons and nerve shown in Figure 16-33.
Figure 16-33 Temperature regulation by the hypothalamus can be mea- hypothalamus.Metabolic rate and evaporative water loss are measured
sured by experimentally changing hypothalamic temperature. The ap- by analyzing the effluent air for water, O, and CO, content. The meta-
paratus measures the temperature sensitivity of the hypothalamus and bolic chamber is at constanttemperature [From "The Thermostat of Ver-
thermoregulatory response to changes in hypothalamic temperature, tebrate Animals" by H. C. Heller, L. I. Crawshaw, and H. T. Hammel. Copy-
which is altered by means of a water-perfused thermode implanted in the right 0 1978 by Scientific American, Inc. All rights resewed.]
Experimental procedures like Barbour's have shown Thus, experimentally lowering hypothalamic temperature
that the mammalian hypothalamic thermostat is highly in a dog leads to elevated metabolic heat production by
sensitive to temperature. Variation of mammalian brain shivering. On the other hand, warming the dog's hypothal-
temperature of only a few Celsius degrees seriously affects amus elicits the heat-dissipating response of panting.
the functioning of the brain, so it is not surprising to find A rise in core temperature of only 0.5 Celsius degrees in
the major thermoregulatory center of mammals located most mammals causes such extreme peripheral vasodila-
there. Neurons that are highly temperature sensitive are lo- tion that the blood flow to the skin can increase several
cated in the anterior part of the hypothalamic thermostat. times above normal. In human beings, this response pro-
Some of these neurons show a sharply defined increase in duces a flushed appearance to the skin. The effect of ele-
firing frequency with increased hypothalamic temperature vated core temperature on peripheral vasodilation and,
(Figure 16-34).These neurons are believed to activate heat- hence, skin temperature is illustrated in Figure 16-35,
dissipating responses such as vasodilation and sweating. which shows that the skin temperature of a rabbit's ear rose
Others show a decrease in firing frequency with increase in very sharply from less than 15°C to about 35°C at the point
temperature above a certain value. Still other neurons in- at which the rabbit's core temperature exceeded 39.4"C.
crease their firing frequency when the brain temperature Because the temperature of the ear reached a maximum, it
drops below the set-point temperature. They appear to con- can be assumedthat the vessels of the ear dilated fully as
trol the activation of heat-producing responses (e.g., shiv- soon as the core temperature exceeded this limit.
ering, nonshivering thermogenesis, brown-fat metabolism) The effect of the hypothalamic thermostat on such per-
and heat-conserving (e.g., pilomotor) responses. ipheral heat-exchange mechanisms is about 20 times as
In addition to the information about its own tempera- great in some mammals as reflexive adjustments initiated
ture generated by these thermosensitiveneurons, the hypo- by peripheral temperature sensors. This hypothalamic
thalamus receives neural input from thermoreceptors in "override" is significant in light of the importance of care-
other parts of the body. All this thermal information is in- fully regulated brain temperature. Without dominance of
tegrated and used to control the output of the thermostat. the hypothalamic thermostat, an internally overheated an-
Neural pathways leaving the hypothalamus make connec- imal exercising in a cold environment would fail to activate
tions with other parts of the nervous system that regulate heat-dissipating blood flow to the surface capillaries, and
heat production and heat loss. Some of these pathways are its core temperature would continue to rise to dangerous
activated by high temperatures signaled from peripheral levels.
and spinal thermoreceptors and by the hypothalamic tem- In some homeotherms, especially small animals subject
perature-sensitiveneurons. The efferent pathways activate to rapid cooling at low ambient temperatures, the set-point
increased sweating and panting, as well as a lowered pe- temperature of the hypothalamic thermostat changes with
ripheral vasomotor tone,that produces increased blood ambient temperature, presumably because ambient devia-
flow to the skin. Conversely, body cooling leads to ther- tions are sensed by peripheral receptors. Thus, in the kan-
mogenesis and increased peripheral vasomotor tone. These garoo rat, a sudden drop in ambient temperature is quickly
same responses can be elicited without cooling the whole followed by a rise in set-point temperature. This rise causes
body by simply cooling the neurons of the hypothalamus. an increase in metabolic heat production in anticipation of
increased heat loss to the environment.
t
processes (sweating or regulation of body temperature is only
through control of heat conductance to the
ing)
Intensity environment by varying the peripheral blood
flow or the insulating effectiveness of fur or
response feathers (black plot). Above and below this
range, these passive measures are exhausted,
Passive and active thermogenesis (left) or evaporative
processes heat loss (right)develops (red plots).
Figure 16-37 Hypoxla Induceschanges In selected body temperature In goldflsh (Carassfus auratus) and toads (Bufo mannus). [Adapted from Wood, 1991.]
elevation in the set point to a higher temperature than nor- laboratory conditions simulating a desert environment
mal. The result is that the body temperature rises several before and after administration of pyrogenic bacteria
degrees, and the animal experiences a fever. Anesthetics and (Figure 16-38).In response to the fever-producing bacte-
opiates such as morphine, in contrast with pyrogens, cause ria, the lizards positioned themselves more frequently
a lowering of the set-point temperature and hence a drop in in radiantly heated zones of the artificial environment,
body temperature. The adaptive significance of endogenous effectively raising their temperatures to unusually high
pyrogens and of their production of fever in homeotherms levels (i.e., fever). This behavioral response and the fever
may relate to the bacteriostatic effects of elevated body temperatures it produced conferred protection against
temperature. bacterial infection. This protection is thought to take
Pyrogenic bacteria elevate body temperature in some two forms: (1)the antiviral and antitumor agent inter-
ectotherms as well as in endotherms. In a classic experi- feron is more effective at higher temperatures, and (2)
ment by H. A. Bernheim and M. G. Kluger (1976), body elevated temperatures also diminish the growth of some
temperatures were monitored in a desert iguana under microbes.
-
9
$? 40
Day 3
Day 2
resulting
from
increased
by pyrogenlc bacteria, the llzards ralsed thelr
body temperatures above normal levels by ~ n -
creased basklng behavlor The graph shows
a
C
bask~ng the Increase In body temperature wlth succes-
$Q slve days after lnjectlon of the bacteria
[Adapted from Bernhelm and Kluger, 19761
E
C
g 38
f)
a,
a
E
0
a 36
Purogenic
34 Day 1 bacteria
injected
10 12 2 4
Time of day (hour)
USING ENE RGY: M E E T I N G E N V I R O N M E N T A L CHALLENGES 703
. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
Thevrnovegulation during activity
The energy efficiency of muscle contraction is only about
25%. For every joule of chemical energy converted into
mechanical work, 3 J of energy is degraded to heat. During
exercise, this extra heat, added to the heat produced by
basal metabolism, will cause a rise in body temperature
above the set-point temperature unless it can be dissipated
to the environment at the same rate at which it is produced.
Most of the excess heat manages to be transferred to the
environment, but a rise in core temperature of homeo-
therms does occur during exercise, indicating incomplete
removal of the excess heat. The rise in temperature is mod-
erately useful in two respects: (1)it increases the difference
Tb - Taand thereby increases the effectiveness of the heat-
loss processes by increasing the gradient for heat loss, and
(2)it leads to an increased rate of metabolic reactions, in-
cluding those that support physical activity. However, the
core temperature can rise to dangerously high levels during
heavy exercise in warm environments, and this excess heat
has to be dealt with.
The level to which the core temperature rises in home-
otherms is proportional to the rate of muscular work. Dur- Internal temperature ("C)
ing light or moderate exercise in cool environments, body
temperature rises to a new level and is regulated at that Figure 16-39 Rate of sweating In humans increasessharply as body tem-
perature approaches about 37°C. Core temperature was elevated by
level as long as the exercise continues. Thus, temperature
exercise or by elevating the ambient temperature. [Adapted from
appears to remain under the control of the body's thermo- Benzinger, 1961.1
stat. The rise in temperature proportional to the level of ex-
ercise appears to be a consequence primarily of an increase
in the error signal, Tb - T,,,, of the thermostatic feedback set of heavy physical work, even though there is no de-
control by the hypothalamus. The error signal is the differ- tectable increase in skin temperature in that time. However,
ence between the thermostat's set point and the actual core core blood temperature shows a detectable rise in temper-
temperature. The greater this difference (i.e., the greater the ature within 1 second after exercise has begun. Apparently,
error signal), the greater the activation of heat-loss mecha- the onset of sweating, nearly concurrent with the onset of
nisms. Thus, the rate of heat dissipation increases as the neural activity underlying exercise, results from the reflex
core temperature rises above the set point, and a new equi- activation of sweating by central temperature receptors.
librium becomes established between heat production and The set points for heat loss are lower in well trained ath-
heat loss. During heavy exercise, especially in warm envi- letes, especially in warm weather.
ronments, the heat-dissipating mechanisms are not able to A special countercurrent heat exchanger to prevent
balance heat production until body temperature rises sev- overheating of the brain during such strenuous exercise as
eral degrees, increasing the T, - Ta difference. Thus, ele- running is employed by certain groups of hoofed mammals
vations of 4 to S Celsius degrees in core temperature are (e.g., sheep, goats, and gazelles) and carnivores (e.g., cats
commonly observed in human beings after strenuous, sus- and dogs). This system, the carotid rete (Figure 16-40),uses
tained running and in race horses, greyhounds, and sled cool venous blood returning from respiratory passages to
dogs after racing. remove heat from hot arterial blood traveling toward the
The rise in Tb(and in the error signal as Tb rises above brain. In these animals, most of the blood to the brain flows
Ts,,) is kept small by the high sensitivity of the feedback through the external carotid artery. At the base of the skull,
control of heat-dissipating mechanisms. For example, a the carotid anastomoses into hundreds of small arteries that
small increase in T,,above the set-point temperature pro- form a vascular rete, the vessels of which rejoin just before
duces a strong and steep increase in the rate of sweating passage into the brain (see Figure 16-40). These arteries
(Figure 16-39).The effectiveness of heat loss is affected by pass through a large sinus of venous blood, the cavernous
humidity of the surrounding air-the higher the humidity, sinus. This venous blood is significantly cooler than the ar-
the less effective the heat loss (as any person familiar with terial blood because it has come from the walls of the nasal
either desert or high-humidity environments can testify). passages, where it was cooled by respiratory air flow. Thus,
The heat-loss mechanisms are initiated by vigorous exercise the hot arterial blood flowing through the rete gives up
even before peripheral body temperature has undergone some of its heat to the cooler venous blood before it enters
any significant increase. For example, in human beings, an the skull. As a result, brain temperature may be from 2 to
increased rate of sweating begins within 2 seconds after on- 3 Celsius degrees lower than the core temperature of the
704 INTEGRATION OF PHYSIOLOGICAL SYSTEMS
...............................................................................
Figure 16-40 The sheep has a carotid rete for counter-
current cooling of carotid blood. The carotid rete, found
in sheep and some other mammals, is shown in red. A
,,
/
network of small arteries acts as a heat exchanger for
Circle of ,, blood supplying the brain. Cool venous blood returning
Bin ,Willis ,/
from the nasal cavity bathes the carotid rete contained
Cavernous ,
/ in the cavernous sinus, removing heat from arterial
sinus \
blood flowing t o the circle of Willis and then t o the
brain. [Adapted from Hayward and Baker, 1969.1
Carotid
artery
body. Although sustained running in hot surroundings in- rousing to scan the ice for approaching polar bears. Human
evitably places a heat load on these animals, the most seri- beings and many other mammals sleep for hours at a time.
ous and acute consequence of overheating-spastic brain Many of the big carnivores (e.g., lions and tigers) sleep for
function-is thereby prevented. This system of cooling is as long as 20 hours a day, especially after a meal.
most effective when the animal is breathing hard during
strenuous exercise. Torpor
The lower the Tb, the lower the basal metabolism and the
DORMANCY SPECIALIZED lower the rate of conversion of energy stores, such as fatty
METABOLIC STATES tissues, into body heat. Thus, it is generally advantageous
to allow body temperature to decrease during periods of
Dormancy is a general term for reduced body activities, in- nonfeeding and inactivity. Small endotherms, because of
cluding reduced metabolic rate. It often includes het- their high rates of metabolism, are subject to starvation
erothermy. Dormancy can be variously classified according during periods of inactivity when they are not feeding. Dur-
to its depth (in reference both to ability for arousal and to ing those periods, some animals enter a state of torpor, in
decrease in Tb)and its duration, and it includes sleep, tor- which temperature and metabolic rate subside. Then, be-
por, hibernation, winter sleep, and estivation. Sleep has fore the animal becomes active, its body temperature rises
been the most thoroughly investigated (probably because it as a result of a burst of metabolic activity, especially
is the only state of dormancy experienced by people). The through shivering or oxidation of brown-fat stores or both
remaining four categories are less well understood than (if a mammal). Daily torpor is practiced by many terrestrial
sleep; however, in homeotherms, all appear to be manifes- birds. The hummingbird is a classic example, allowing its
tations of physiologically related processes. body temperature to fall from a daytime level of about
40°C to a nighttime level as low as 13°C (in the rufous
Sleep hummingbird when a low ambient temperature permits).
Studied intensively in human beings and other mammals, Several species of small mammals also undergo torpor (e.g.,
sleep entails extensive adjustments in brain function. In shrews), but large mammals have too much thermal mass
mammals, slow wave sleep is associated with a drop in to cool down quickly for short periods of torpor.
both hypothalamic temperature sensitivity and body tem-
perature, as well as changes in respiratory and cardiovas- Hibernation and Winter Sleep
cular reflexes. During rapid-eye-movement (REM) sleep, A period of deep torpor, or winter dormancy, hibernation
hypothalamic temperature control is suspended. Although lasts for weeks or even several months in cold climates. It is
there may be a variety of triggers of sleep, in mammals entered into through slow wave sleep and is devoid of
there is evidence of sleep-inducing substances that build up rapid-eye-movement sleep. Hibernation is common in
during wakefulness, accumulating in extracellular fluids of mammals of the orders Rodentia, Insectivora, and Chi-
the central nervous system. The identity and mode of action roptera, which can store sufficient energy reserves to sur-
of these substances is under investigation. The time course vive the periods of nonfeeding. Many hibernators arouse
and extent of sleep varies greatly among animals. Seals rest- periodically (as often as once a week or as infrequently as
ing on pack ice sleep for only a few minutes at a time before every 4-6 weeks) to empty their bladders and defecate.
USING EN
.......................................
During hibernation, the hypothalamic thermostat is reset arousal requires less than 3 hours. The speed at which this
to as low as 20 Celsius degrees or more below normal. At midsized mammal arouses depends on rapid heating initi-
ambient temperatures between 5°C and 1S0C, many hi- ated by intensive oxidation of brown fat, accompanied by
bernators keep their temperatures as little as 1degree above shivering. This frequently leads to a large surge in meta-
ambient temperature. If the air temperature falls to dan- bolic rate, as evident in Figure 16-42.
gerously low levels, the animal increases its metabolic rate Although many small endotherms undergo a daily cy-
to maintain a constant low Tbor becomes aroused. cle of torpor, their high rates of metabolism preclude ex-
Thermoregulatory control is not suspended during tended periods of torpor in the form of hibernation be-
torpor and hibernation-it merely continues with a low- cause, even in the hibernating state, they would quickly
ered set point and reduced sensitivity (gain), as in slow consume stored energy reserves with little remaining for the
wave sleep. In the hibernating marmot, for example, ex- metabolically expensive process of arousal. All true hiber-
perimental cooling of the anterior hypothalamus with an nators are midsized mammals weighing at least several
electronically controlled, implanted probe increases meta- hundred grams and large enough to store sufficient reserves
bolic production of body heat. The increase in heat pro- for extended hibernation. There are no true hibernators
duction is proportional to the difference between the among large mammals. Bears, which were once thought to
set-point temperature and the actual hypothalamic temp- hibernate, in fact simply enter a "winter sleep" in which
erature. The set-point temperature drops about 2.5 Celsius body temperature drops only a few degrees, and they re-
degrees within a day or two as the animal enters a deeper main curled up in a protected microhabitat such as a cave
state of hibernation. or hollow log. With its large body mass and low rate of
As might be expected, body functions are greatly slowed heat loss, a bear can store sufficient energy reserves to enter
with the lowered body temperature characteristic of torpor winter sleep without dropping body temperature. Bears are
and hibernation. The effect of reduced body temperature on able to wake and become active quickly at any point dur-
the metabolic rate (expressed asvCo,) of golden-mantled ing the winter, making it dangerous to encounter a bear
ground squirrels is shown in Figure 16-41. In conjunction even if it is in winter sleep. Typically, however, bears stay in
with a decrease in metabolism, blood flow in hibernating winter sleep for long periods, retaining metabolic wastes in
mammals is typically reduced to about 10% of prehiberna- their bodies and even bearing their cubs. Winter sleep, with
tion values, although the head and brown-fat tissue receive its relatively high body temperature, does not offer the
a much higher blood flow than do other tissues. The cardiac same degree of energy savings as deep hibernation, but a
output decreases to only a small percentage of the normal fall in body temperature of even a few degrees saves energy.
rate. This retardation is accomplished by a drastic slowing Why are there no large hibernators? First, they have less
of the rate of heartbeat, with stroke volume remaining es- need to save fuel, because their normal basal metabolic
sentially unchanged. As a result of reduced respiratory ex- rates are low relative to their fuel stores owing to the al-
change, the blood of many hibernators becomes more acidic. lometry of metabolism and fuel storage. Second, because of
This acidosis may further lower enzyme activity because of the large mass and relatively low rate of metabolism, a pro-
the departure from the pH optimum of metabolic enzymes. longed metabolic effort would be required to raise body
The rate of arousal from hibernation is often much temperature from a low level near ambient temperature to
higher than the rate of entry into hibernation. Thus, in the normal body temperature. It has been calculated, for ex-
ground squirrel, the transition to the torpid state is com- ample, that a large bear would require at least 24-48 hours
pleted within 12 to 18 hours (Figure 16-42), whereas to warm up to 37°C from a hibernating temperature of
5°C. Warming of such a large mass would also be energet-
ically very expensive.
Estivation
The poorly defined term estivation, which has been called
"summer sleep," refers to a dormancy that some species of
both vertebrates and invertebrates enter in response to high
ambient temperatures or danger of dehydration or both.
Land snails such as Helix and Otala become dormant dur-
ing long periods of low humidity after sealing the entrance
to the shell by secreting a diaphragm-like operculum that
retards loss of water by evaporation. Many land crabs sim-
ilarly spend dry seasons in an inactive state at the bottom
Temperature ("C) of their burrows. Well known as estivators are African
lungfish, Protopterus. These air-breathingfish survive peri-
Figure 16-41 Both experimentally induced hypothermia and natural hi-
bernation reduce metabolism in the golden-mantled squirrel. Open sym-
ods of drought in which their ponds dry up by estivating in
bols, experimentally induced hypothermia; asterisk, unanesthetised an- the semidry bottom until the next rainy season floods the
imals awake and hibernating. [From Milsorn, 1992.1 area. The lungfish seals itself inside a "cocoon," in which a
Hours Weeks Hours
Figure 16-42 Metabolism increases briefly during an episode of arousal for body temperature is depressed. Metabolism decreases, allowing T,
from hibernation in a ground squirrel. The squirrel was kept in a chamber to drop to 1-3 Celsius degrees above T,throughout hibernation.Arousal
having a temperature of 4°C. The period of steady-state hibernation is occurs when the set-point temperature climbs to 38'C, and a strong
shaded in color, and the body temperature, Tb,is graphed in red. Me- surge of metabolic heat production raises Tb to the new set-point level.
tabolism is graphed in black. At onset of hibernation, the set point Abbreviation: RAMR, resting average metabolic rate. [From Swan, 1974.1
small tube leads from the fish's mouth to the exterior to al- is usually expressed in units of kilocalories per kilogram per
low ventilation of the lungs. Interestingly, chemical estiva- kilometer. This energy is taken as that expended above and
tion-inducing factors in the plasma of estivating lungfish beyond that which is expended under basal conditions of
produce a torporlike state when injected into mammals. rest. Measurements of 0, consumption and CO, produc-
Some small mammals, such as the Columbian ground tion associated with locomotion are generally made while
squirrel, spend the hot late summer inactive in their bur- the animal is running on a treadmill, swimming in a flow
rows, with their core temperatures approaching the ambi- tank, or flying in a wind tunnel. The measured rate of gas
ent temperature. This state is probably similar physiologi- exchange is then translated into rate of energy conversion.
cally to hibernation, but it differs in seasonal timing. Relations between the net work done in the locomotion
of an animal and the gross energy conversion powering the
underlying muscle activity are complicated by several fac-
tors, not all of which are sufficiently well understood to be
discussed here. Nonetheless, we know that a significant per-
centage of muscular effort during locomotion does not con-
tribute directly to the production of forward motion. Some
muscle contraction holds limb joints in their proper articu-
lating positions. Another large percentage of muscle work
ENERGETICS OF LOCOMOTION is performed in an elingating muscle to counteract gravity,
Early in this chapter, we considered the basal rate of me- to absorb shocks, and to finely tune the movements of
tabolism characteristicof the resting animal. Additional en- limbs during contraction of antagonists. The comparative
ergy over and above the basal rate is expended when the energetics of animal locomotion are further complicated by
animal is active-that is, producing movement with its the well established inverse relation between the force pro-
muscles. The most readily quantified type of muscle activ- duced by a contracting muscle and its rate of shortening
ity in most animals is simple locomotion. Because it is re- (i.e., muscle length or sarcomere length per second; see
quired for finding food and mates and for escaping preda- Figure 10-13).The higher the rate of cross-bridge cycling,
tors, locomotion is also one of the more important types of the higher the metabolic cost of shortening the muscle by
routine activity. We now turn to an examination of the a given distance. Small animals show higher rates of limb
metabolic cost of animal locomotion. stride, tail beat, or wing motion. Thus, small animals em-
ploy higher rates of muscle shortening (and hence cross-
Animal Size, Velocity, and Cost of Locomotion bridge cycling) to achieve a given velocity of locomotion
The metabolic cost of locomotion is the amount of energy than do larger animals. For that.reason, they must convert
required to move a unit mass of animal a unit distance and correspondingly larger amounts of metabolic energy to
USING ENERGY: MEETING ENVIRONMENTAL CHALLENGES 707
...............................................................................
produce a given amount of force per unit cross section of terrestrial animals moving at low and moderate speeds
contractile tissue in moving their limbs. through air, where drag is negligible. More likely, the lower
Several generalizations can be made in relating the energy efficiency is related to the lower efficiency of rapidly
overall energy cost of locomotion to the size and velocity of contracting muscle.
an animal. Locomotion is a metabolically expensive pro- The relation between velocity and the cost of locomo-
cess. The rate of oxygen consumption in excess of the basal tion is complex. As velocity of running increases in
metabolic rate increases linearly with the velocity quadripedal mammals, for example, the metabolic cost of
(Figure 16-43A). It is noteworthy, however, that the in- traveling a given distance initially decreases (Figure 16-44).
crease in energy utilization per unit weight for a given in- This is because nonlocomotory expenses account for a pro-
crement in speed is less for larger animals than for smaller gressively smaller fraction of the total energy expended.
ones. This can be seen in the different slopes of the plots in However, as velocity continues to increase, animals that
Figure 16-43A. When the cost of locomotion is plotted as swim, fly, or run all begin to experience an increase in the
energy utilization per gram of tissue per kilometer against cost of locomotion as they generate near maximal locomo-
body mass, it is again apparent that larger animals expend tory velocities. Figure 16-45 illustrates this phenomenon in
less energy to moue a given mass a given distance cephalopods (e.g., squids and nautilus), for which a typical
(Figure 16-43B).The lower energy efficiency of small ani- U-shaped curve describes the cost of locomotion at various
mals during locomotion may, to a limited degree, be due to locomotory velocities. The cost of locomotion initially de-
the greater drag that they experience (to be discussed creases sharply as velocity increases but then begins to rise
shortly), but this explanation certainly does not suffice for at higher velocities.
d
2-
R u n n ~ n speed
g (km/ h)
Weight
708 INTEGRATION OF PHYSIOLOGICAL SYSTEMS
..............................................................................
I Wh~temouse (21 g)
2 Kangaroo rat (41 g)
3 Kangaroo rat (I00 g)
4 White rat (384 g)
5 Ground squlrrel (236 g)
6 Dog (2 6 kg)
7 Dog (18 kg)
Speed (m s-').
Figure 16-46 Hopping, bipedal animals such as wallabys and kangaroos
can increase velocity with no increase in oxygen consumption. Similar-
2 4 6 8 10 sized quadrapeds and wallabys initially show a linear increase in oxygen
consumption as velocity increases. As wallabys switch t o bipedal loco-
Running velocity (km . h-')
motion, however, their rate of oxygen consumption does not increase
Figure 16-44 The energetic cost of transporting a unit of body mass by with further increases in velocity. That blood lactate similarly stays level as
running decreases as body size increases in mammals. The cost of run- velocity increases indicates that the higher speeds are not achieved by
ning 1 km drops and levels off with increasingvelocity. Dashed propor- an increase in anaerobic metabolism. [Adapted from Baudinette, 1991.1
tions are extrapolated. [From Taylor et al., 1970.1
Swimming
Animals that swim in water need to support little or none
of their own weight. Many have flotation bladders or large
amounts of body fat that enable them to suspend them-
selves at a given depth with little expenditure of energy.
However, although the high dellsity of water allows them
to be neutrally buoyant, it also produces high drag. This
hindrance to objects moving through a fluid has led to a
Time convergence of body forms among marine mammals and
fishes. The streamlined, fusiform body shape is wonderfully
developed in most sharks, teleost fishes, and dolphins. The
reasons are evident enough on intuitive grounds, but they
can be understood more clearly in relation to flow pattern.
The ease with which an object moves through water de-
pends in part on the flow pattern of the water. The fluid at
the immediate surface of the object moves at the same ve-
locity as the object, whereas the fluid at a great distance is
undisturbed. If the transition in fluid velocity is smoothly
continuous as the fluid progresses away from the object's
Time surface, then laminar flow (Chapter 12) occurs at the
boundary layer-the layer of unstirred fluid immediately
Figure 16-47 Body mass affects both rate of energy expenditure and ac-
celeration during locomotion. (A) Rate at which energy is used per unit
adjacent to the object's surface (Figure 16-48A). In con-
mass during onset and maintenance of locomotion (shaded region) in a trast, turbulent flow results when there are sharp gradients
large and a small animal of similar type. (6)Velocity of a small and a large and inconsistencies in the velocity of flow. Because of con-
animal during acceleration and deceleration atthe beginning and end of servation of energy, pressure and velocity are reciprocally
a period of locomotion (shaded region). related in a given fluid system, and the higher the velocity
of fluid at a given site, the lower the pressure. Thus,
strongly differing flow rates around an object cause eddy
moves. The drag produced in a given medium depends on currents owing to secondary flow patterns set up between
the velocity, surface area, and shape of an object. For an ob- regions of high and regions of low pressure. Moreover, the
ject of a given shape, drag is proportional to the surface higher the viscosity of the medium or the higher the relative
area. Because larger animals have lower surface-to-mass motion between the object and the surrounding fluid, the
ratios, they experience less fluid drag per unit mass than greater the shear forces produced and, hence, the greater
do smaller animals, for whom overcoming drag is energet- the tendency toward turbulence. Because its production
ically more costly. Once it is under way, a larger animal dissipates energy as heat, turbulence retards the efficient
expends less energy per unit mass to propel itself at a given conversion of metabolic energy into propulsive movement.
velocity than does a smaller animal of similar type (see Long, streamlined shapes promote laminar flow with
Figure 16-47A).Drag is also proportional to the square of minimal eddy current formation. Fishes and marine mam-
an animal's velocity, meaning that the energy required to mals such as seals, porpoises, and whales are admirably
overcome drag and propel the animal at faster speeds in- streamlined, exhibiting nearly turbulence-free passage
creases with velocity. through water even at high speeds. Flying birds are similarly
h
These effects are far more pronounced in water than in streamlined in flight. An additional factor reducing turbu-
air because water, having the higher viscosity and density, lence in these animals is the compliance (deformability) of
produces far more drag on a moving object than air does. the body surface. A high body compliance damps small per-
Drag is of major importance in swimming and flying be- turbations in the pressure of the water flowing over the
cause of the high viscosity of water faced by swimmers and body surface and thereby lessens the local variations in wa-
the high velocity experienced by flyers. Drag is of little im- ter pressure that give rise to energy-dissipating turbulence.
portance in running, because the velocities attained in run- The speed of a swimming animal is proportional to the
ning are low, as is the viscosity of air. These relations are power-to-drag (thrust-to-drag) ratio. Power developed by
quantified in the Reynolds number (see Spotlight 16-2). contracting muscle is directly proportional to muscle mass,
and, if we assume that muscle mass increases in proportion
Aquatic, Aerial, and Terrestrial Locomotion to total body mass, power (thrust) rises in proportion to
Animals have evolved myriad ways of moving in water, on body mass. On the other hand, for a large swimming ani-
land, and in air. Despite this diversity, each mode of loco- mal, total drag increases but drag per unit mass decreases,
Reynolds number
for a given velocity, as body mass increases. This is because, to the relatively low drag forces developed, birds can
if the shape remains constant, the surface and cross-sec- achieve much higher speeds than those of fishes. The
tional areas (which determine the drag) increase as a func- production of propulsive force, which drives the bird
tion of some linear dimension squared, whereas body mass forward, and lift, which keeps it aloft, is accomplished
(which determines the power available) increases as a func- simultaneously during the downstroke of the bird wing
tion of that linear dimension cubed. Thus, a large aquatic (Figure 16-49). The wing is driven downward and forward
animal can develop power out of proportion to its drag with an angle of attack that pushes the air both downward
forces; therefore, it is able to attain higher swimming ve- and backward, creating an upward and forward thrust.
locities than thosi of a smaller animal of the same shape. The components of lift and forward propulsion overcome
Large fishes and mammals swim faster than their smaller the bird's weight and drag, respectively.
counterparts. Because of the high drag forces developed in The body shapes of fishes and birds demonstrate the
water and because drag increases as the square of velocity, great differences that exist between the physical properties
aquatic animals can reach the speeds of a bird in flight only of water and those of air, as well as the biological diver-
if they are much larger and more powerful than the bird. gence that results from adaptation to these two dissimilar
media. When a bird is gliding, its elongated, extended
Flying wings that take the form of an airfoil produce excellent lift;
Unlike water, air offers little buoyant support, so all flyers but, if the bird were in water, they would obviously gener-
must overcome gravity by utilizing principles of aerody- ate far too much drag. Thus, the wings of penguins are
namic lift. Although the effects of drag increase with speed, modified to serve as short paddles and are folded against
there is still less need for streamlining among birds than the body while the birds are coasting under water. Because
among fishes because of the low density of air. Thus, thanks drag forces are much higher in water than in air, only
ciency is lost because the antigravity extensor muscles that
contract to propel the CM upward and forward must also
break the fall of the CM that occurs before the next stride.
To control the fall, the extensor muscles must expend en-
ergy to resist lengthening as they slow the rate of descent of
the body in preparation for the next cycle. This technically
unproductive use of muscle energy to counteract the pull of
gravity is said to produce "negative work." You may be fa-
miliar with such negative work carried out by your legs' ex-
tensor muscles while hiking down a steep trail.
In short, running or walking is less efficient than flying,
N Angle of attack swimming, or bicycling because the muscles must be used
for deceleration (negative work) as well as acceleration
(positive work). One reason that a person riding a bicycle
DL Local drag
is so efficient (and thus why people can cycle many times
faster and farther than they can run) is that the CM does
T, Local thrust
not rise and fall, which means that more muscular energy
can be transferred into forward velocity.
Elastic energy storage in elastic elements of the limbs
appears to be especially important in running and hopping
animals. Consider the bounding of a kangaroo. The greater
the height achieved during the hop, the greater is the speed
of descent and, when the legs strike the ground, the more
energy is transferred to the elastic elements in the limbs and
Figure 16-49 The downstroke of a bird wing develops forces in several the greater the force of elastic recoil ofthe limbs when they
directions. (Top) the wing in stage 3 of the flight cycle shown at the bot-
tom. Red arrows illustrate forces relating to the wingbeat; black arrow
subsequently extend at the beginning of the next hop. Not
illustrates force relating to the body. lnduced drag equals the drag many but the concept of
produced as a consequence of lift production. Induced thrust is corn~le- elastic energy storage is important when considering
mentary to induced drag. [From Nachtigall, 1977.1 changes in gait (e.g., walking to trotting to galloping in a
horse). By changing gait at appropriate speeds, land ani-
mals enhance their locomotory efficiency and avoid poten-
medium-sized to large animals can coast in water. In tially injurious forces on legs. For example, consider a pony
contrast, only very small flying animals, smaller than a made to trot on a treadmill at a speed at which it would
dragonfly, are unable to coast (glide) in air. Small insects normally gallop, or to gallop when it would normally trot,
such as flies and mosquitoes must continually beat their or to trot when it would normally walk. In all cases, it ex-
wings to maintain headway, because they have very little pends more energy than if allowed to change its gait natu-
momentum. rally. Optimum gaits result from the relative amounts of en-
ergy stored in the elastic elements of the body, such as
Running tendons, when performing the different gaits. For instance,
When swimming, flying, and running are compared with little energy is stored when an animal walks; somewhat
respect to the energy cost of moving a given body mass more is stored when it trots. When the animal is galloping,
a given distance (Figure 16-50), it is apparent that terres- its entire trunk is involved in elastic storage. At least half the
trial locomotion (i.e., running) is the most costly, whereas negative work done in absorbing kinetic energy during the
swimming is the least expensive. A swimming fish expends stretching of an active muscle appears as heat; the remain-
less energy in locomotion than a bird does in flying through der is stored in stretched elastic elements such as muscle
the air because, as already noted, the fish is close to neu- cross-bridges, sarcoplasmic reticulum, and Z-lines of mus-
tral buoyancy, whereas a bird must expend energy to stay cle cells and tendons. Only the elastically stored energy is
aloft. But why is running less efficient than either flying or available for recovery on the rebound, and only from 60%
swimming? to 80% of that is recovered on its release. The energy con-
Running differs from swimming and flying in the way verted into heat is not available for conversion into me-
limb muscles are used, and this difference accounts for the chanical work in living tissue.
low work efficiency of running. When a biped or qua-
druped animal runs, its center of mass (CM)rises and falls Locomotory energetics of ectotherms versus endothewns
cyclically with the gait. The rise in the CM occurs when the You might think that, on simple energetic grounds, terres-
foot and leg extensors push the body up and forward, and trial endotherms and ectotherms of the same size will ex-
the fall occurs as gravity inexorably tugs at the body, bring- pend the same net metabolic energy to run at a given ve-
ing it back to earth between locomotory extensions. Effi- locity. This reasonable assumption is almost, but not
712 INTEGRATION OF PHYSIOLOGICAL SYSTEMS
...............................................................................
Crawlers, F l ~ e r s Sw~mmers
runners
Insects V
V- Mosqu~to
F~shes
Reptiles o
B~rds 0 A V
Mammals o A V
Mach~nes
Dtr~g~ble
Bicycl~st
W Tractor traller/truck
I Fre~qhtsteamer
Figure 16-50 Cost of transport is more closely related to the kind calories per gram per kilometer for animals as well as machines. [From
of locomotion than it is t o the kind of organism. Cost is given in kilo- Tucker, 1975.1
entirely, correct. When 0, consumption is plotted against dotherm of a given size can achieve a higher rate of activity
running velocity for a lizard and a mammal of similar size, while undergoing aerobic metabolism than a similar-sized
the aerobic parts of the plots for both exhibit rather similar ectotherm can.
slopes. Thus, when movement begins, a similar increment The locomotor speed at which the maximum rate of
in metabolic energy expenditure is required for a similar in- aerobic respiration is reached is termed the maximum aer-
cremental increase in velocity for both the mammal and the obic velocity (MAV).As an animal exceeds its maximum
lizard. The difference between the two animals lies in the aerobic velocity, the additional activity is supported entirely
lower y-intercept of the lizard's plot relative to its standard by anaerobic metabolism, which leads to glycolytic pro-
metabolic rate while at rest. The reason for the differences duction of lactic acid. As lactic acid production progresses,
between the resting metabolic rates and the y-intercepts is an oxygen debt develops (see Metabolic scope, earlier in
not completely certain, but these differences may represent this chapter).Anaerobic metabolism is also associated with
the "postural cost" of locomotion-this cost being higher muscle fatigue (due to progressive depletion of chemical en-
for a mammal than for a lizard. ergy stores) and metabolic acidosis, which if extreme can
As noted earlier, the rate of 0, consumption rises lin- disrupt tissue metabolism. Because of these two conse-
early with increasing velocity of locomotion. This relation quences, anaerobic metabolism is unsuitable for sustained
is true for ectotherms as well as for endotherms. An en- activity. Thus, only locomotion below the maximum aer-
dotherm of a given mass typically has a basal metabolic obic speed can be sustained by animals of either group. Be-
rate about 6 to 10 times that of an ectotherm of similar cause endotherms are capable of far higher rates of aerobic
mass. In both groups, a similar relation exists between the metabolism than are ectotherms, they are generally capable
basal rate and the maximum metabolic rate that can be of higher rates of sustained locomotor activity.
achieved with intense exercise. That is, the factorial scope Clearly, the implications for ectothermy and en-
for locomotion exhibited by both groups is about the same. dothermy are not limited to mechanisms of temperature
Thus, in response to intense exercise, endotherms can control but are also of great importance to the kinds of ac-
achieve a maximum rate of 0, consumption of as much as tivity that an animal can undertake. The metabolic differ-
tenfold that of ectotherms of similar size, and so an en- ences between ectotherms and endotherms determine, for
U S I N G ENERGY: M E E T I N G E N V I R O N M E N T A L C H A L L E N G E S 713
...............................................................................
example, how far and how fast they can travel. This is not Circadian Rhythms
to say that ectotherms cannot achieve rates of activity and Biological rhythms lasting from milliseconds (at the cellu-
speeds of locomotion as high as those of endotherms. How- lar level) to years (at the whole-animal level) have been
ever, because locomotor activity in excess of the maximum identified in a wide variety of animals. Most rhythms (and
aerobic velocity relies on prodigious rates of anaerobic me- certainly many of the most prominent and well studied
tabolism, high rates of locomotion can be sustained by ec- rhythms) relate to daily cycles, which are called circadian
totherms for only brief periods. This limitation can be ob- rhythms. A true circadian rhythm, which is endogenously
served in ectothermic vertebrates such as some species of generated, can be distinguished from a physiological or
lizards and frogs, in which a rapid burst of activity, seldom other variable that just happens to track daily changes in
lasting more than a few seconds, takes the animal quickly environment by the use of four different criteria.
to a new resting or hiding place when disturbed. In some First, a circadian rhythm shows persistence, remaining
fishes, more than 50% of body mass is glycolytic white for at least several days or weeks in an animal that has been
muscle fibers specialized for very short bursts of locomotor removed from the natural environment and placed in a lab-
activity. The disadvantage that ectotherms have in sustain- oratory setting with constant environmental conditions
ing high rates of activity is offset by their more modest en- (constant temperature, constant light or constant dark,
ergy requirements, which enable them to spend more time etc.). A true circadian rhythm will persist in the animal,
hiding and less time looking for a meal. manifest most often as a continuation of that animal's nor-
mal daily cycles, the innate period being near 24 hours. Al-
though anv of a variety of physiological or behavioral char-
Why can fleas leap hundreds of times their own acteristics can be measured, one of the most frequently
body length, whereas most medium-sizedmam- measured is general activity. Figure 16-51 shows a typical
mals can jump only a few body lengths, and apparatus for measuring the locomotor activity of a rodent.
large mammals do not even attempt to leap? When the animal runs ~n the exercise ivheel, the ~ctivityis
Consider both the physical surroundings of the recorded, either directly on a chart recorder or on a
animals and their own structural makeup.
-
BODY RHYTHMS AND ENERGETICS
Most animals strive to maintain some sort of constancy in
-
their interior milieu. Although variations in body tempera-
ture, metabolic rate, intracellular pH, and body energy con-
tent, for example, may of necessity fluctuate widely in re-
sponse to environmental constraints and demands, most
animals have a preferred range for these and other physio-
logical variables. Despite the evolution of mechanisms that
Iogg.11 17-\ Activity record
-
cles falls by a half degree or so during the early morning profile
10
hours (about 3:OO-5:00 A.M.), only to rise again at about
normal waking time. In fact, virtually all animals and 5
plants show some type of rhythmic variation in metabolism
or some other physiological variable. Biological rhythms Light-dark cycle 1
are so innate to animals that even individual cells in cell cul-
Figure 16-51 Circadian rhythms can b e recorded by monitoring spon-
ture will show rhythms in rate of cell division. The cell does taneous activity. In this example, a rodent runs in an exercise wheel, which
not have to be a particularly complex cell-a daily rhythm activates a switch closing an electric circuit every time it turns. The result-
occurs, for example, in prokaryotic cells such as nitrogen- ing electrical record of activity can b e written direaly on a chart recorder
fixing cyanobacteria. or, more conveniently, logged in a computer for later analysis.
computer. The apparatus can be modified by replacement oxygen levels, and so forth, can disrupt the normal circa-
of the exercise wheel with some other form of activity- dian rhythm. Often, there is a threshold level of tempera-
measuring device to record activity in birds, fishes, or vir- ture below which the rhythm is finally disrupted. The ef-
tually any other animal. Figure 16-52 shows activity levels fects of light are more graded. In the mosquito, forexample,
in a blinded house sparrow. For the first two weeks of the a circadian activity rhythm is evident when the light phase
record, the sparrow was in a light-dark cycle. Even though of the light-dark cycle consists only of low light, but this
the bird was unable to see, its circadian rhythm nonetheless rhythm gradually diminishes as light intensity increases.
persisted, with a free-running period several minutes longer A final characteristic feature of circadian rhythms is
than 24 hours. that they can be entrained. If an animal is placed in total
The second characteristic feature of circadian rhythms darkness, for example, the length of its circadian rhythm re-
is that they tend to be largely body-temperature indepen- mains close to 24 hours but is generally slightly shorter or
dent. We have seen that metabolism and physiological longer, resulting in a progressive "creep" in activity cycles
processes that derive from and contribute to metabolism that is quite obvious in a long-term record of an animal's
have a temperature quotient, Q,,, - of between 2 and 3. Yet,
A
activity (see Figure 16-52). However, if the animal in dark-
a rise in temperature typically causes very little or no in- ness is then given a new lighting regime with a periodicity
crease in the cycling of the circadian rhythm; in some ani- of slightly longer or shorter than 24 hours, the animal's ac-
mals, an increase in body temperature may actually slow tivity will be entrained to the new lighting regime. Reen-
down the circadian rhythm. trainment is not instantaneous but takes place in a series of
Circadian rhythms are also characterized by the fact transients that manifest the inability of the internal clock to
that they can be made conditionally arrhythmic-that is, a be shifted more than a certain amount each cycle. Through
certain set of environmental temperatures, lighting regimes, the use of new light cues, activity cycles can be advanced or
delayed to the point of being in a completely opposite phase
to that of the original circadian rhythm. In the experiment
One day's- with the blinded house sparrow (see Figure 16-52), the
activity feathers were removed from the top of its head. This al-
lowed light to penetrate through the skull to the brain,
Feathers removed parts of which are light sensitive, and the sparrow's activity
from top of skull - was then entrained to the light-dark cycle. As the feathers
grew in, the entrainment began to be lost and the circadian
rhythm lengthened, but removal of the feathers for a sec-
ond time returned the entrained activity rhythm. In a final
experiment, injection of dye under the scalp on top of the
skull blocked light penetration to the brain, and the circa-
dian activity rhythm began to drift once again.
Light is the most commonly effective zeitgeber, or en-
Newly grown vironmental entrainment factor. However, environmental
feathers removed
from top of skull - temperature, food availability, and interactions with other
animals of the same or different species also may be zeit-
gebers that exert an effect on metabolism, activity, and
Dye injected - other basic facets of animal life.
under skin
covering skull
rhythms correlate with the 29.5 day lunar cycle and affect
reproduction in many animals. Circannual cycles correlate 2
with the 365 day earth year and are most evident in the 2." 0
often strong, seasonal cycles that affect everything from fur I
color to hibernation to migration of many animals. Note
that all of the circ- rhythms are endogenous rhythms-
they persist if external cues are removed, and they are en- 5
trainable.
w
feeding feeding
plete bed rest. In reality, circadian rhythms in body tem- 18 -
perature are often imposed on activity rhythms with a sim-
16
ilar time component, amplifying the daily range of body -
temperature. 14 -
Ultradian rhythms in body temperature are best exem-
12 -
plified by the hibernators, which, as noted earlier, lower
their core temperature by 20-35 Celsius degrees for peri-
ods of weeks or months, punctuated by brief periods of
10 -
8-
t 1
arousal. These rhythms can persist for at least four years in
hibernating golden-mantled ground squirrels (Citellus) iso-
lated at birth from any light or temperature zeitgebers. In
hibernating bats, circadian rhythms in body temperature
4
6-
I
- Night
Physical quantity Name of unit Symbol for unit Multiplier Prefix Symbol
Derived SI units
- -
-
IJ
Abscissa x
Exponential equations:
In biological systems there is often an exponential relationship between values, described by the equation:
y = b.ax
The logarithmic form of this equation is:
logy = logb + xloga
Using semi-log graph paper (linear abscissa) log y can be plotted against x, giving a straight line to determine the values of
slope a and intercept b.
X X
(Examplesof the use of straight line and exponential equations may be found throughout the book, especially in chapters
3 and 16.)
Use of logarithm terms:
, In the equation y = lox,x is the logarithm of y. That is, x is the power to which 10 must be raised in order to yield y. For
example, the logarithm of 10 is 1,and the logarithm of 100 is 2. The equation
atmospheres bars
dynes per square centlmeter
grams per square centlmeter
torr (= mmHg; PC)
pounds per square ~nch
pascals
grams daltons
grams
ounces (avdp)
pounds (avdp)
inches angstroms
centimeters
feet
meters
joules calories
ergs
foot-pounds
watt-hours
watt-seconds
APPENDIXES 727
...............................................................................
Units and conversion factors (Continued)
meters angstroms
micrometers Cum)
centimeters
feet
inches
kilometers
miles (statute)
millimeters
yards
newtons dynes
Temperature conversions
.. ~
Power p = w/t
w = work Kinetic energy E = 1/2mv2
t = time v = velocity of mass
Chemical definitions
1 rnol = the mass in grams of a substance equal to its molecular or
atomic weight: this mass contains Avogadrok number (N,)
of molecules or atoms
Molar volume = the volume occupied by a mole of gas at
standard temperature and pressure
(25"C, 1 atm) = 22.414 L
1 molal solution = 1 mol per 1000 g of solvent
1 molar solution= 1 mol of solute in 1 L of solution
1 equivalent = 1 mot of 1 unit charge
1 einstein = Irnol of photons
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Bruns, D., and R. Jahn. 1995. Real-time measurement of trans- Denton, E. J. 1961. The buoyancy of fish and cephalopods. Prog.
mitter release from single synaptic vesicles. Nature Biophys. 11:178 -234.
377:62-65. Diamond, J., and K. Hammond. 1992. The matches, achieved by
Biilbring, E. 1959. Lectures on the Scientific Basis of Medicine. natural selection, between biological capacities and their nat-
Vol. 7. London: Athlone. ural loads. Experientia 48:551-557.
Biilbring, E., and H. Kuriyama. 1963. Effects of changes in ionic Diamond, J. M., and J. McD. Tormey. 1966. Studies on thc struc-
environment on the action of acetylcholine and adrenaline on tural basis of water transport across epithelial membranes.
smooth muscle cells of guinea pig. J. Physiol. 166:59-74. Federation Proc. 25:1458 - 1463.
Bullock, T. H., and F. P. J. Diecke. 1956. Properties of an infrared Douglas, W. W. 1974. Mechanism of release of neurohypophyseal
receptor. J. Physiol. 134:47-87. hormones: stimulus-secretion coupling. In R.O. Greep, ed.,
Bullock, T. H., and G. A. Horridge. 1965. Structure and Function Handbook of Physiology. Section 7. Endocrinology (Vol. 4,
in the Nervous Systems of Invertebrates. New York: Part 1, Pituitary Gland). Washington, D.C.: American Physi-
W. H. Freeman and Company. ological Society.
Burggren, W., and K. Johansen.1982. Ventricular hemodynamics Douglas, W. W., J. Nagasawa, and R. Schulz. 1971. Electron
in the monitor lizard, Varanus exanthematicus: pulmonary microscopic studies on the mechanism of secretion of
and systemic pressure separation. J. Exp. Biol. 96:343-354. posterior pituitary hormones and significance of micro-
Camhi, J. M. 1984. Neuroethology. Sunderland, Mass.: Sinauer vesicles ("synaptic vesicles"): evidence of secretion by
Associates, Inc. exocytosis and formation of microvesicles as a by-product
Cannon, W. 1929. Organization for physiological homeostatics. of this process. In H. Heller and K. Lederis, eds., Sub-
Physiol. Rev. 9:399-431. cellular Organization and Function in Endocrine Tissues.
Capecchi, M. R. 1994. Targeted gene replacement. Scientific Mem. Soc. Endocrinol. No. 19. New York: Cambridge
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Carey, F. G. 1973. Fishes with warm bodies. ScientificAmerican Dudel, J., and S. W. Kuffler. 1961. Presynaptic inhibition at the
228:36-44. crayfish neuromuscular junction. J. Physiol. 155543-562.
Carey, F. G., and J. M. Teal. 1966. Heat conservation in tuna fish Eakin, R. 1965. Evolution of photoreceptors. Cold Spring Harbor
muscle. Proc. Nut. Acad. Sci. USA 56:1464-1469. Symp. Quant. Biol. 30:363-370.
Catania, K. C., and J. H. Kaas. 1996. The unusual nose and brain Ebashi, S. K. Maruyama, and M. Endo, eds. 1980. Muscle Con-
of the star-nosed mole. BioScience 46(8):578-586. traction: Its Regulatory Mechanisms. New York: Springer-
Chen, J-N., and M. Fishman. 1996. Genetic dissection of heart de- Verlag; pp. 10-62.
velopment. In W. Burggren and B. Keller, eds., Development Ebashi, S., M. Endo, and I. Ohtsuki. 1969. Control of muscle con-
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York: Cambridge University Press. Eccles, J. C. 1969. Historical introduction to central cholinergic
Chess, A., L. Buck, et al. 1992. Molecular biology of smell: - transmission and its behavioral aspects. Federation Proc.
-
alimentary canal A hollow, tubular cavity extending through ani- amphoteric Having opposite characteristics; behaving as either an
mals open at both ends; for ingestion, digestion and secretion acid or a base.
of food materials amylase Carbohydrases that hydrolyze all but the terminal glyco-
alkali earth metals A group of grayish-white, malleable metals eas- sidic bonds within starch and glycogen, producing disaccha-
ily oxidized in air, comprising Be, Mg, Ca, Sr, Ba, and Ra. rides and oligosaccharides.
alkaline tide A period of increased body and urinary alkalinity as- anabolism Synthesis by living cells of complex substances from
sociated with excessive gastric HCI secretion during digestion. simple substances.
alkaloids A large group of organic nitrogenous bases found in anaerobic Oxygen-free.
plant tissues, many of which are pharmaceutically active (e.g., anaerobic metabolism Metabolism not utilizing molecular
codeine, morphine). oxygen.
alkalosis Excessive body alkalinity. anastomose To interconnect.
GLOSSARY G-3
......................................
anatomical dead space The nonrespiratory conducting pathways found primarily in muscle; acts as a storage form of high-en-
in the lung. ergy phosphate for the rapid phosphorylation of ADP to ATP.
androgens Hormones having masculinizing activity. arteriole A tiny branch of an artery; in particular, one nearest a
aneurism Localized dilation of an artery wall. capillary.
angiotensin A protein in the blood, converted from angiotensino- arteriosclerosis A class of diseases marked by an increase in thick-
gen by the action of renin; it first exists as a decapeptide (an- ness and a reduction in elasticity of the arterial walls.
giotensin I) that is acted upon by a peptidase, which cleaves it association cortex Areas of the cerebral cortex that neither directly
into an octapeptide (angiotensin II), a potent vasopressor and receives sensory information nor directly contributes to mo-
stimulator of aldosterone secretion. tor output; instead neurons of association cortex typically re-
angiotensin II See angiotensin. ceive input from many sensory modalities and are broadly
anion Negatively charged ion; attracted to the anode or positive connected to other areas in the cortex and other brain centers.
pole. asynchronous muscle A type of flight muscle found in the thorax
anode Positive electrode or pole to which negatively charged ions of some insects; contracts at a frequency that bears no one-to-
are attracted. one relation to motor impulses. See also fibrillar muscle.
anoxemia A lack of oxygen in the blood. ATP See adenosine triphosphate.
anoxia A lack of oxygen. ATPase (adenosine triphosphatase) A class of enzymes that cat-
alyze the hydrolysis of ATP.
antagonist remuscle A muscle acting in opposition to the move-
ATPS Ambient temperature and pressure, saturated with water
ment of another muscle.
vapor; referring to gas volume measurements.
antagonists Agents that inhibit, block, or counteract an effect;
atria A chamber that gives entrance to another structure or organ;
for example, antagonists of synaptic transmitters typically
usually used alone to refer to an atrium of the heart.
block the postsynaptic receptor molecules that bind the
neurotransmitter. atrial natriuretic peptide (ANP) One of a family of peptide hor-
mones, cleaved from a single precursor peptide and produced
antenna1 gland Crustacean osmoregulatory organ.
in the cardiac atria, the physiological effects of which include
antibody An immunoglobulin, a four-chain protein molecule of a
increased urine output, increased sodium excretion, and a
specific amino acid sequence; an antibody will interact only
receptor-mediated vasodilation, the net result of whlch is
with the antigen that brought about its production or one very
lowered blood pressure.
similar to it.
atrioventricular node Specialized conduction tissue in the heart,
antidiuretic hormone (ADH,vasopressin) A hormone made in the which, along with Purkinje tissue, forms a bridge for electri-
hypothalamus and liberated from storage in the neurohy- cal conduction of the impulse from atria to ventricles.
pophysis; acts on the epithelium of the renal collecting duct by
auditory cortex Regions of the cerebral cortex that are associated
stimulating osmotic reabsorption of water, thereby producing
with hearing.
a more concentrated urine; also acts as a vasopressor.
auditory ossicle One of the bones of the middle ear (the malleus,
antigen A substance capable of bringing about the production of
the incus, and the stapes), connecting the tympatic membrane
antibodies and of then reacting with them specifically.
and the oval window.
antimycin An antibiotic that is isolated from a Streptomyces
autocrine A hormonal pathway characterized by the produc-
strain; acts to block electron transport from cytochrome 6 to
tion of a biologically active substance by a cell; the substance
cytochrome c in the electron-transport chain.
then binds to receptors on that same cell to initiate a cellular
antiports Membrane transport proteins that transfer two solutes, response.
each in the opposite direction.
autoinhibition Self-inhibition.
anus The opening of the alimentary canal through which feces are
autonomic nervous system The efferent nerves that control invol-
expelled.
untary visceral functions; classically subdivided into the sym-
aorta The main artery leaving the heart. pathetic and parasympathetic sections.
aortic body A nodule on the aortic arch containing chemorecep- autoradiography The process of making a photographic record of
tors that sense the chemical composition of the blood. the internal structures of a tissue by utilizing the radiation
apical Pertaining to the apex; opposite the base. emitted from incorporated radioactive material.
s apnea The suspension or absence of breathing. autorhythmicity The generation of rhythmic activity without ex-
apoenzyrneThe protein portion of an enzyme; the apoenzyme and trinsic control.
coenzyme form the functioning holoenzyme. autotrophic Pertaining to the ability to synthesize food from inor-
apolysis Release; loosening from. ganic substances by utilizing the energy of the sun or of inor-
aporepressor A repressor gene product that, in combination ganic compounds.
with a corepressor, reduces the activity of particular structural Avogadro's law Equal volumes of different gases at the same tem-
genes. perature and pressure contain equal numbers of molecules.
aquaporin 28kDa protein, tetramers of which form water chan- One mole (mol) of an ideal gas at 0°C and 1standard atmos-
nels in membranes. phere (atm) occupies 22.414 liters (L). Avogadro's number
area centralis (fovea)In the mammalian retina, the area with the equals 6.02252 X molecules/mol.
highest visual resolution due to the small divergence and con- axon The elongated cylindrical process of a nerve cell along which
vergence in the pathway linking photoreceptors to ganglion action potentials are conducted; a nerve fiber.
cells; in primates contains closely packed cone cells. axonemme Complex of microtubules and associated structures
arginine phosphate A pho~phor~lated nitrogenous compound within the flagellar or ciliary shaft.
axon hillock The transitional region between an axon and the blubber A fatty, insulating tissue typically found under the skin in
nerve cell body. cetaceans.
axon terminal The end of each axon, which typically is the site Bohr effect (Bohr shift) A change in hemoglobin-oxygen affinity
where signals are passed to another neuron or other cell. due to a change in pH.
axoplasm The cytoplasm within an axon. Bolus A discrete plug or collection of food material moving
azide Any compound bearing the N,- group. through the alimentary canal.
bombykol Sex attractant pheromone of the female silkworm moth
(Bombyx mori).
b-adrenergic receptors Epinephrine-binding sites; normally cou- book lungs The respiratory surface in spiders.
pled to activation of adenylate cyclase. Bowman's capsule (glomerular capsule) A globular expansion
baroreceptor A sensory nerve ending that is stimulated by changes at the beginning of a renal tubule and surrounding the
in pressure, as those in the walls of blood vessels. glomerulus.
basal body (kinetosome) Microtubular structure from which a cil- Boyle's law At a given temperature, the product of pressure and
ium or flagellum arises; homologous with centriole. volume of a given mass of gas is constant.
basal cell A cell in a chemoreceptive organ that regularly gives rise bradycardia A reduction in heart rate from the normal level.
to new chemoreceptor cells during adult life. bradykinin A hormone formed from a precursor normally circu-
basal metabolic rate (BMR) The rate of energy conversion in a lating in the blood; a very potent cutaneous vasodilator.
homeotherm while it is resting quietly within the thermal neu- brain The major neuronal center within the body; typically located
tral zone without food in the intestine. at the anterior of the body.
base Proton acceptor. brain hormone (prothoracicotropin; activating hormone) A hor-
basilar membrane The delicate ribbon of tissue bearing the audi- mone synthesized by the neurosecretory cells of the pars in-
tory hair cells in the cochlea of the vertebrate ear. tercerebralis and released by the corpora cardiaca of insects;
Bell-Magendie rule The dorsal root of the spinal cord contains activates the prothoracic glands to secrete ecdysone.
only sensory axons, whereas the ventral root contains only bronchi Conducting airways in the lung; branches of the tracheae.
nlotor axons. bronchioles Small conducting airways in the lung; branches of the
Bernoulli's effect Fluid pressure drops as fluid velocity increases. bronchi.
beta (b)adrenergicreceptors The class of adrenergic membrane re- brood spot A prolactin-induced bald area on the ventral surface of
ceptors that are blocked by the drug propranolol; their acti- some brooding birds that receives a rich supply of blood for
vation is less sensitive to norepinephrine than is that of the the incubation of eggs.
alpha receptors. brown fat Fat deposits with extensive vascularization, mitochon-
beta keratin Insoluble, sulfur-rich scleroprotein;constituent of epi- dria and enzyme systems for oxidation. Found in small, spe-
dermis, horns, hair, feathers, nails, and tooth enamel. Beta cific deposits in a few mammals and used for nonshivering
refers to the protein's secondary structure, which is in pleated thermogenesis.
sheets. Brunner's glands Exocrine glands that are located in the intesti-
beta pleated sheet Form of protein secondary structure in which nal mucosa and secrete an alkaline mucoid fluid.
two or more distinct amino acid chains lay side by side, held brush border A free epithelial cell surface bearing numerous mi-
together by hydrogen bonds. crovilli.
bile Viscous yellow or greenish alkaline fluid produced by the liver BTPS Body temperature, atmospheric pressure, saturated with
and stored in the gallbladder; containing bile salts, bile pig- water vapor.
ments, and certain lipids, it is essential for digestion of fats. buccal Pertaining to the mouth cavity.
bile duct Duct carrying bile fluid from liver to duodenum. buffer A chemical system that stabilizes the concentration of a sub-
bide fluid Fluid secreted by liver that emulsifies fats and neutralizes stance; acid-base systems serve as pH buffers, preventing large
acid intestinal contents. changes in hydrogen ion concentration.
bile pigments Pigments in bile fluid derived from breakdown prod- bundle of His The conducting tissue within the interventricular
ucts of hemoglobin. septum of the mammalian heart.
bile salt Bile acid such as cholic acid conjugated with glycine or bungarotoxin (BuTX) A blocking agent composed of a group of
.taurine, promoting emulsification and solubilization of in- neurotoxins isolated from the venom of members of the snake
testinal fats. genus Bungarus (the krait) of the cobra family; binds selec-
binocular convergence Positioning of the eyes so that the images tively and irreversibly to nicotinic acetylcholine receptors.
formed fall on analogous portions of the two retinas, avoid- Bunsen solubility coefficient The quantity of gas at STPD that will
ing double vision. dissolve in a given volume of liquid per unit partial pressure
biogenic amine Any of a number of signalling molecules that are of the gas in the gas phase. This coefficient is used only for
synthesized in the body from single amino acid molecules. gases that do not react chemically with the solvent.
bipolar cell A neuron with two axons emerging from opposite bursicon A hormone secreted by neurosecretory cells of the insect
sides of the soma; one class of these neurons is found in the central nervous system; tans and hardens the cuticle of freshly
vertebrate retina, where they transmit signals from the pho- molted insects.
toreceptor cells to the retinal ganglion cells.
birefringence Double refraction; the ability to pass preferentially cable properties The passive electrical properties (resistance and
light that is polarized in one plane. capacitance) of a cell; these properties were first worked out
bleaching Fading of photopigment color upon absorption of light. for submarine cables.
GLOSSARY G-5
......................................
calcitonin (thyrocalcitonin) A protein hormone secreted by the achieved by membrane embedded carrier (e.g., facilitated
mammalian parafollicular cells of the thyroid in response to diffusion).
elevated plasma calcium levels. carrier molecules Lipid-soluble molecules that act within biologi-
calcitriol A steroid-like compound produced from vitamin D in- cal membranes as carriers for certain molecules that have
gested with some foods and from vitamin D3, or synthesized lower mobility in the membrane.
from cholesterol in the skin. The physiological actions of cal- catabolism Disassembly of complex molecules into simpler ones.
citriol are similar to those of parathyroid hormone. catalysis An increase in the rate of a chemical reaction promoted
calcium response A graded depolarization due to a weakly regen- by a substance-the catalyst-not consumed by the reaction.
erative inward calcium current. catalyst A substance that increases the rate of a reaction without
caldesmon A calcium-binding regulatory protein in smooth mus- being used up in the reaction.
cle, which plays a role in the "latch" mechanism of some cataract A condition in which proteins of the lens of the eye be-
smooth muscles. come cross-linked, causing it to become cloudy and reducing
calmodulin A troponinlike calcium-binding regulatory protein visual acuity.
found in essentially all tissues. catecholamines A group of related compounds exerting a sympa-
calorie (cal)The quantity of heat required to raise the temperature thomimetic action on nervous tissue; examples are epineph-
of 1g of water from 14.5" to 15.S°C; most commonly used as rine, norepinephrine, and dopamine.
kilocalorie (kcal) = 1000 cal. catecholandues A group of related compounds exerting a sympa-
calorimetry Measurement of heat production in an animal. thomimetic action on nervous tissue; examples are epineph-
calsequestrin A calcium-binding protein that contributes to the rine, norepinephrine, and dopamine.
regulation of contraction in smooth muscles. cathode The negative electrode, so called because it is the electrode
canines Pointed, dagger-like teeth are used for piercing and tear- to which cations are attracted.
ing food. cation A positively charged ion; attracted to a negatively charged
capacitance The property of storing electric charge by electrosta- electrode.
tic means. caudal Pertaining to the tail end.
capacitive current Current entering and leaving a capacitor. cecum A blind pouch in the alimentary canal.
capacity The ability of a capacitor or other body to store electric
Cellulase An enzyme that digests cellulose and hemicellulose, pro-
charge. The unit of measure is the farad (F), which describes
duced by gut-borne symbiotic micro-organisms.
the proportionality between charge stored and potential for a
central chemoreceptors Sensory structures in the brain monitoring
given voltage, C = q N = coulombs divided by volts.
pH and initiating appropriate changes in breathing.
carbohydrases Enzymes that specifically break down carbohy-
central lacteal Small, blind-ended lymph vessel in center of in-
drates.
testinal villus for the uptake of fats and some vitamins.
carbohydrate Aldehyde or ketone derivative of alcohol; utilized by
animal cells primarily for the storage and supply of chemical central nervous system The collection of neurons and parts of neu-
energy; most important are the sugars and starches. rons that are contained within the brain and spinal cord of
vertebrates, or within the brain, ventral nerve cord, and ma-
carbonic anhydrase An enzyme catalyzing the reversible intercon-
jor ganglia of invertebrates.
version of carbonic acid to carbon dioxide and water.
central pattern generator A group of neurons that produces and
carbonyl The organic radical-C=O,which occurs in such com-
maintains a rhythmic pattern of action, such as breathing,
pounds as aldehydes, ketones, carboxylic acids, and esters.
walking, chewing, or swimming.
carboxyhemoglobin The compound formed when carbon monox-
ide combines with hemoglobin; carbon monoxide competes central pattern-generating network A set of interconnected neu-
successfully with oxygen for combination with hemoglobin, rons whose collective activity produces patterned behavior.
producing tissue anoxia. central sulcus A deep, almost vertical furrow on the cerebrum,
carboxylates R-COO-, salts or esters of carboxylic acids. dividing the frontal and parietal lobes.
carboxyl group The radical -COOH, which occurs in the car- cephalic Pertaining to the head.
boxylic acids. cephalic phase Gastric secretion occurring in response to sight,
cardiac output The total volume of blood pumped by the heart per smell, and/or taste of food, or in response to conditioned
" unit of time; cardiac output equals heart rate times stroke vol- reflexes.
ume. cephalization The evolutionary tendency for the neurons of higher
carotid body A nodule on the occipital artery just above the organisms to be concentrated in a brain located at the anterior
carotid sinus, containing chemoreceptors that sense the chem- end of the animal.
ical composition of arterial blood. cerebellum A part of the hindbrain that contributes to the coor-
carotid rete Countercurrent heat exchanger at base of skull that dination of motor output.
helps prevent overheating of the brain of certain hoofed ani- cerebral cortex The thin layer of gray matter that covers the cere-
mals and carnivores. brum of mammals, and probably of birds.
carotid sinus A dllat~onof the Internal carot~dartery wlth many cerebral hemispheres The large paired structures of the cerebrum,
baroreceptor (pressure receptor) endlngs In the wall. connected by the corpus callosum.
carotid sinus baroreceptors Receptors that sense arter~alblood cerebral ventricles A series of confluent fluid-filled cavities within
pressure; located in the carot~dsmus, a dllatat~onof the Inter- the brain of vertebrates; the fluid in the ventricles is cere-
nal carot~dartery at ~ t orrgln.
s brospinal fluid.
carrier-mediated transport Transmembrane transport of solutes cerebrospinal fluid In vertebrates, clear fluid that fills the cavities
G-6 GLOSSARY
......................................
(ventricles)within the brain and the central canal of the spinal cells from the digestion produces of fats, monoglycerides,
cord; it is a complex filtrate of blood plasma and is modified fatty acids, and glycerol.
by brain cells before it returns to the venous system. chyme The mixture of partially digested food and digestive juices
cerebrum The largest part and highest center of the mammalian found in the stomach and the intestine.
brain; it evolved from the olfactory centers of lower verte- chymotrypsin A proteolytic enzyme that specifically attacks pep-
brates. tide bonds containing the carboxyl groups of tyrosine, phenyl-
charge, electric ( q ) Measured in units of coulombs (C). To alanine, tryptophan, leucine, and methionine.
convert 1 g .equiv weight of a monovalent ion to its ele- chymotrypsinogen Inactive precursor of chymotrypsin.
mental form (or vice versa) requires a charge of 96,500 C
ciliary body A thick region of the anterior vascular tunic of the eye;
(1faraday, F). Thus, in loose terms, a coulomb is equivalent
joins the choroid and the iris.
to 1196,500 g .equiv of electrons. The charge on one electron
ciliary muscle A muscle of the ciliary body of the vertebrate eye;
is - 1.6 x C. If this is multiplied by Avogadro's number,
the total charge is 1 F, or -96,487 C . molk'. influences the shape of the lens in visual accommodation.
chelating agent A chemical that binds calcium, or other ions, and ciliary reversal A change in the direction of the power stroke of a
removes it from solution. cilium, causing it to beat in reverse.
chemical energy Energy contained in the chemical bonds holding cilium A motile organelle with a "9 + 2" tubular substructure;
molecules together. when present generally in large numbers, a small flagellum.
chemical synapse A junction between a neuron and another cell in circadian rhythms Biological rhythms with daily cycles.
which the signal from the presynaptic neuron is carried across circalunar Referring to biorhythms related to lunar cycles.
the synaptic cleft by neurotransmitter molecules. circannual Referring to biorhythms related to yearly cycles.
chemoreceptor A sensory receptor specifically sensitive to certain circatidal Referring to biorhythms related to tidal cycles.
molecules. circular smooth muscle Inner layer of smooth muscle that encir-
chief cells (zygomatic)Ep~thelialcells of the gastric epithelium that cles small intestine.
release pepsin. cis A configuration with similar atoms or groups on the same side
chimera An animal with extra copies of a normal gene or a copy of the molecular backbone.
of a mutated gene.
cis-trans isomerization Conversion of a cis isomer into a trans
chitin A structural polymer of D-glucosamme that serves as the isomer.
primary constituent of arthropod exoskeletons.
citric acid cycle (also Krebs cycle, TCA cycle) A series of eight
chloride cells Epithelial cells of fish gills that engage in active trans-
major reactions following glycolysis, in which acetate residues
port of salts.
are degraded to CO, and H20.
chloride shift The movement of chloride ions across the red blood
cladogram A form of diagrammatic analysis of taxonomic rela-
cell membrane to compensate for the movement of bicarbon-
tionships between organisms that relates animals according to
ate ions.
suites of common characters.
chlorocruorin A green respiratory pigment found in some marine
clathrin The protein surrounding the cytoplasmic surface of a
polychaetes; similar to hemoglobin.
coated vesicle membrane.
choanocytes Flagellated cells lining the body cavity of sponges.
clefts Lateral intercellular spaces between cells in epithelium.
cholecystokinin (CCK, pancreozymin; CCK-PZ) A hormone lib-
erated by the upper intestinal mucosa that induces gallblad- cloaca The terminal area of hindgut of some fishes, amphibians,
der contraction and release of pancreatic enzymes. reptiles, birds and a few mammals; aids in urinary ion and wa-
cholesterol A natural sterol; precursor to the steroid hormones. ter resorption.
cholinergic Relating to acetylcholine or substances with actions clone A population of genetically identical cells all derived from
similar to ACh. a single original cell.
choroidplexus Highly vascularized, furrowed projections into the coated pits Receptor-lined membrane depressions that eventually
brain ventricles that secrete cerebrospinal fluid. form coated vesicles during the process of receptor-mediated
choroid rete A countercurrent arrangement of arterioles and endocytosis.
venules behind the retina in the eyes of teleost fish. coated vesicle Vesicle with its cytoplasmic surface covered with the
chromaffin cells Epinephrine-secreting cells of the adrenal protein clathrin, formed in process of receptor-mediated
medulla; named for their high affinity for chromium salt endocytosis.
stains. cochlea A portion of the inner ear, a tapered tube would into a
chromatography A general technique that exploits the fact that spiral like the shell of a snail, containing hair cell receptors for
different components in a sample will move at different rates detecting sound.
through a substrate such as chromatography paper or other cochlear microphonics Electrical signals recorded from the
porous solid matrices. cochlea, having a frequency identical to that of the sound
chromophore A chemical group that lends a dist~nctcolor to a stimulus.
compound containing it. coelenteron A blind-ended tube or cavity in coelenterates that
chronobiology The study of biological rhythms. serves as a "batch reactor" site for chemical digestion.
chronotropic Pertaining to rate or frequency, especially in refer- coelom The body cavity of higher metazoans, situated between the
ence to the heartbeat. gut and the body wall and lined with mesodermal epithelium.
chylomicrons Protein-coated tiny droplets of triglycerides, phos- coenzyme An organic molecule that combines with an apoenzyme
pholipids and cholesterol formed within vesicles of absorptive to form the functioning holoenzyme.
GLOSSARY G-7
......................................
coenzyme A (CoA) A derivative of pantothenic acid to which ac- contralateral Pertaining to the opposite side.
etate becomes attached to form acetyl CoA. conus A chamber invested with cardiac muscle and found in series
cofactor An atom, ion, or molecule that combines with an enzyme with and downstream from the ventricle in elasmobranchs.
to activate it. convection The mass transfer of heat due to mass movement of a
coitus Sexual intercourse. gas or liquid.
colchicine An antimitotic agent that disrupts microtubules by in- convergence A pattern in which inputs from many different neu-
terfering with the polymerization of tubulin monomers. rons impinge upon a single neuron.
collar cells Flagellated cells h i n g the internal chambers of sponges corepressor A low-molecular-weightmolecule that unites with an
(Porifera). aporepressor to form a substance that inhibits the synthesis of
collateral processes Branches of an axon that terminate in loca- an enzyme.
tions other than the major target location. cornea The clear surface of the eye through which light passes as
collaterals Side branches of a nerve or blood vessel. it enters the eye.
collecting duct The portion of the renal tubules in which the final corneal lens The clear structure at the outside surface of an om-
concentration of urine occurs. matidium; it admits and focuses light entering the ommatid-
colligative properties Characteristics of a solution that depend on ium.
the number of molecules in a given volume. corpora allata Nonneuronal insect glands existing as paired or-
colloid A system in which fine solid particles are suspended in a gans or groups of cells dorsal and posterior to the corpus car-
liquid. diaca; the corpora allata secrete juvenile hormone (JH).
command system A set of neurons that, when stimulated, elicit a corpora cardiaca Major insect neurohemal organs existing as
set pattern of coordinated movements. paired structures immediately posterior to the brain; they lib-
erate brain hormone.
competitive inhibition Reversible inhibition of enzyme activity due
corpus luteum The yellow ovarian glandular body that arises from
to competition between a substrate and an inhibitor for the
active site of the enzyme. a mature follicle that has released its ovum; it secretes proges-
terone. If the ovum released has been fertilized, the corpus lu-
compliance The change in length or volume per unit change in the
teum grows and secretes during gestation; if not, it atrophies
applied force.
and disappears.
compound eye The multifaceted arthropod eye; the functional unit
cortex External or surface layer of an organ.
is the ommatidium.
corticospinal tract A group of axons of neurons whose somata and
concentration gradient Difference in solute concentration across a
dendrites are located in the motor cortex and whose axon ter-
membrane, or between two different regions of a solution.
minals synapse on motor neurons in the spinal cord.
condensation A reaction between two or more organic molecules
corticotropin A hormone released by cells in the adenohypophysis
leading to the formation of a larger molecule and the elimi-
that acts mainly on the adrenal cortex, stimulating growth
nation of a simple molecule, such as water or alcohol.
and corticosteroid production and secretion in that organ.
conditioned reflex Reflexes learned or modified through behav-
cortisol A steroid hormone secreted by the adrenal cortex.
ioral repetition.
cotransmitter A second neurotransmitter molecule synthesized in
conductance, electrical (G)A measure of the ease with which a
and released from an axon terminal, along with a small trans-
conductor carries an electric current; the unit is the siemen (S), mitter molecule such as acetylcholine or GABA.
re-ciprocal of the ohm (a).
cotransport Carrier-mediated transport in which two dissimilar
conductance, thermal A quantity describing the ease with which molecules bind to two specific sites on the carrier molecule for
heat flows by conduction under a temperature gradient across transport in the same direction.
a substance or an object.
coulomb (C)MKS unit of electric charge; equal to the amount of
conductivity The intrinsic property of a substance to conduct elec- charge transferred in 1 second (s) by 1 ampere (A) of current.
tric current; reciprocal of resistivity. See also charge, electric.
conductor A material that carries electric current. counter current heat exchanger A specialized parallel arrangement
cone A vertebrate visual receptor cell that has a tapered outer seg- of incoming arteries and outgoing veins forming a heat ex-
ment in which the lamellar photosensitive membranes remain changer that conserves heat in the body core.
'
continuous with the surface membrane. countercurrent multiplier A pair of opposed channels containing
confocal scanning microscope A microscope using a focused laser fluids flowing in opposed directions and having an energetic
beam to rapidly scan different areas of the specimen in a sin- gradient directed transversely from one of the channels into
gle plane. Light reflected from that plane is assembled by a the other. Since exchange due to the gradient is cumulative
computer into a composite image of the specimen. with distance, the exchange per unit distance will be multi-
conformer An animal in which internal body fluid condition tends plied, so to speak, as a function of the total distance over
to parallel that of the external environment. which exchange takes place.
conjugate acid-base pair Two substances related by gain or loss of counterion An ion associated with, and of opposite charge to, an
an H+ ion (proton). ion or an ionized group of a molecule.
connective A collection of axons that carry information between countertransport The uphill membrane transport of one substance
neuronal centers, such as ganglia, in many invertebrate ner- driven by the downhill diffusion of another substance.
vous systems. coupled transport Uptake of one substance into a cell that depends
contracture A more or less sustained contraction in response to an on the downhill diffusion exit of another substance into the
abnormal stimulus. cell.
G-8 GLOSSARY
.........................................
covalent bond A bond formed by electron-pair sharing between cytoplasm The semifluid substance within a cell, exclusive of the
two atoms. nucleus but including other organelles.
creatine phosphate (phosphocreatine)A phosphorylated nitroge- cytosine Oxyamino-pyrimidine, C,H,N,O; base component of
nous compound found primarily in muscle; acts as a storage nucleic acid.
form of high-energy phosphate for the rapid phosphorylation cytosol The unstructured aqueous phase of the cytoplasm between
of ADP to ATP. the structured organelles.
creatinine Nitrogenous waste product of muscle creatine.
cretinism A chronic condition caused by hypothyroidism in child- D600 Methoxyverapamil; an organic drug that blocks calcium
hood; characterized by arrested physical and mental develop- influx through cell membranes.
ment. D-tubocurarine See curare.
cristae The folds of the inner mitochondria1 membrane. Dalton's law The partial pressure of a gas in a mixture is indepen-
critical fusion frequency The number of light flashes per second at dent of other gases present. The total pressure is the sum of
which the light is perceived to be continuous. the partial pressures of all gases present.
critical thermal maximum The temperature above which long- dark current Steady sodium current that leaks into a vertebrate vi-
term survival is not possible. sual receptor cell at the outer segment. The sodium is actively
pumped out of the inner segment, completing the circuit. The
crop A muscular organ in the upper alimentary canal of birds; re-
dark current is reduced by photoexcitation.
ceives swallowed food items and small stones, and churns the
decerebration Experimental elimination of cerebral activity by sec-
mixture together to break down the food into more digestible
tion of the brain stem or by interruption of the blood supply
particles.
to the brain.
crop milk A nutrient-rich substance fed by regurgitation to pigeon
decussation Crossing over from one side to the other.
chicks by both parents.
defecation The process of expelling feces.
cross-bridges Spirally arranged projections from myosin thick fil-
dehydrogenase An enzyme that "loosens" the hydrogen of a sub-
aments that bind to sites on actin thin filaments during mus-
strate in preparation for passage to a hydrogen receptor.
cle contraction.
dehydroretinal An aldehyde of dehydroretinol.
crustecdysone A steroid hormone that promotes molting in crabs.
dehydroretinol (retinol2; vitamin A2) The form of vitamin A oc-
crypt of Lieberkiihn A circular depression around the base of each
curring in the liver and the retina of freshwater fishes, some
villus in the intestine.
invertebrates, and amphibians.
cupula A small upside-down cup or domelike cap; in the lateral- delayed outward current (late outward current) Current carried
line and the vertebrate organs of equilibrium, the cupula cov- by Kt through channels that open with a time lag after onset
ers hair cells in a gelatinous matrix. of a depolarization; responsible for repolarization of the ac-
curare (D-tubocurarine) South American arrow poison; blocks tion potential.
synaptic transmission at the motor endplate by competitive in- denaturation Alteration or destruction of the normal nature of a
hibition of nicotinic acetylcholine receptors. substance by chemical or physical means.
current, electric The flow of electric charge. A current of 1 dendrites Fine processes of a neuron, often providing the main
coulomb (C) per second is called an ampere (A). By conven- receptive area of the cell onto which synaptic contacts are
tion the direction of current flow is the direction that a posi- made.
tive charge moves (i.e., from the anode to the cathode). densitometer An instrument that measures the amount of expo-
cuticle The hard outer coat of insects and crustacea secreted by an sure of the film emulsion produced by autoradiography.
epidermal layer, the hypodermis. deoxyhemoglobin Hemoglobin in which oxygen is not combined
cyanide A compound containing cyanogen and one other ele- to the Fe,, of the heme moiety.
ment; blocks transfer of electrons from the terminal cy- deoxyribonucleic acid See DNA.
tochrome a and a, to oxygen in the respiratory chain. depolarization The reduction or reversal of the potential difference
cybernetics The science of information communication and con- that exists across the cell membrane at rest.
trol in animals and machines. desmosome A type of cell junction serving primarily to aid the
cyclic AMP (CAMP)A ubiquitous cyclic nucleotide (adenosine structural bonding of neighboring cells.
3'5'-cyclic monophosphate) produced from ATP by the en- diabetes mellitus A metabolic malady in which there is a partial or
zymatic action of adenylate cyclase; important cellular regu- complete loss of activity in the pancreatic islets; the concomi-
latory agent that acts as the second messenger for many tant insulin insufficiency leads to inadequate uptake of glucose
hormones and transmitters. into cells and loss of blood glucose in the urine.
cyclic GMP (cGMP) A cyclic nucleotide (guanosine 3'5'-cyclic diacylglycerol (DAG) A diglyceride, present as a constituent of
monophosphate) analogous to CAMP but present in cells at cellular phospholipids; when released from these phospholip-
a far lower concentration and producing target cell responses ids by an agonist-activated phospholipase, this molecule
that are usually opposite to those of CAMP. serves as the endogenous activator of the calcium- and
cyclostomes A group of jawless vertebrates, including lampreys calmodulin-dependent protein kinase (protein kinase C) and
and hagfishes. is part of an important intracellular signalling cascade.
cytochalasin A drug that disrupts cytoplasmic microfilaments. dialysis The process by which crystalloidsand macromolecules are
cytochromes A group of iron-containing proteins that function in separated by utilizing the differences in their diffusion rates
the electron-transport chain in aerobic cells; they accept and through a semipermeable membrane.
pass on electrons. diaphragm The dome-shaped muscle that separates the thoracic
and abdominal cavities and serves as the chief muscle of res- when two solutions are separated by a membrane permeable
piration. to only some of the ions of the solutions.
diastole The phase in the heartbeat during which the myocardium dopanitine (hydroxytyramine) A product of the decarboxylation
is relaxed and the chambers are filling with blood. of dopa, an intermediate in norepinephrine synthesis; a cen-
dielectric constant A measure of the degree to which a substance tral nervous system transmitter.
is able to store electric charge under an applied voltage; the di- dormancy The general term for reduced body activities, including
electric constant of a material depends on charge distribution sleep, torpor, hibernation, "winter sleep," and estivation.
within molecules. dorsal horn The dorsal part of the gray matter in the vertebrate
diffusion Dispersion of atoms, molecules, or ions as a result of ran- spinal cord; contains the cell bodies of neurons that receive,
dom thermal motion. process, and transmit sensory information.
diffusion coefficient Coefficient relating rate of diffusional flux to dorsal root A nerve trunk that enters the spinal cord near the dor-
concentration gradient, path length, and area across which sal surface; contains sensory axons only.
diffusion occurs. dorsal root ganglion (DRG) On the surface of the dorsal root, a
digastric stomach The multichambered stomach of ruminants. collection of the cell bodies of sensory neurons that send
digestive enzymes Enzymes secreted by alimentary canal to aid in processes into the region of the body that is innervated by that
chemical digestion. spinal segment; each spinal segment contains bilaterally
paired DRGs.
digestive epithelium Epithelium lining the small intestine.
drag The resistance to movement of an object through a medium,
dimer A molecule made by the joining of two molecules (i.e.,
increasing with the viscosity and density of the medium and
monomers) of the same kind.
the surface area and shape of the object.
dinitrophenol (DNP) Any of a group of six isomers,
ductus arteriosus The vessel connecting the pulmonary artery and
C,H,(OH)(NO,), ,that act as aerobic metabolic inhibitors by
the aorta in fetal mammals.
virtue of their ability to uncouple oxidation from phospho-
duodenum The initial section of the small intestine, situated be-
rylation in mitochondria1 electron transport.
tween the pylorus of the stomach and the jejunum.
dipole A molecule with separate regions of net negative and net
dwarfism An abnormally small size in humans; a result of insuf-
positive charge, so that one end acts as a positive pole and the
ficient growth hormone secretion during childhood and
other as a negative pole.
adolescence.
dipole moment The electrostatic force required to align a dipolar
dynamic range The range of energy over which a sensory system
molecule parallel to the electrostatic field; the force required
is responsive and can encode information about stimulus in-
increases as the separation of the molecular charges decreases.
tensity.
disaccharide sugars A double sugar formed when two monosac- dynein A ciliary protein with magnesium-activated ATPase
charides (single sugars) are joined together by dehydration
activity.
synthesis.
dynein arms Projections from tubule A of one microtubule dou-
disinhibition Release of a neuron from inhibitory input when the blet toward tubule B of the next, composed of a protein ex-
inhibitory neuron is, itself, synaptically inhibited. hibiting ATPase activity.
dissociation Separation; resolution by thermal agitation or solva- dyspnea Labored, difficult breathing.
tion of a substance into simpler constituents.
dissociation constant K' = [H][A-]/[HA]. The empirical measure
of the degree of dissociation of a conjugate acid-base pair in
early inward current Depolarizing current of excitable tissues, car-
solution.
ried by Na+ or Ca2+;responsible for the upstroke of the ac-
distal More distant from a point of reference in the centrifugal di- tion potential.
rection; i.e., away from the center. early receptor potential (ERP)An almost instantaneous potential
distal tubule The renal tubules located in the renal cortex leading change recorded from the retina in response to a short flash of
from (and continuous with) the ascending limb of the loop of light that probably corresponds to a movement of charge that
Henle to the collecting duct. occurs as the photopigment undergoes conformational
distance of closest approach Shortest possible span between the change.
centers of two atoms. eccentric cell In Limulus, the afferent neuron of each ommatidium;
disulfide linkage Bond between sulfide groups that determines pro- it is surrounded by and receives information from photore-
tein tertiary structure by linking together portions of polypep- ceptive retinular cells.
tide chains. ecdysis Shedding of the outer shell; molting in an arthropod.
diuresis An increase in urine excretion. ecdysone A steroid hormone secreted by the thoracic gland of
diuretic An agent that increases urine secretion. arthropods that induces molting.
divalent Carrying an electric charge of two units; a valence of 2. echolocation The ability of some species to recognize and locate
divergence A pattern in which the axon of a single neuron objects in their environment by emitting sounds and receiving
branches, allowing it to synapse onto more than one synaptic the sound that is reflected back to them by the objects.
target. eclosion hormone Insect hormone that induces the emergence of
DNA (deoxyribonucleic acid) The class of nucleic acids responsi- the adult from the puparium.
ble for hereditary transmission and for the coding of amino ectopic pacemaker A pacemaker situated outside the area where it
acid sequences of proteins. is normally found.
Donnan equilibrium Electrochemical equilibrium that develops ectoplasm The gel-state cytoplasm surrounding the endoplasm.
G-10 GLOSSARY
.........................................
ectotherms Refers to animals whose body temperature is depen- from embryonic muscle and uses synapses similar to neuro-
dent on heat in the environment. muscular junctions to build up the charge.
edema Retention of interstitial fluid in organs or tissues. electroreceptors Sensory receptors that detect electrical fields.
EDTA Ethylenediaminetetraacetic acid; a calcium- and electroretinogram (ERG) An extracellular electrical recording,
magnesium-chelating agent. made at the surface of the eye, of activity in many visual re-
effector A cell, tissue, or organ that acts to change the condition of ceptors and other retinal neurons.
an organism (e.g., by contracting muscles or secreting a hor- electrotonic potential A graded potential generated locally by cur-
mone) in response to neuronal or hormonal signals. rents flowing across the membrane; not actively propagated
efferent (nerve or neuron) Centrifugal; a neuron that carries in- and not all-or-none.
formation from higher brain centers toward structures in the Embden-Meyerhof pathway See glycolysis.
periphery. endergonic Characterized by a concomitant absorption of
efflux Movement of solute or solvent out of a cell across the cell energy.
membrane. endocardium The internal lining of the heart chamber.
EGTA Ethyleneglycol-bis(p-aminoethylether)-N,N'-tetraacetic endocrine A hormonal pathway characterized by the production
acid; a calcium chelating agent. of a biologically active substance by a ductless gland; the sub-
elastic Capable of being distorted, stretched, or compressed with stance then is carried through the bloodstream to initiate a cel-
subsequent spontaneous return to original shape; resilient. lular response in a distal target cell or tissue.
electrical potential Electrostatic force, analogous to water pres- endocrine glands Ductless organs or tissues that secrete a hormone
sure; a potential difference (i.e., voltage) is required for the into the circulation.
flow of electrical current across a resistance. endocytosis Bulk uptake of material into a cell by membrane in-
electrical synapse A junction between two cells at which a signal pocketing to form an internal vesicle.
is carried from one cell to the other by the passage of charged endogenous Arising within the body.
particles through gap junctions. endogenous opioids Neurotransmitter or neuromodulator mole-
electrocardiogram (ECG) The record of electrical events associ- cules (e.g., endorphins and enkephalins) whose receptors bind
ated with contractions of the heart; obtained with electrodes the opioid drugs, such as opium and heroin; in some parts of
placed on other portions of the body. the nervous system, when these transmitters bind to their re-
electrochemical equilibrium The state at which the concentration ceptor molecules, the net result is reduction in the perception
gradient of an ion across a membrane is precisely balanced by of pain.
the electric potential across the membrane. endolymph Aqueous liquid with a high K+ concentration and a
electrochemical gradient The sum of the combined forces of con- low Na+ concentration found in the vertebrate organs of equi-
centration gradient and electrical gradient acting on an ion. librium and in the scala medza of the cochlea.
electrochemical potential The electrical potential developed across endometrium An epithelium that lines the uterus.
a membrane due to a chemical concentration gradient of an endopeptidases Proteolytic enzymes that break up large peptide
ion that can diffuse across the membrane. chains into shorter polypeptide segments.
electrode An electrical circuit element used to make contact with endoplasm The sol-state cytoplasm that streams within the cell in-
a solution, a tissue, or a cell interior; used either to measure terior.
potential or to carry current. endorphins Endogenous neuropeptides that exhibit morphinelike
electrogenic Giving rise to an electric current or voltage. actions, found in the central nervous system of vertebrates;
electrolyte A compound that dissociates into ions when dissolved different types consist of 16,17, or 31 amino residues.
in water. endothelium Single cell layer forming the internal lining of blood
electromotive force (emf) The potential difference across the ter- vessels.
minals of a battery or any other source of electric energy. endotherms animals whose body temperature is dependent on
electronegativity Affinity for electrons. heat production by the body.
electroneutrality rule For a net potential of zero, the positive and endplate A traditional name of the vertebrate neuromuscular
negative charges must add up to zero; a solution must contain synapse, where the motor axon forms many fine terminal
essentially as many anionic as cationic charges. branches that end over a specialized system of folds in the
electron shells Energy levels of electrons surrounding the nucleus. postsynaptic membrane of the muscle cell.
electron-transport chain (respiratory chain) A series of enzymes endplate potential (epp)A postsynaptic potential in the muscle at
that transfer electrons from substrate molecules to molecular the neuromuscular junction (or motor endplate).
oxygen. end-product inhibition (feedback inhibition) Inhibition of a
electro-olfactogram An extracellular electrical recording of the biosynthetic pathway by the end product of the pathway.
summed activity in many olfactory receptors. energy Capacity to perform work.
electro-osmosisMovement of water through a membrane of fixed energy metabolism The complex collection of biochemical reac-
charge in response to a potential gradient. tions within cells that generate ATP and other high-energy
electrophoresis A technique for separating proteins using the net compounds, which serve as the immediate source of energy
positive or negative charge on their surface amino acids. for all biological events.
electroplax organ An organ in electric fishes such as Torpedo that enkephalins Endogenous nuropeptides exhibiting morphinelike
builds up and delivers a powerful electrical charge sufficient actions, found in the central nervous system of vertebrates;
to stun or even to kill prey. The electroplax organ is derived these peptides consist of five amino acid residues.
GLOSSARY G-11
........................................
enterogastric reflex A reflex that inhibits gastric secretion, trig- excitability The property of altered membrane conductance (and
gered when duodenum is stretched by chyme pumped from often membrane potential) in response to stimulation.
the stomach. excitation-contraction coupling In muscle fibers, the process by
enterogastrone A hormone that is secreted from the duodenal mu- which electrical excitation of the surface membrane leads to
cosa in response to fat ingestion and that suppresses a gastric activation of the contractile process.
motility and secretion. excitation-secretion coupling Sequence of molecular events re-
enterokinase Intestinal proteolytic enzyme. sponsible for secretion of stored chemicals from an activated
enthalpy The heat produced or taken up by a chemical reaction. secretory cell.
entropy Measure of that portion of energy not available for excitatory In neurophysiology, pertaining to the enhanced prob-
work in a closed system; measure of molecular randomness. ability of producing an action potential.
Entropy increases with time in all irreversible processes. excitatory postsynaptic potential (epsp) A change in the mem-
enzyme A protein with catalytic properties. brane potential of a postsynaptic cell that increases the prob-
enzyme activity A measure of the catalytic potency of an enzyme: ability of an action potential in the cell.
the number of substrate molecules that react per minute per exergonic Characterized by a concomitant release of heat energy.
enzyme molecule. exocrine Of or relating to organs or structures that secrete sub-
enzyme induction Enzyme production stimulated by the specific stances via a duct.
substrate (inducer) of that enzyme or by a molecule struc- exocrine gland A gland that secretes a fluid via a duct.
turally similar to the substrate. exocytosis Fusion of the vesicle membrane to the surface mem-
epicardium The external covering of the heart wall. brane and subsequent expulsion of the vesicle contents to the
epididymis A long, stringlike duct along the dorsal edge of the cell exterior.
testis; its function is to store sperm. exopeptidases Enzymes that attack only peptide bonds near the
epinephrine Generic name for the catecholamine released from the end of a pept~dechain, providing free amino acids, plus dipep-
adrenal cortex; also known by the trade name Adrenalin. tides and tripeptides.
equilibrium The state in which a system is in balance as a result exothermic See exergonic.
of equal action by opposing forces arising from within the expiratory neurons Neurons in the medulla of the brain control-
system. ling actlvity in motor neurons innervating muscles involved in
equilibrium potential The voltage difference across a membrane expiration (breathing out).
at which an ionic species that can diffuse across the membrane explants Small pieces of tissue removed from a donor animal kept
is in electrochemical equilibrium; it is dependent on the con- alive and grown in a flask filled with the appropriate mixture
centration gradient of the ions and is described by the Nernst nutrients.
equation. extensor A muscle that extends or straightens a limb or other ex-
equimolar Having the same molarity. tremity.
equivalent pore size Cell membrane pore diameter that accounts exteroceptors Sense organs that detect stimuli arriving at the sur-
for the rate of diffusion of polar substances across the face of the body from a distance.
membrane. extracellular digestion Digestion occurring outside of the cell in an
eserine (physostigmine)An alkaloid (C,,H,,N,O,) of plant origin alimentary system.
that blocks the enzyme cholinesterase. extrafusal fiber Contractile muscle fibers that make up the bulk of
esophagus The region of the alimentary canal that conducts food skeletal muscle. See also intrafusal fibers.
from the headgut to the digestive areas. extravasation Forcing fluid out of blood vessels, usually blood,
essential amino acids Amino acids that cannot be synthesized by serum, or lymph.
an animal, but are required for synthesis of essential proteins. eye An organ of visual reception that includes optical processing
esterases Enzymes that hydrolyze esters. of light as well as photoreceptive neurons.
estivation Dormancy in response to high ambient temperatures
and/or danger of dehydration. facilitated transport (carrier-mediated transport) Downhill trans-
estradiol-17P The most active natural estrogen. membrane diffusion aided by a carrier molecule that enhances
estrogens A family of female sex steroids responsible for produc- the mobility of the diffusing substance in the membrane.
ing estrus and the female secondary sex characteristics; also facilitation An increase in the efficacy of a synapse as the result of
prepares the reproductive system for fertilization and implan- a preceding activation of that synapse.
tation of the ovum; synthesized primarily in the ovary, al- factorial scope for locomotion A ratio between the basal metabolic
though some is made in the adrenal cortex and male testis. rate and the maximum metabolic rate that can be achieved
estrous cycle Periodic episodes of "heat," or estrus, marked by sex- with intense exercise.
ual receptivity in mature females of most mammalian species. Fahraeus Lindqvist effect The reduction in apparent viscosity of
ethers A class of compounds in which two organic groups are blood as it flows into small arterioles.
joined by an oxygen atom, R1-0-R,. farad (F)The unit of electrical capacitance.
eupnea Normal breathing. faraday Measure of electric charge, -96,487 C .mol-'.
euryhaline Able to tolerate wide variations in salinity. Faraday's constant (F)The equivalent charge of a mole of elec-
exchange diffusion A process by which the movement of one mol- trons, equal to 9.649 X lo4 coulombs (C) per mole of
ecule across a membrane enhances the movement of another electrons.
molecule in the opposite direction; most likely involves a com- fast chemical synaptic transmission Synaptic transmission at a
mon carrier molecule. chemical synapse that is mediated by neurotransmitters that
G-12 GLOSSARY
......................................
bind on the postsynaptic membrane to receptor protein com- energetic (has a longer wavelength) than the light producing
plexes, each of which includes an ion channel. Binding of the the excitation.
transmitter to the receptor complex is sufficient to open (or to flux The rate of flow of matter or energy across a unit area.
close) the ion channel. folds of Kerckring See plica circularis.
feces Undigested material and bacteria eliminated from the follicle-stimulating hormone (FSH) An anterior pituitary go-
hindgut. nadotropin that stimulates the development of ovarian folli-
feedback The return of output to the input part of a system. In cles in the female and testicular spermatogenesis in the male.
negative feedback, the sign of the output is inverted before be- follicular phase That part of the estrous and menstrual cycles that
ing fed back to the input so as to stabilize the output. In pos- is characterized by formation of and secretion by the Graafian
itive feedback, the output is unstable because it is returned to follicles.
the input without a sign inversion, and thus becomes self- food vacuole Enzyme-filled vesicle in Protozoa.
reinforcing, or regenerative. foramen An orifice or opening.
fermentation Enzymatic decomposition;anaerobic transformation foregut The upper region of the alimentary canal involved in food
of nutrients without net oxidation or electron transfer. conduction, storage and digestion.
ferritin A large protein molecule opaque to electrons, used as a Fourier's law The rate of flow of heat in a conducting body is
marker in electron microscopy; normally present in the spleen proportional to its thermal conductance and to the tempera-
as a storage protein for iron. ture gradient.
fever Disease-induced increase in body temperature above normal fovea (area centralis) In the mammalian retina, the area with the
levels. highest visual resolution due to the small divergence and con-
fibrillar muscle Oscillatory insect flight muscle; also termed asyn- vergence in the pathway linking photoreceptors to ganglion
chronous muscle because contractions are not individually cells; in primates, contains closely packed cone cells.
controlled by motor impulses. Frank-Starling mechanism The increase in mechanical work from
fibroblast A connective-tissuecell that can differentiate into chon- the ventricle caused by an increase in end-diastolic volume (or
droblasts, collagenoblasts, and osteoblasts. venous filling pressure).
Fick diffusion equation An equation defining the rate of solute dif- free energy The energy available to do work at a given tempera-
fusion through a solvent. ture and pressure.
field metabolic rate (FMR) The average rate of energy utiliza- frequency modulated The property of a signal in which informa-
tion as the animal goes about its normal activities, which tion is encoded by varying the frequency at which the strength
may range from complete inactivity during resting to maxi- of the signal changes.
mum exertion. fructose A ketohexose, C,H,,O,, found in honey and many fruits.
filter feeding See suspension feeding. fusimotor system The gamma motor neurons and the intrafusal
h a 1 common pathway The concept that the sum total of neuronal fibers that they innervate.
integrative activity expressed in motor output is channeled
through the motor neurons to the muscles.
firing level Potential threshold for the generation of an action gain The increase in signal produced by amplification.
potential. gallbladder The organ associated with liver that concentrates and
first law of thermodynamics Net energy is conserved in any stores bile for eventual discharge into the intestine.
process. gamma aminobutyric acid (GABA) Inhibitory transmitter identi-
first-order enzyme kinetics Describes enzymatic reactions, the rates fied in crustacean motor synapses and in the vertebrate cen-
of which are directly proportional to one reactant's concen- tral nervous system.
tration (either substrate or product). gamma efferents The motor axons innervating the intrafusal mus-
fixed action pattern A behavior that is performed in a stereotyped cle fibers of spindle organs.
fashion in response to specific stimuli. gamma motor neurons Nerve cells of the ventral spinal cord that
flagellum A motile, whiplike organelle similar in organization to a innervate the intrafusal muscle fibers.
cilium, but longer and generally present on a cell only in small gamma rays Electromagnetic radiation of very short wavelength
numbers. (lo-'' cm) and very high energy.
flame cells Flagellated cells at the ending of the excretory collect- ganglia Collections of neuronal cell bodies.
ing tubules of flatworms and nemerteans. ganglion An anatomically distinct collection of neuron cell
flavin adenine dinucleotide (FAD)A coenzyme formed by the con- bodies.
densation of riboflavin phosphate and adenylic acid; it per- ganglion cells A nonspecific term applied to some nerve cell bod-
forms an important function in electron transport and is a ies, especially those located in ganglia of invertebrates or out-
prosthetic group for some enzymes. side the vertebrate central nervous system proper.
flavoproteins Proteins combined with flavin prosthetic groups that ganglion cells (retinal) The afferent neurons that carry visual in-
are important as intermediate carriers of electrons between formation from the vertebrate retina to higher centers of the
the dehydrogenases and cytochromes in the respiratory chain. brain.
flexer A muscle that flexes or bends an extremity. gap junctions Specializationsfor electrical coupling between cells,
fluid mosaic model The accepted model for the cell membrane, in where cell membranes are only about 2 nm apart and tubular
which globular proteins are integrated with the lipid bilayer. assemblies of proteins connect the apposed membranes.
fluorescence The property of emitting light upon molecular exci- gastric Pertaining to the stomach.
tation by an incident light; the emitted light is always less gastric cecum The outpouching of insect alimentary canal,
GLOSSARY G-13
...................................... ........................................
lined with enzyme-secretingand phagocytic cells and serving glutamine Amino acid used, because it is less toxic, to transport
as a stomach. ammonia between the liver and kidneys in mammals.
gastric inhibitory peptide (GIP) A gastrointestinal hormone re- glycocalyx A meshwork of acid mucopolysaccharide and glyco-
leased into the bloodstream from the duodenal mucosa, in- protein filaments that arise from the membrane covering the
hibiting gastric secretion and motility. microvilli of the intestinal brush border.
gastric juice Fluid secreted by the cells of the gastric epithelium. glycogen A highly branched D-glucose polymer found in animals.
gastric phase Secretion phase of digestion stimulated by presence glycogenesis The synthesis of glycogen.
of food in stomach. glycogenolysis The breakdown of glycogen to glucose 6-
gastrin A protein hormone that is liberated by the gastrin cells of phosphate.
the pyloric gland and induces gastric secretion and motility. glycogen synthetase An enzyme that catalyzes the polymerization
gastrointestinal peptide hormones Hormones that regulate the of glucose to glycogen.
basic electric rhythm of smooth muscle in the alimentary glycolipid A lipid containing carbohydrate groups, in most cases
canal. galactose.
Gay-Lussac's law Either the pressure or the volume of a gas is di- glycolysis (Embden-Meyerhof pathway) The metabolic pathway
rectly proportional to absolute temperature if the other is held by which hexose and triose sugars are broken down to sim-
constant. pler substances, especially pyruvate or lactate.
Geiger counter An instrument that detects the presence of ionizing glycosidases Carbohydrases that break down disaccharides (su-
radiation. crose, fructose, maltose, lactose) by hydrolyzing alpha-1,6
gel The stiff, high-viscosity state of cytoplasm. and alpha-1,4 glucosidic bonds into their constituent mono-
gene Regions of coded information carried within the subunits of saccharides for absorption.
DNA forming chromosomes. glycosuria (glucosuria)The excretion of excessive amounts of glu-
generator potential A change in transmembrane potential within cose in the urine.
the receptive part of a sensory neuron; its amplitude is graded goblet cells See mucous cells.
with stimulus intensity, and the potential is conducted eletro- goiter An abnormal increase in size of the thyroid gland, usually
tonically in the neuron. If the potential is sufficiently large at due to a dietary lack of iodine.
the spike-initiating zone of the axon, action potentials will be Goldman equation The equation describing the equilibrium po-
generated. tential for a system in which more than one species of dif-
geniculate body A thalamic nucleus that relays incoming sensory fusible ions are separated by a semi-permeable membrane; if
(auditory and visual) information to the cortex; named for its only one species can diffuse across the membrane the equation
kneelike shape in cross section. reduces to the Nernst equation.
gestation Pertaining to pregnancy. Golgi tendon organs Tension-sensing nerve endings of the Ib af-
gigantism Excessive growth due to hypersecretion of pituitary ferent fibers found in muscle tendons.
growth hormone from birth. gonadotropic hormones (gonadotrophic hormones; go-
gizzard See crop. nadotropins) Hormones that influence the activity of the go-
gland An aggregation of specialized cells that secrete or excrete nads; in particular, those secreted by the anterior pituitary.
substances, such as the pituitary gland, which produces hor- gonadotropins Hormones that act on the gonads.
mones, and the spleen, which takes part in blood production. Graafian follicle A mature ovarian follicle in which fluid is
glial cells (neuroglia)Inexcitable supportive cells associated with accumulating.
neurons in nervous tissue. graded response One that increases as a function of the energy ap-
globin Protein in hemoglobin made of two equal parts each con- plied; a membrane response that is not all-or-none.
sisting of two polypeptide chains. Graham's law The rate of diffusion of a gas is proportional to the
glomerular filtrate rate (GFR) The amount of total glomerular square root of the density of that gas.
filtrate produced per minute by all nephrons of both kidneys; gray matter Tissue of the vertebrate central nervous system con-
equal to the clearance of a freely filtered and nonreabsorbed sisting of cell bodies, unmyelinated fibers, and glial cells.
substance such as insulin. growth hormone (GH, somatotropin) A protein hormone that is
glomerulus A coiled mass of capillaries. secreted by the anterior pituitary and stimulates growth; di-
glucagon A protein hormone released by the alpha cells of the pan- rectly influences protein, fat, and carbohydrate metabolism
creatic islets; its secretion is induced by low blood sugar or by and regulates growth rate.
growth hormone; it stimulates glycogenolysis in the liver. guanine 2-amino-6-oxypurine (CjH,N,jO);a white, crystalline
glucocorticoids Steroids synthesized in the adrenal cortex with base; a breakdown product of nucleic acids.
wide-ranging metabolic activity; included are cortisone, cor- guano White, pasty waste product of birds and reptiles; high in
tisol, corticosterone, and 11-deoxycorticosterone. uric acid content.
gluconeogenesis Synthesis of carbohydrates from noncarbohy- guanosine triphosphate (GTP) A high-energy molecule similar to
drate sources, such as fatty acids or amino acids. ATP that participates in several energy-requiring processes,
glucose 6-carbon sugar comprising the cell's primary metabolic such as peptide bond formation.
fuel; blood sugar. guanylatecydase An enzyme that converts GTP to cyclic GMP.
glutamate A putative excitatory synaptic transmitter in the verte- gustation The sense of taste; chemoreception of ions and
brate central nervous system and in arthropod neuromuscu- molecules in solution by- specialized epithelial sensory
lar junctions. receptors.
G-14 GLOSSARY
......................................
H zone The light zone in the center of the resting muscle sarco- tor]/[H+ donor]). The formula for calculation of the pH of a
mere, where myosin filaments do not overlap with actin fila- buffer solution.
ments; the region between actin filaments. Henry's law The quantity of gas that dissolves in a liquld is nearly
habituation The progressive loss of behavioral response probabil- proportional to the partial pressure of that gas in the gas
ity with repetition of a stimulus. phase.
hair cell A mechanosensory epithelial cell bearing stereocilia and hepatocyte A liver cell.
in some cases a kinocilium. herbivores Animals that feed solely on plant material.
Haldane effect Reduction in total C02content of the blood at con- Hering-Breuer reflex A reflex in which lung inflation activates pul-
stant PCOz when hemoglobin is oxygenated. monary stretch receptors that inhibit further inspiration dur-
half-width The length of time during which a transient physiolog- ing that cycle; activity from stretch receptors is carried in the
ical variable is half of its maximum value or greater. vagus nerve.
halide A binary compound of a halogen and another element. hertz (Hz)Cycles per second.
halogens A family of related elements that form similar saltlike Hess's law The total energy released in the breakdown of a fuel to
compounds with most metals; they are fluorine, chlorine, a given set of end products is always the same, irrespective of
bromine, and iodine. the intermediate chemical steps or pathways used.
headgut The anterior (cranial)region of the alimentary canal pro- heterodimers Dimers consisting of two nonidentical subunits.
viding an external opening for food entry. heterosynaptic facilitation Increased effectiveness of synaptic
heat Energy in the form of molecular or atomic vibration that is transmission between two neurons that occurs as a result of
transferred by conduction, convection, and radiation down activity in a third neuron.
a thermal gradient. heterosynaptic modulation A change in synaptic efficacy at one
synapse that occurs due to activity at another, separate
heat capacity Amount of heat required to raise 1g of substance
synapse. In heterosynaptic facilitation synaptic efficacy in-
1°C.
creases. In heterosynaptic depression, the synaptic efficacy de-
heater tissues Tissues specialized for heat production, e.g. modi-
creases.
fied eye muscles in billfishes.
heterotherm An animal that derives essentially all of its body heat
heat of shortening Thermal energy associated with muscle con-
from the environment.
traction; it is proportional to the distance the muscle has
heterotrophic Depending on energy-yieldingcarbon compounds
shortened.
derived from the ingestion of other plants or animals.
heat of vaporization Heat necessary per mass unit of a given liq-
hexose A six-carbon monosaccharide.
uid to convert all of the liquid to gas at its boiling point.
hibernation A period of deep torpor, or winter dormancy, in ani-
heavy meromyosin (HMM; H-meromyosin) The "head" and
mals in cold climates, lasting weeks or months.
"neck" of the myosin molecule, the part of the myosin mole-
hindgut The terminal region of the alimentary canal, responsible
cule that has ATPase activity.
for storing and eventually eliminating the remnants of di-
helicotrema The opening that connects the scala tympani and the gested food.
scala uestibuli at the cochlear apex.
hindgut fermentation Fermentative digestion occurring in distal
hematocrit The percentage of total blood volume occupied by red portions of the alimentary canal.
blood cells; in humans, the hematocrit is normally 40 to 50%.
histamine The base formed from histidine by decarboxylation; re-
heme C,,H,,04N4FeOH; an iron protoporphyrin portion of sponsible for dilation of blood vessels.
many respiratory pigments. histaminergic Referring to nerves that release histamine.
hemerythrin An invertebrate respiratory pigment that is a protein histone A simple, repeating, basic protein that combines with
but does not contain heme. DNA.
hemimetabolous Refers to insects, like bugs, that show incomplete Hodgkin cycle The regenerative, or positive-feedback, loop re-
metamorphosis during. their life cycle. See also sponsible for the upstroke of the action potential; depolariz-
holometabolous. ation causes an increase in the sodium permeability,
hemocoel Space between ectoderm and endoderm in many inver- permitting an increased influx of Na+, which further depo-
tebrates containing blood (hemolymph). larizes the membrane.
hemocyanin An invertebrate respiratory pigment that is a protein, holometabolous Refers to insects, like flies, that show complete
contains copper, and is found in mollusks and crustaceans. metamorphosis during their life cycle. See also hemi-
hemoglobin The oxygen-carrying pigment of the erythrocytes, metabolous.
formed by the developing erythrocyte in bone marrow. It is a homeostasis The condition of relative internal stability maintained
complex protein composed of four heme groups and four glo- by physiological control systems.
bin polypeptide chains. They are designated a(alpha),P(beta), homeotherm An animal (mammal, bird) that regulates its own
y(gamma), and S(delta)in an adult, and each is composed of internal temperature within a narrow range, regardless of the
several hundred amino acids. ambient temperature, by controlling heat production and heat
hemolymph The blood of invertebrates with open circulatory sys- loss.
tems. homeoviscous adaptation Molecular level adaptations, especially
hemopoiesis (hematopoiesis) The processes leading to the pro- of cell membranes, that help minimize temperature-induced
duction of blood cells. differences in viscosity.
hemopoietic factor See intrinsic factor. homonymous Pertaining to the same origin.
Henderson-Hasselbalch equation pH = pK + log ([H+accep- homosynaptic modulation A change in synaptic efficacy that
GLOSSARY G-15
...............................................................................
occurs subsequent to activity at a synapse. In synaptic facili- hyperventilation See hyperpnea.
tation, efficacy increases. In synaptic depression, efficacy de- hypoglycemia Low blood glucose levels.
creases. hypoosmotic Containing a lower concentration of osmotically ac-
horizontal cell A nerve cell whose fibers extend horizontally in the tive constituents than the solution of reference.
outer plexiform layer of the vertebrate retina; interconnects hypophysis The pituitary gland.
adjacent photoreceptors. hypopnea Hypoventilation; decreased lung ventilation.
hormone A chemical compound synthesized and secreted by an hypothalamic releasing hormones Hormones from the hypo-
endocrine tissue into the bloodstream; influences the activity thalamus that cause the release of other hormones from the
of a target tissue. pituitary.
horseradish peroxidase A large protein molecule that is opaque in hypothalamus The part of the diencephalon that forms the floor
the electron microscope, and is used to trace neuronal path- of the median ventricle of the brain; includes the optic chi-
ways in the central nervous system. asma, mammillary bodies, tuber cinereum, and infundibulum;
hybridomas Hybrid cells that are formed by the fusion of two dif- many subregions contribute to the regulation of the auto-
ferent cell types, used in the production of monoclonal anti- nomic nervous system and of endocrine function.
bodies and other cellular products. hypothalmo-hypophyseal relay system Portal veins linking the
hydration Combination with water. capillaries of the hypothalamic median eminence with those
hydraulic permeability Refers to the sieve-like properties of the of the adenohypophysis; these transport hypothalamic neu-
Bowman's capsule in the kidney. rosecretions directly to the adenohypophysis.
hydride Any compound consisting of an element or a radical com- hypothermia A state of abnormally low body temperature.
bined with hydrogen. hypothesis A specific prediction that can then be tested by per-
hydrofuge Pertaining to structures with nonwetting surfaces. forming further experiments.
hypothyroidism Reduced thyroid activity.
hydrogen bond A weak electrostatic attraction between a hydro-
gen atom bound to a highly electronegative element in a mol- hypotonic Having a lower tonicity or osmotic pressure than ref-
ecule and another highly electronegative atom in the same or erence solution.
a different molecule. hypoventilation Reduced lung ventilation.
hydrolase transport The mechanism by which monosaccharides hypoxia Reduced oxygen levels.
are taken up into absorptive cells, using a membrane-bound hysteresis A nonlinear change in the physical state of a system,
glycosidase to break down and transport the parent disac- such that the state depends in part on the previous history of
charide across the absorptive cell's membrane. the system.
hydrolysis Fragmentation or splitting of a compound by the addi- I band The region between the A band and Z disk of a resting
tion of water, whereupon the hydroxyl group joins one frag- muscle sarcomere; it appears light when viewed microscopi-
ment and the hydrogen atom the other. cally and contains part of the actin thin filaments that does not
hydronium ion (H,O+)A hydrogen ion (H+)combined with a overlap with myosin filaments.
water molecule; H 2 0 + . ileum Posterior section of the small intestine.
hydrophilic Having an affinity for water. impedance The dynamic resistance to flow met by fluids moving
hydrophobic Lacking an affinity for water. in a pulsatile manner.
hydrostatic pressure Force exerted over an area due to pressure in impulse-initiatingregion (spike-initiatingzone) The proximal por-
a fluid. tion of the axon, which has a lower threshold for action po-
hydroxyapatite Ca,o(PO,),(OH),, a crystalline material lending tential generation than either the soma or the dendrites.
hardness and rigidity to the bones of vertebrates and shells of incus The middle bone of the three bones in the mammalian inner
mollusks. ear; it connects the malleus and the stapes.
hydroxyl group(radica1)The -OH- group. influx Movement of solute or solvent into a cell across the cell
hvdroxvl membrane.
, , ion OH-.
hypercalcemia Excessive plasma calcium levels. incisors Chisel-like teeth used for gnawing.
hypercapnia Increased levels of carbon dioxide. inertia The tendency of a mass to resist acceleration.
infradian rhythms Biological rhythms with a periodicity of less
hyperemia Increased blood flow to a tissue or an organ.
than a day in length.
hyperglycemia Excessive blood glucose levels.
infrared Thermal radiation; electromagnetic radiation of wave-
hyperosmotic Containing a greater concentration of osmotically lengths greater than 7.7 x 10-'cm and less than 12 x 10-4cm;
active constituents than the solution of reference. the region between red light and radio waves.
hyperpnea Increased lung ventilation; hyperventilation. inhibitory In neurophysiology, pertaining to a reduction in prob-
hyperpolarization An increase in potential difference across a ability of generating an action potential.
membrane, making the cell interior more negative than it is at inhibitory postsynaptic potential (ipsp) A change in the trans-
rest. membrane potential of a postsynaptic cell that reduces the
hyperthermia A state of abnormally high body temperature. probability of an action potential in the cell.
hypertonic Having a higher tonicity or osmotic pressure than ref- initial segment The portion of axon and axon hillock proximal
erence solution. to the first myelinated segment; the spike initiating zone of
hypertrophy Excessive growth or development of an organ or a many neurons.
tissue. inner plexiform layer In the vertebrate retina, the layer of
connecting processes that lies between the bipolar cells and iodoacetic acid An agent that poisons glycolysis by inhibiting glyc-
the ganglion cells. eraldehyde phosphate dehydrogenase.
inner segment The portion of a vertebrate photoreceptor cell that ion An atom bearlng a net charge due to loss or gain of electrons.
contains the cell organelles and synaptic contacts. ion battery The electromotive force capable of driving an ionic
inositol trisphosphate (InsP,) The intracellular second messenger current across a membrane; results from unequal concentra-
produced by the action of phospholipase C on membrane tions of an ion species in the two compartments separated by
phosphatidylinositolphosphate in response to stimulation of the membrane.
cell-surface receptors by growth factors, hormones, or neuro- ion bonding sites Partially ionized regions of protein and other
transmitters. molecules that electrostatically interact with ions in the sur-
inotropic Pertaining to the strength of contraction of the heart. rounding solution.
inspiratory neurons Neurons in the medulla of the brain control- ion-exchanger site (ion-binding site) An electrostatically charged
ling motor neurons of muscle associated with breathing in. site that attracts ions of the opposite charge.
instars The stages between molts in insect development. ionic bond Electrostatic bond.
ionization The dissociation into ions of a compound in solution.
instinct A species-specific set of unlearned behaviors and re-
sponses. ipsilateral Relating to the same side.
insulin A protein hormone synthesized and secreted by the beta iris The pigmented circular diaphragm located behind the cornea
cells of the pancreatic islets; controls cellular uptake of car- of the vertebrate eye.
bohydrate and influences lipid and amino acid metabolism. ischemia The absence of blood flow (to an organ or a tissue).
integral proteins Proteins spanning the cell membrane that form islets of Langerhans Microscopic endocrine structures dispersed
selective filters and active transport devices that get nutrients throughout the pancreas. They consist of three cell types: the
into and cellular products and waste out of the cell. alpha cells, which secrete glucagon; the beta cells, which se-
crete insulin; and the delta cells, which secrete gastrin.
integration, neuronal Synthesis of an output based on the sum of
inputs to a neuron or neuronal network. isoelectric point The pH of a solution at which an amphoteric
molecule has a net charge of zero.
intercalated disk The junctional region between two connected
isomer A compound having the same chemical formula as another,
cardiac muscle cells.
but with a different arrangement of its atoms.
interferon anti-viral and anti-tumor agent produced by animal
isometric contraction Contraction during which a muscle does not
cells
shorten significantly.
interneuron A nerve cell that is entirely contained within the cen-
isometry Proportionality of shape regardless of size.
tral nervous system; interneurons typically connect two or
isosmotic Having the same osmotic pressure.
more other neurons.
isoteric interaction Chemical interaction involving molecules with
internode The space along a myelinated axon that is covered by
the same number of valence electrons in the same configura-
the myelinating cell (i.e., it is covered by the Schwann cell or
tion, but made up of different types and numbers of atoms.
the oligodendrocyte).
isotonic Having an equivalent tonicity or osmotic pressure than
interoceptive receptors Internal sensory receptors responding to
reference solution.
changes inside the body.
isotonic contraction Contraction in which the force generated re-
interstitial Between cells or tissues. mains constant while the muscle shortens.
interstitial cell-stimulating hormone (ICSH)Identical to luteiniz- isotope Any of two or more forms of an element with the same
ing hormone but in the male. number of protons (atomic number), but a different number
interstitium The tissue space between cells. of neutrons (atomic weight).
intestinal chyme Semifluid mass of partially digested food. isovolumic Having the same volume.
intestinal gastrin Hormone stimulating gastric glands to increase isozymes Multiple forms of an enzyme found in the same animal
secretion rate. species or even in the same cell.
intestinal juice See succus entericus.
Jacobs-Stewart cycle The cycling of CO, and H C 0 3 between in-
intestinal phase Phase of digestion control by gastrin and other
tracellular and extracellular compartments, which functions
hormones.
to transfer H+ ions between the cell interior and the extracel-
intracellular digestion Nutrient breakdown occurring within cells.
lular fluid.
intracellular milieu The general physio-chemical characteristics jejunum The portion of small intestine between the duodenum and
within the cell. the ileum.
intrafusal fibers The muscle fibers within a muscle spindle organ. joule (J) SI unit of work equivalent to 0.239 calories (cal).
intrinsic factor (or hemopoietic factor) A mucoprotein produced junctional fold A fold in the cell membrane of a postsynaptic cell
by the H+-secreting parietal cells of the stomach; involved in that lies under the axon terminals of the presynaptic cell. Junc-
vitamin BI2 absorption. tional folds are typical of skeletal muscle fibers at neuromus-
inulin An indigestible vegetable starch; used in studies of kidney cular junctions.
function because it is freely filtered and not actively trans- juvenile hormone UH) h class of insect hormones that are secreted
ported. by the corpora allata and that promote retention of juvenile
in vitro "In a glass"; in an artificial environment outside the characteristics.
body. juxtaglomerular apparatus Apparatus comprised of specialized se-
in vivo Wlthm the llv~ngorganlsm or tlssue. cretory cells situated in the afferent glomerular arterioles; act
as receptors that respond to low blood pressure by secreting proportional to the wall tension divided by the inner radius of
renin, which converts angiotensinogen to angiotensin, result- the tube.
ing in vasoconstriction and aldosterone secretion. larva The immature, active feeding stage characteristic of many
juxtaglomerular cells Specialized secretory cells situated in the af- invertebrates.
ferent glomerular arterioles; act as receptors that respond to latent heat of vaporization The amount of energy required to
low blood pressure by secreting renin, which converts an- change a liquid to its gaseous form (evaporate) at the same
giotensinogen to angiotensin, resulting in the stimulation of temperature.
aldosterone secretion. latent period The interval between an action potential in a mus-
juxtapulmonary capillary receptors Sensory receptors found in the cle fiber and the initiation of contraction.
lung that, when stimulated, elicit the sensation of breathless- lateral geniculate A region of the brain in birds and mammals that
ness. processes visual information coming from the retina.
k selection Pattern of energetic investment in reproduction in lateral geniculate nuclei The major relay nuclei between the retina
which small numbers of large offspring are produced, each and the visual cortex in the mammalian visual system; they
with a high chance of survival due to extensive parental care. are included in the thalamic nuclei.
kelvin (K) See absolute temperature. lateral inhibition Reciprocal suppression of excitation by
keratin Structural protein found in skin, feathers, nails, and hoofs. neighboring neurons in a sensory network; it produces
ketone Any compound having a carbonyl group (CO) attached enhanced contrast at boundaries and an increase in dynamic
(by the carbon) to hydrocarbon groups. range.
ketone bodies Acetone, acetoacetic acid, and P-hydroxybutyric lateral-linesystem Series of hair cells (seeneuromast) in canals run-
acid; products of fat and pyruvate metabolism formed from ning the length of the head and body of fishes and many am-
acetyl CoA in the liver; oxidized in muscle and by the central phibians; these channels have openings to the outside, and the
nervous system during starvation. system is sensitive to water movement.
key stimulus (releasing stimulus) The stimulus that is effective in lateral plexus In the compound eye of Limulus, the collection of
producing a fixed action pattern. neurons that interconnect eccentric cells of the ommatidia,
kinematic viscosity Viscosity divided by density; gases of equal producing lateral inhibition.
kinematic viscosity will become turbulent at equal flow rates lecithin Any of a group of phospholipids found in animal and
in identical airways. plant tissues; composed of choline, phosphoric acid, fatty
kinetic energy Energy inherent in the motion of a mass. acids, and glycerol.
kininogen Precursor of bradykinin. length constant (1) The distance along a cell over which a potential
kinocilium A true "9 + 2" or "9 + 0" cilium present in sensory change decays in amplitude by (1- l/e), or 63%.
hair cells. lens The major light-focusing structure in the vertebrate eye.
Kirchhoff's laws First law:The sum of the currents entering a junc- leukocytes White blood cells.
tion in a circuit equals the sum of the currents leaving the Leydig cells (interstitial cells) Cells of the testes that are stimulated
junction. Second law: The sum of the ~otentialchanges en- by luteinizing hormone to secrete testosterone.
countered in any closed loop in a circuit is equal to zero. ligand-gated ion channel An ion channel through the cell mem-
Kleiber's law 0.75 exponent relates metabolic rate to body mass. brane that opens when a molecule, or molecules, binds to the
knock-outs Animals that lack the ability to express the function extracellular domain of the protein.
originally coded for by the gene. light Electromagnetic radiation with wavelengths between those
Krebs cycle See tricarboxylic acid (TCA) cycle and citric acid of x-rays and those of heat (infrared radiation).
cycle. light meromyosin (LMM) The rodlike fragment of the myosin
molecule that constitutes most of the molecule's backbone.
labeled line coding A pattern of information processing in the ner-
vous system in which each neuron encodes only one particu- limited heterothermy A survival mechanism used by camels and
lar type of information (e.g., sour stimuli in the taste system) other normally homeothermic animals, in which body tem-
and all of the axons that carry that type of information pro- perature is allowed to rise and fall somewhat as ambient tem-
ject to the same location or locations. perature ranges through extremes.
labeled-lines concept The idea that sensory modalities are deter- Lineweaver-Burk equation Straight line transformation of the
mined by the stimulus sensitivity of peripheral sense organs Michaelis-Menton equation.
and the anatomical specificities of their central connections. lipases Enzymes that specifically break down lipids.
lactation The production of milk by the mammary glands lipid Any of the fatty acids, neutral fats, waxes, steroids, and phos-
(breasts). phatides; lipids are hydrophobic and feel greasy.
lactogen Hormone that prepares the breasts for milk production. lipid bilayer Continuous double layer of lipid molecules forming
lagena A structure associated with hair cells in the vertebrate the basic structure of the cell membrane.
organs of equilibrium. lipogenesis The formation of fat from nonlipid sources.
lamella A thin sheet or leaf. lipophilic Having an affinity for lipids.
laminar flow Turbulence-free flow of fluid in a vessel or past a lipoprotein Protein-lipid complex in the plasma membrane.
moving object; a gradient of relative velocity exists in which local circuit current The current that spreads electrotonicallyfrom
the fluid layers closest to the wall or body have the lowest rel- the excited portion of an axon during conduction of the nerve
ative velocity. impulse, flowing longitudinally along the axon, across the
Laplace's law The transmural pressure in a thin-walled tube is membrane, and back to the excited portion.
G-18 GLOSSARY
......................................
longitudinal smooth muscle Outer layer of smooth muscle running mechanism A theory that proposes that life is based purely on the
along the long axis of small intestine. action of physical and chemical laws.
long-term potentiation An increase in synaptic efficacy that occurs mechanoreceptor A sensory receptor tuned to respond to me-
due to sustained synaptic input and that lasts for a relatively chanical distortion or pressure.
long time-even days, weeks, or months. median eminence A structure at the base of the hypothalamus
loop of Henle A U-shaped bend in the portion of a renal tubule that is continuous with the hypophyseal stalk; contains the
that lies in the renal medulla. primary capillary plexus of the hypothalamo-h~pophyseal
lower critical temperature (LCT) ambient temperature below portal system.
which the BMR becomes insufficient to balance heat loss, re- medulla oblongata In vertebrates, a cone-shaped neuronal mass
sulting in falling body temperature. that lies between the pons and the spinal cord.
lumen The interior of a cavity or duct. medullary cardiovascular center A group of neurons in the
luminosity Brightness; relative quantity of light reflected or medulla involved in the integration of information used in the
emitted. control and regulation of circulation.
luteal phase The part of the estrous or menstrual cycle character- medullary respiratory centers Groups of neurons in the medulla of
ized by formation of and secretion by the corpus luteum. the brain controlling the activity in motor neurons associated
luteinizing hormone (LH) A gonadotropin that is secreted by the with breathing.
aden~h~pophysis and that acts with follicle-stimulating hor- melanocyte-stimulating hormone (melanophore-stimulatinghor-
mone (FSH)to induce ovulation of the ripe ovum and libera- mone) A peptide hormone released by the adenohypophysis
tion of estrogen from the ovary; also influences formation of that effects melanin distribution in mammals and creates skin-
the corpus luteum and stimulates growth in and secretion color changes in fishes, amphibians, and reptiles.
from the male testicular Leydig cells. melting point The lowest temperature at which a solid will begin
lymph Plasmalike fluid collected from interstitial fluid and re- to liquefy.
turned to the bloodstream via the thoracic duct; contains membrane potential The electric potential measured from within
white, but not red, blood cells. the cell relative to the potential of the extracellular fluid,
lymphatic system A collection of blind-ending tubes which drain which is by convention at zero potential; the potential differ-
filtered extracellular fluid from tissues and return it to the ence between opposite sides of the membrane.
blood circulation. membrane pumps Membrane-based cellular mechanisms that ac-
lymph heart A muscular pump found in fish and amphibia caus- tively transport substances against a gradient.
ing movement of lymph.
membrane recycling Recovery and reformation into new secretory
lymph nodes Aggregations of lymphoid tissue in the lym- vesicles of membrane lost from the cell membrane due to
phatic system that produce lymphocytes and filter the lym- exocytosis.
phatic fluid.
membrane transport proteins Integral proteins that transport par-
lymphocytesWhite blood cells, produced in lymphoid tissue, lack- ticular classes of molecules across membranes; see active
ing cytoplasmic granules but having a large, round nucleus. transport.
lysolecithin A lecithin without the terminal acid group.
menarche The onset of menstruation during puberty.
lysosomes Minute electron-opaque organelles that occur in many
menopause The cessation of the menstrual cycle in the mature fe-
cell types, contain hydrolytic enzymes, and are normally in-
male human.
volved in localized intracellular digestion.
menses Shedding of the uterine lining during a menstrual cycle.
M line In a muscle sarcomere, the darkly staining structure in the menstrual cycle Recurring physiological changes that include men-
middle of the H band. struation.
maculae (Greek for "spots.") Organs of equilibrium in the verte- menstruation The shedding of the endometrium, an event
brate inner ear. that usually occurs in the absence of conception throughout
magnetite A magnetic mineral composed of Fe,O, and found the fertile period of the female of certain primate species, in-
in some animals; believed to play a role in geomagnetic cluding humans.
orientation. mesencephalicus lateralis dorsalis (MLD)The nucleus to which
malleus The outermost bone of the three bones in the mammalian auditory information projects in the owl, contributing to the
inner ear; it connects the tympanic membrane with the incus. bird's ability to locate objects in its environment based entirely
Malpighian tubules Insect excretory osmoregulatory organs on audition.
responsible for the active secretion of waste products and the messenger molecules Hormones, synaptic transmitters, and other
formation of urine. chemicals that regulate biological processes.
mass action law The velocity of a chemical reaction is propor- messenger RNA (mRNA)A fraction of RNA that is responsible
tional to the active masses of the reactants. for transmission of the informational base sequence of the
mass-specific metabolic rate The metabolic rate of a unit mass of DNA to the ribosomes.
tissue. metabolic acidosis A decrease in blood pH at constant PCO, usu-
mastication The chewing or grinding of food with the teeth. ally as a result of metabolism or kidney function.
mastoid bone The posterior process of the temporal bone, situated metabolic alkalosis An increase in blood pH at constant PCO,
behind the ear and in front of the occipital bone. usually as a result of metabolism or kidney function.
maximum aerobic velocity (MAV)Locomotor speed at which the metabolic intensity The metabolic rate of a unit mass of tissue. See
maximum rate of aerobic respiration is reached. also mass-specific metabolic rate.
GLOSSARY G-19
. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .' .
metabolic pathway A sequence of enzymatic reactions involved in mobility, mechanical A quantity proportional to the rate at which
the alteration of one substance into another. a molecule will diffuse in a liquid phase.
metabolic water Water evolved from cellular oxidation. modulatory agent One that either increases or decreases the re-
metabolism The totality of physical and chemical processes in- sponse of a tissue to a physical or chemical signal.
volved in anabolism, catabolism, and cell energetics. molality The number of moles of solute in a kilogram of a pure
metachronal waves Waves of activity that spread over a popula- solvent.
tion of beating cilia. molarity The number of moles of solute in a liter of solution.
metachronism The progression of in-phase activity in a wavelike molars Teeth used in a side-by-side grinding motion to break
manner over a population of organelles, such as cilia. down food.
metamorphic climax The last stage of amphibian metamorphosis, mole Avogadro's number (6.023 x loz3)of molecules of an ele-
in which the adult form is attained. ment or a compound; equal to the molecular weight in grams.
metamorphosis A change in morphology-in particulal; from one
molecular phylogeny A system of phylogenetic relations inferred
stage of development to another, such as juvenile to adult.
from similarities and differences in the nucleic acid sequences
metarhodopsin Product of the absorption of light by rhodopsin; coding for identified proteins.
decomposes to opsin and trans-retinal.
monoclonal antibody A homogeneous antibody that is produced
metarteriole An arterial capillary.
by a clone of antibody-forming cells and that binds with a sin-
metazoa Multicellular organisms. gle type of antigen.
methemoglobin Hemoglobin in which the Fe3+of heme has been monocytes White blood cells lacking cytoplasmic granules, but
oxidized to Fez+.
having an indented or horseshoe-shaped nucleus.
micelle A microscopic particle made from an aggregation of
monogastric stomach A stomach consisting of a single strong mus-
amphipathic molecules in solution.
cular tube or sac.
Michaelis-Menton equation The rate equation for a single
monomer A compound capable of combining in repeating units to
enzyme- catalyzed substrate reaction.
form a dimer, trimer, or polymer.
miaoclimate A small refugium (e.g., burrow, crack in bark) that
monopole An object or a particle bearing a single unneutralized
provides protection from general climatological conditions.
electric charge, as, for example, an ion.
microelectrodes Tiny glass "needles" inserted into tissues or even
individual cells for recording physiological data. monosaccharide sugar An unhydrolyzable carbohydrate, a simple
microfilaments Actin filaments within the cytoplasmic substance; sugar. Such sugars are sweet-tasting, colorless crystalline com-
diameter of less than 10 nm. pounds with the formula C,(H20), . See also saccharide.
micromanipulator A mechanical device that holds and moves mi- monosynaptic Requiring or transmitted through only one synapse;
croelectrodes incrementally in three different planes. for example, the stretch reflex of vertebrate limbs.
miaotome A device used in microscopy to cut ultrathin sections monovalent Having a valence of one.
from small blocks of tissues. monozygotic Arising from one ovum or zygote.
microtubules Cylindrical cytoplasmic structures made of motility The ability of the alimentary tract to contract and trans-
polymerized tubulin and found in many cells, especially port ingested material along its length.
motile cells, as constituents of the mitotic spindle, cilia, and motor cortex The part of the cerebral cortex that controls motor
flagella. function; situated anterior to the central sulcus, which sepa-
microvilli Tiny cylindricalprojections on a cell surface that greatly rates the frontal and parietal lobes.
increase surface area; frequently found on absorptive motor neuron (motoneuron) A nerve cell that innervates muscle
epithelia, but also in photoreceptors. fibers.
micturition Urination. motor program An endogenous coordinated motor output of cen-
midgut Major alimentary canal site for the chemical digestion of tral neuronal origin and independent of sensory feedback.
protein, fat, and carbohydrates. motor unit The unit of motor activity consisting of a motor neu-
mineralocorticoids Steroid hormones that are synthesized and se- ron and the muscle fibers it innervates.
creted by the adrenal cortex and that influence plasma elec- mRNA See messenger RNA.
trolyte balance-in particular, by sodium and chloride
mucin The mucopolysaccharide forming the chief lubricant of
reabsorption in the kidney tubules. See also aldosterone.
mucus.
miniature endplate potentials (mepps) Tiny depolarizations (gen-
mucosa Mucous membrane facing a cavity or the exterior of the
erally 1 mV or less) of the postsynaptic membrane at a mo-
body.
tor endplate; produced by presynaptic release of single packets
of transmitter. mucosal See mucosa.
miniature postsynaptic potentials (mpps)Potentials produced in a mucous cells Mucus-secreting cells of the intestine.
postsynaptic neuron by presynaptic release of single vesicles mucus A viscous, protein-containing mixture of mucopolysac-
of transmitter substance. charides secreted from specialized mucous membranes; often
mitochondria Membrane-enclosed organelle where ATP is pro- plays an important role in filter feeding (invertebrates)or in
duced during aerobic metabolism. lubricating or protecting internal or external surfaces.
mixed nerve A nerve that contains axons of both sensory and mo- Mullerian ducts Paired embryonic ducts originating from the peri-
tor neurons. toneum that connect with the urogenital sinus to develop into
mobility, electrical A quantlty proportional to the migration rate the uterus and fallopian tubes.
of an ion in an electric field. multineuronal innervation Innervation of a muscle fiber by
G-20 GLOSSARY
.........................................
several motor neurons, as in many invertebrates, especially pothesis that the sensory modality of a stimulus is encoded in
arthropods. the projection pattern of sensory neurons and not on partic-
multiterminal innervation Numerous synapses made by a single ular features of the cellular response in the stimulated
motor neuron along the length of a muscle fiber. neurons.
multi-unit muscle A smooth muscle in which individual muscle nervous system The collection of all neurons in an animal's body.
fibers contract only when they receive excitatory input from net flux Sum of influx and efflux through a membrane or other
neurons; contraction of these muscles is neurogenic. material.
muscarinic Pertaining to muscarine, a toxin derived from mush- neurilemma Connective tissue sheath covering a bundle of axons.
rooms; refers to acetylcholine receptors that respond to mus- neurites Cell processes extending from the soma of neurons.
carine, but not to nicotine. neurogenic pacemaker A pacemaker that is a specializednerve cell.
muscle fiber A skeletal muscle cell. neuroglia (glia) Inexcitable supporting tissue of the nervous
muscle spindle (stretch receptor) A length-sensitive receptor or- system.
gan located between and in parallel with extrafusal neurohemal organ Organ for storage and discharge into the blood
muscle fibers; gives rise to the myotatic, or stretch, reflex of of the products of neurosecretion.
vertebrates. neurohormone A substance that exists within the neurons of the
mutagens Compounds that produce mutations in the germ cell nervous system and exert hormonal effects outside the ner-
line. vous system.
mutation A transmissible alteration in genetic material. neurohumor Synaptic transmitters and neurosecretory hormones.
myelination Forming a myelin sheath. neurohypophysis (pars nervosa) A neuronally derived reservoir
myelin sheath A sheath formed by many layers of the membrane for hormones with antidiuretic and oxytocic action; consists
of Schwann cells or oligodendrocyte glial cells that are of the neural lobe, which makes up its bulk, and the neural
wrapped tightly around segments of axon in vertebrate nerve; stalk, which is connected to and passes neurosecretions from
serves as electrical insulation in saltatory conduction. the hypothalamus.
myoblast Embryonic precursor for skeletal muscle fibers. neuromast A collection of hair cells embedded in a cupula in
myocardium Heart muscle. lateral-line mechanoreceptor of the lower vertebrates.
myofibril A longitudinal unit of muscle fiber made up of sarcom- neuromodulation A change in neuronal function caused by chem-
eres and surrounded by sarcoplasmic reticulum. ical messengers (neuromodulators) that are released from
myogenic Capable of producing an intrinsic cycle of electrical ac- axon terminals, but that diffuse more widely than do typical
tivity. neurotransmitters; neuromodulatory effects can be relatively
myogenic pacemaker A pacemaker that is a specialized muscle cell. long-lasting.
myoglobin An iron-containing protoporphyrin-globin complex neuromuscular junction (NMJ)The synapse that connects a mo-
found in muscle; serves as a reservoir for oxygen and gives tor neuron with a skeletal muscle fiber.
some muscles their red or pink color. neuron Nerve cell.
myoplasm The cytosol in a muscle cell. neuronal circuit A set of interconnected neurons.
myosin The protein that makes up the thick filaments and cross neuronal integration Ongoing summing of all synaptic input onto
bridges in muscle fibers; it is also found in many other cell a postsynaptic cell, which determines whether or not the post-
types and is associated with cellular motility. synaptic cell will produce an action potential.
myotatic reflex (stretch reflex) Reflex contraction of a muscle in re- neuronal network (neuronal circuit) A system of interacting nerve
sponse to stretch of the muscle. cells.
myotube A developing muscle fiber. neuronal plasticity Modification of activity in a neuronal circuit
based on experience and changes in input.
Na+-K+ pump See sodium-potassium pump. neuropeptide A peptide molecule identified as a neurotransmitter
Naloxone An analog of morphine that acts as an opioid substance.
antagonist. neuropeptide Y A 36 amino acid peptide, co-localized with nor-
nares Nostrils. epinephrine in sympathetic ganglia and adrenergic nerves as
nematocysts Stinging cells of hydras, jellyfish, and anemones. well as localized in some nonadrenergic fibers, the physiolog-
nephron The morphological and functional unit of the vertebrate ical effects of which include amelioration of actions of cate-
kidney; composed of the glomerulus and Bowman's capsule, cholamines on the mammalian heart and potentiation of
the proximal and distal tubules, the loop of Henle (birds, actions of catecholamines in fish hearts.
mammals), and the collecting duct. neurophysins Proteins associated with neurohypophyseal hor-
Nernst equation Equation for calculating the electrical potential mones stored in granules in the neurosecretory terminals;
difference across a membrane that will just balance the con- cleaved from the hormones before secretion.
centration gradient of an ion. neuropil A dense mass of closely interwoven and synapsing
nerve As a noun: A bundle of axons held together as a unit by con- nerve cell processes (axon collaterals and dendrites) and glial
nective tissue. As an adjecttve: Neuronal. cells.
nerve net A collection of interconnected neurons that are distrib- neurosecretory cells Nerve cells that liberate neurohormones.
uted through the body, rather than concentrated in a central neurotoxin A substance that interferes with the proper firing of
location; these neuronal systems are most typical of lower or- nerve impulses.
ganisms, such as coelenterates. neurotransmitter A chemical mediator released by a presynaptic
nerve-specific energy The term used by Johannes Muller in his hy- nerve ending that interacts with receptor molecules in the
postsynaptic membrane. This process generally induces a tance of a column of mercury 1 mm2 in cross-sectional and
permeability increase to an ion or ions and thereby influences 106 cm long.
the electrical activity of the postsynaptic cell. Ohm's law I = V I R. The strength of an electric current, I, varies
nicotinamide adenine dinucleotide (NAD)A coenzyme widely dis- directly as the voltage, V, and inversely as the resistance, R.
tributed in living organisms, participating in many enzymatic olfaction The sense of smell; chemoreception of molecules sus-
reactions; made up of adenine, nicotinamide, and two mole- pended in air.
cules each of d-ribose and phosphoric acid. oligodendrocytes A class of glial cells with few processes. These
nicotinic Pertaining to nicotine, an alkaloid derived from tobacco; cells wrap axons in the central nervous system, forming
refers to acetylcholine receptors that respond to nicotine, but myelin sheaths.
not to muscarine. oligopeptides Polypeptide residues of two or three amino acids.
node of Ranvier Regularly spaced interruption (about every mil- oligosaccharides Carbohydrates made up of a small number of
limeter) of the myelin sheath along an axon. monosaccharide residues.
noncompetitive inhibition Enzyme inhibition due to alteration or omasum The part of the ruminant stomach lying between the ru-
destruction of the active site. men and the abomasum.
nonsaturation kinetics Kinetics occurring when the rate of influx ommatidium The functional unit of the invertebrate compound
increases in proportion to the concentration of the solute in eye, consisting of an elongated structure with a lens, a focus-
the extracellular fluid. ing cone, and photoreceptor cells.
nonshivering thermogenesis A thermogenic process in which en- oncotic pressure Osmotic pressure plus hydrostatic pressure
zyme systems for fat metabolism are activated, breaking down caused by distribution of ions according to the Donnan equi-
and oxidizing conventional fats to produce heat. librium.
nonspiking neuron A neuron that receives and transmits informa- l a afferent fiber An axon with a peripheral sensory ending inner-
tion without action potentials; synaptic transmitters are re- vating a muscle spindle organ and responding to stretch of the
leased by these neurons in proportion to their membrane organ; its central terminals synapse directly onto alpha motor
potential, a process called nonspiking release. neurons of the homonymous muscle.
nonspiking release The release of neurotransmitter from a pre- l b afferent fiber An axon whose sensory terminals innervate the
synaptic neuron that occurs independent of action potentials; tendons of skeletal muscle and respond to tension.
typically graded changes in membrane potential modulate the 1,25-dihydrozycholecalciferolA substance that is converted from
activation of voltage-gated Ca2+channels, and changes in the vitamin D in the liver and increases Ca2+absorption by the
intracellular concentration of free Ca2+modulate the release kidney.
of transmitter. onophores Molecules or molecular aggregates that promote the
norepinephrine (noradrenaline) A neurohumor secreted by the pe- permeation of ions across membranes; these may be carrier
ripheral sympathetic nerve terminals, some cells of the central molecules or ion-permeable membrane channels.
nervous system, and the adrenal medulla. oocyte A developing ovum.
nucleases Enzymes that hydrolyze nucleic acids and their residues. operator gene A gene that regulates the synthetic activity of closely
nucleic acids Nucleotide polymers of high molecular weight. See linked structural genes via its association with a regulator
also DNA; RNA. gene.
nucleosidases Enzymes that hydrolyze nucleic acids and their operon A segment of DNA consisting of an operator gene and its
residues. associated structural genes.
nucleotide A product of enzymatic (nuclease) splitting of nucleic opiates Opium-derived narcotic substances.
acids; made up of a purine or pyrimidine base, a ribose or de- opioids Substances that exert opiatelike effects; some are synthe-
oxyribose sugar, and a phosphate group. sized endogenously by neurons within the vertebrate central
nucleus Of an atom: The central, positively charged mass sur- nervous system.
rounded by a cloud of electrons. Of a cell: The membrane- opsin Protein moiety of visual pigments; it combines with
bound body within eukaryotic cells that houses the genetic 11-cisretinal to become a visual pigment.
material of the cell. Of newe cells: A related group of neurons optic axis An imaginary straight line passing through the center of
in the central nervous system. curvature of a simple lens.
nymph A juvenile developmental stage in some arthropods; mor- optic chiasm (optic chiasma) A swelling under the hypothalamus
phology resembles the adult. of the vertebrate brain where the two optic nerves meet; de-
nystatin A rod-shaped, antibiotic molecule that creates channels pending on the species, some axons cross the midline here and
through membranes that allow the passage of molecules of a project to the contralateral side of the brain.
diameter less than 0.4 nm. optic tectum Region of brain in fish and amphibia involved in
obligatory osmotic exchange An exchange between an animal and processing visual information coming from the retina.
its environment that is determined by physical factors beyond organ of Corti The tissue in the cochlea of the inner car that con-
the animal's control. tains the hair cells.
occipital lobe The most posterior region of the cerebral organs of equilibrium Regions of the inner ear that sense the
hemsphere. position of the body relative to gravity or changes of the
occular dominance column A set of neurons arranged vertically body's position with respect to gravity.
through the mammalian visual cortex, all of which receive in- ornithine-urea cycle A cyclic succession of reactions that elimi-
put from one of the two eyes. nate ammonia and produce urea in the liver of ureotelic
ohm (a) MKS unit of electrical resistance, equivalent to the resis- organisms.
G-22 GLOSSARY
.........................................
osmoconformer An organism that exhibits little or no osmoregu- between the extent of combination of oxygen with the respi-
lation, so that the osmolarity of its body fluids follows ratory pigment and the partial pressure of oxygen in the gas
changes in the osmolarity of the environment. phase.
osmolarity The effective osmotic pressure. oxyhemoglobin Hemoglobin with oxygen combined to the Fe
osmole The standard unit of osmotic pressure. atom of the heme group.
osmolyte A substance that serves the special purpose of raising oxyntic cells (parietal cells) HCl-secreting cells of the stomach
the osmotic pressure or lowering the freezing point of a body lining.
fluid. oxyregulator Animal that maintains oxygen consumption as am-
osmometer An instrument for the measurement of the osmotic bient oxygen falls.
pressure of a solution. oxytocin An octapeptide hormone secreted by the neurohypoph-
osmoregulation Maintenance of internal osmolarity with respect ysis; stimulates contractions of the uterus in childb~rthand the
to the environment. release of milk from mammary glands.
osmoregulator An organism that controls its internal osmolarity P-wave That portion of the electrocardiogram associated with de-
in the face of changes in environmental osmolarity. polarization of the atria.
osmosis The movement of pure solvent from a solution of an area pacemaker An excitable cell or tissue that fires spontaneously and
of low solvent concentration to an area of high solvent con- rhythmically.
centration through a semipermeable membrane separating the pacemaker potentials Spontaneous and rhythmical depolariza-
two solutions. tions produced by pacemaker tissue.
osmotic flow The solvent flux due to osmotic pressure. Pacinian corpuscles Pressure receptors found in skin, muscle,
osmotic pressure Pressure that can potentially be created by os- joints, and connective tissue of vertebrates; they consist of a
mosis between two solutions separated by a semipermeable nerve ending surrounded by a laminated capsule of connec-
membrane; the amount of pressure necessary to prevent os- tive tissue.
motic flow between the two solutions. pancreas An organ that produces exocrine secretions, such as di-
ossicles Little bones. Auditory ossicles are the tiny bones (malleus, gestive enzymes, as well as endocrine secretions, including the
incus, stapes) of the middle eal; which transmit sound vibra- hormones insulin and glucagon.
tions from the tympanic membrane to the oval window. pancreatic duct Duct carrying secretions from the pancreas to the
ostia The small, mouth-like openings in the body wall of sponges. small intestine.
otolith A calcareous particle that lies on hair cells in the organs of pancreatic juice A secretion of the pancreas containing proteases,
equilibrium. li~ases,and carboh~drasesessential for intestinal digestion.
ouabain Cardiac glycoside, a drug capable of blocking some pancreozymin See cholecystokinin.
sodium pumps. parabiosis The experimental connection of two individuals to al-
outer plexiform layer In the vertebrate retina, the layer of con- low mixing of their body fluids.
necting processes that lies between the photoreceptor cells and parabronchi Air-conduction pathways in the bird lung.
the bipolar cells. paracellular pathways Solvent and solute pathway through
outer segment The part of a vertebrate photoreceptor that con- epithelium passing between rather than through cells.
tains the pigmented receptor membranes; it is attached to the paracrine A hormonal pathway characterized by the production
inner segment by a thin bridge that has microfilaments of a biologically active substance that passes by diffusion
arranged as in a cilium. within the extracellular space to a nearby cell where it initiates
oval window The connection between the middle ear and the a response.
cochlea; it is covered by the base of the stapes. parafollicular cells (C cells) Cells in the mammalian thyroid that
overshoot The reversal of membrane potential during an action secrete calcitonin.
potential; the voltage above zero to the peak of the action parallel processing A pattern of information processing in the ner-
potential. vous system in which multiple pathways simultaneouslycarry
ovulation The release of an ovum from the ovarian follicle. information about a particular input or output; the informa-
ovum An egg cell; the reproductive cell (gamete) of the female. tion carried in multiple channels is synthesized where the
oxidant An electron acceptor in a reaction involving oxidation and pathways converge.
reduction. parasympathetic nervous system The craniosacral part of the au-
oxidation Loss of electrons or increase in net positivity of an atom tonomic nervous system; in general, increased activity of these
or a molecule. Biological oxidations are usually achieved by neurons supports vegetative functions such as digestion.
removal of a pair of hydrogen atoms from a molecule. parathyroid glands Small tissue masses (usuallytwo pairs) close to
oxidative phosphorylation Respiratory chain phosphorylation; the the thyroid gland that secrete parathormone (parathyroid
formation of high-energy phosphate bonds via phosphoryla- hormone).
tion of ADP to ATP, accompanied by the transport of elec- parathyroid hormone (PTH; parathormone) A polypeptide hor-
trons to oxygen from the substrate. mone of the parathyroid glands secreted in response to a low
oxyconformer Animal that allows oxygen consumption to fall as plasma calcium level; stimulates calcium release from bone
ambient oxygen falls. and calcium absorption by the intestines while reducing cal-
oxygen debt The extra oxygen necessary to oxidize the products cium excretion by the kidneys.
of anaerobic metabolism that accumulate in the muscle tissues paraventricular nucleus A group of neurosecretory neurons in
during intense physical activity. the supraoptic hypothalamus that send their axons into the
oxygen dissociation curves Curves that describe the relationship neurohypophysis.
GLOSSARY G-23
......................................
parietal cells Cells of the stomach lining that secrete hydrochloric ment for the purpose of signaling between individuals of the
acid. See also oxyntic cells. same species.
pars intercerebralis The dorsal part of the insect brain; contains phlorizin A glycoside that inhibits active transport of glucose.
the cell bodies of neurosecretory cells that secrete brain hor- phonon A quantum of sound energy.
mones from axon terminals in the corpora cardiaca. phosphagens High-energy phosphate compounds (e.g., phospho-
partition coefficient Ratio of the distribution of a substance be- arginine and phosphocreatine) that serve as phosphate-group
tween two different liquid phases (e.g., oil and water). donors for rapid rephosphorylation of ADP to ATP.
parturition The process of giving birth. phosphoarginine A compound that has phosphagen properties
pawalbumin Calcium-binding protein found in vertebrate similar to those of phosphocreatine and that occurs in the
muscle. muscles of some invertebrates.
patch-clamping A recording technique in which a glass pipette phosphocreatine (aeatine phosphate) A phosphorylated nitroge-
electrode is brought into contact with the outside of a cell nous compound found primarily in muscle; contains a high-
membrane and sealed tightly against it. Current is then passed energy phosphate bond, which can be rapidly transferred to
to hold the potential difference across the membrane at a con- ADP, regenerating ATP.
stant value, and the magnitude of the current required is phosphodiesterase An hydrolytic cytoplasmic enzyme that de-
recorded. The method can be used to measure ionic currents grades CAMPto AMP.
through single ion channels or across the membrane of an en- phosphodiester bonds Bonds that link individual nucleotides in
tire cell. nucleic acids.
patch clamp recording A method of investigating epithelial ion phosphodiester group -0 -P-0 -.
current transfer on very localized regions of cells. phosphoglycerides Glycerine-based lipids of cell membrane.
patella The bone of the knee-cap. phospholipid A phosphorus-containing lipid that hydrolyzes to
pentose A five-carbon monosaccharide sugar. fatty acids, glycerin, and a nitrogenous compound.
pepsin A proteolytic enzyme secreted by the stomach lining. phosphorylase a Activated (phosphorylated)form of phosphory-
pepsinogen Proenzyme of pepsin. lase that catalyzes the cleavage of glycogen to glucose 1-
peptide A molecule consisting of a linear array of amino residues. phosphate.
Protein molecules are made of one or more peptides. Short phosphorylase kinase Enzyme that, when phosphorylated by a
chains are oligopeptides; long chains are polypeptides. protein kinase, converts phosphorylase b to the more active
phosphorylase a.
peptide bond The center bond of the -CO-NH- group, cre-
ated by the condensation of amino acids into peptides. phosphorylation The incorporation of a PO3- group into an or-
ganic molecule.
peptide hormones Hormones that regulate alimentary canal basic
photon A quantum of light energy (the smallest amount of light
electric rhythms.
that can exist at each wavelength).
perfusion The passage of fluid over or through an organ, a tissue,
photopigments Pigment molecules that change their energy state
or a cell.
when they absorb one or more photons of light.
pericardium The connective-tissue sac that encloses the heart.
photoreceptor A sensory cell that is tuned to receive light energy.
perilymph The aqueous solution, similar to other body fluids, that
physiological dead space That portion of inhaled air not involved
is contained within the scala tympani and scala uestibuli of the
in gas transfer in the lung.
cochlea.
pilomotor Pertaining to the autonomic control of smooth muscle
peripheral nervous system The set of neurons and parts of neurons
for the erection of body hair.
that lie outside of the central nervous system.
pinna The outer structure of the mammalian ear, which can be
peripheral resistance units (PRUs) The drop in pressure (in mil- more or less elaborate and which captures and funnels sound
limeters of mercury, mmHg) along a vascular bed divided by
into the ear.
mean flow in milliliters per second.
pinnate Resembling a feather, with similar parts arranged on op-
peristalsis A traveling wave of constriction in tubular tissue pro- posite sides of the axis.
duced by contraction of circular muscle.
pinocytosis Fluid intake by cells via surface invaginations that seal
peritoneum The membrane that lines the abdominal and pelvic off to become vacuoles filled with liquid.
cavities.
pituitary gland (hypophysis) A complex endocrine organ situated
permeability The ease with which substances can pass through a at the base of the brain and connected to the hypothalamus by
membrane. a stalk. It is of dual origin: the anterior lobe (adenohypoph-
pH scale Negative log scale (base 10) of hydrogen ion concentra- ysis) is derived from embryonic buccal epithelium, whereas
tion of a solution. pH = - log[H+]. the posterior lobe is derived from the diencephalon.
phagocyte A cell that engulfs other cells, microorganisms, or for- pK' The negative log (base 10) of an ionization constant,
eign particulate matter. K' .pk' = -log,,K1.
phagocytosis The ingestion of particles, cells, or microorganisms placebo A physiologically neutral substance that elicits curative or
by a cell into its cytoplasmic vacuoles. analgesic effects, apparently through psychological means.
phase contrast microscopy A microscopic technique using differ- placental lactogen A hormone from the placenta that prepares the
ential light refraction by different components of the specimen breasts for milk production.
to enhance viewed images. plane-polarized light Light vibrating in only one plane.
phasic Transient. plasma kinins Peptide hormones formed in the blood after in-
pheromone A species-specificsubstance released into the environ- jury-for example, bradykinin.
G-24 GLOSSARY
......................................
plasmalemma Cell membrane; surface membrane. potassium activation An increase in the conductance of a mem-
plasma membrane Cell membrane; surface membrane. brane to potassium in response to depolarization.
plasma skimming The separation of plasma from blood w i h the potential The voltage above zero to the peak of the action
circulation. potential.
plasticity Compliance to external influence. potential energy Stored energy that can be released to do work.
plastron The ventral shell of a tortoise or turtle; also a gas film held premetamorphosis The developmental stage just preceding am-
in place under water by hydrofuge hairs, creating a large air- phibian metamorphosis, during which iodine binding and
water interface. hormone synthesis occur in the thyroid gland.
pleura The membranes that line the pleural cavity. presbyopia The tendency for human eyes to become less able to
pleural cavity The cavity between the lungs and the wall of the focus on close objects ("far-sighted") with age; occurs as the
thorax. lens becomes less compliant.
plicae circularis The extensive fold of intestinal mucosa bearing pressure pulse The difference between the systolic and diastolic
villi. pressures.
pneumotaxic center A group of neurons in the pons, thought presynaptic Located on the sending side of a synaptic connection.
to be involved in the maintenance of rhythmic breathing in presynaptic inhibition Neuronal inhibition resulting from the ac-
mammals. tion of a terminal that ends on the presynaptic terminal of an
pneumothorax Collapse of the lung due to a puncture into the excitatory synapse, reducing the amount of transmitter re-
pleural cavity of the chest wall or the lung. leased.
poikilotherm An animal whose body temperature tends to fluctu- primary follicle An immature ovarian follicle.
ate more or less with the ambient temperature. primary projection cortex A region of cerebral cortex that
Poiseuille's law In laminar flow, the flow is directly propor- directly receives sensory signals from lower centers; the first
tional to the driving pressure, and resistance is independent of cortical cells to receive sensory information projected to the
flow. brain.
Poisson distribution A theoretical description of the probability primary sensory neurons Neurons that directly receive sensory
that random, independent events based on a unitary event of stimulation.
a particular size will occur. primary structure The sequence of amino acid residues of a
polyclonal Derived from different cell lines or clones. polypeptide chain.
polyestrous The state of having many estrous cycles throughout proboscis An elongated, protrudmg mouth part, typically in suck-
the year. ing insects.
polymer A compound composed of a linear sequence of simple procaine 2-diethylaminoethyl-p-aminobenzoate; a local anesthetic
molecules or residues. that interferes with some of the ion conductances of excitable
polypeptide chain A linear arrangement of more than two amino membranes.
acid residues. proenzyme (zymogen) The inactive form of an enzyme before it
polypnea Increased breathing rate. is activated by removal of a terminal segment of peptide.
polysaccharides Carbohydrases that hydrolyze the glycosidic progesterone A hormone of the corpus luteum, adrenal cortex,
bonds of long-chain carbohydrates (cellulose,glycogen, and and the placenta that promotes growth of a suitable uterine
starch). lining for implantation and development of the fertilized
polysynaptic Referring to transmission through multiple synapses ovum.
in series. prolactin An adenohypophyseal hormone that stimulates milk
polytene Having many duplicate chromatin strands. production and lactation after parturition in mammals.
pons The region of the vertebrate brain that lies just rostra1 to the prometamorphosis The first stage of amphibian metamorphosis,
medulla oblongata. during which there is increased development and activity in
pores of Kohn Small holes between adjacent regions of the lung, the thyroid gland and median eminence.
permitting collateral air flow. propeptide A large peptide that contains the amino acid sequences
porphyrins A group of cyclic tetrapyrrole derivatives. of several smaller peptides, which are released when the large
porphyropsin A purple photopigment, based on ll-cis dehy- peptide is enzymatically cleaved.
droretinal, that is present in the retinal rods of some freshwa- proprioceptors Sensory receptors situated primarily in muscles
ter fishes. and tendons that relay information about the position and
portal vessels Blood vessels that carry blood directly from one cap- motion of the body.
illary bed to another. prostaglandins A family of natural fatty acids that arise in a vari-
positive phototaxis Referring to the movement of an animal to- ety of tissues and are able to induce contraction in uterine and
ward light. other smooth muscle, lower blood pressure, and modify the
postsynaptic Located on the receiving side of a synaptic actions of some hormones.
connection. prostate gland A gland located around the neck of the bladder and
posttetanic depression Reduced postsynaptic response following urethra in males that contributes to the seminal fluid.
prolonged presynaptic stimulation at a high frequency; be- prosthetic group An organic compound essential to the function
lieved to be due to presynaptic depletion of transmitter. of an enzyme. Prosthetic groups differ from coenzymes in
posttetanic potentiation (PTP)Increased efficacy of synaptic trans- that they are more firmly attached to the enzyme protein.
mission following presynaptic stimulation at a high frequency; protagonistic muscles Muscles whose contractions cooperate to
often follows posttetanic depression. produce a movement.
GLOSSARY G-2.5
......................................
protease Enzymes that break down peptide bonds of proteins and quantal release The release of neurotransmitter in discrete packets
polypeptides. which correspond to vesicles containing transmitter
protein kinase Any enzyme that catalyzes the transfer of a molecules.
phosphate group from ATP to a protein, creating a phospho- quantal synaptic transmission The concept that neurotrans-
protein. mitter is released in multiples of discrete "packets." It is now
proteins Large molecules composed of one or more chains of al- apparent that the quantal packets represent individual presy-
pha amino acid residues (i.e., polypeptide chains). naptic vesicles.
proteolysis The splitting of proteins by hydrolysis of peptide quaternary structure The characteristicways in which the subunits
bonds. of a protein containing more than one polypeptide chain are
proteolytic Protein-hydrolyzing. combined.
prothoracic glands Ecdysone-secreting tissues situated in the an- r selection A pattern of energetic investment in reproduction in
terior thorax of insects. which large numbers of very small offspring are produced,
prothoracicotropic hormone (MTH)A neurohormone produced each with a low chance of survival due to lack of parental
by neurosecretory cells in the pars intercerebralis of the brain. care.
PTTH activates the prothoracic gland to synthesize and se- radial finks Extensions from peripheral doublets to the central
crete molting hormones. sheath in cilia and flagella.
proximal tubules Coiled portions of the renal tubules located radiation, thermal The transfer of heat by electromagnetic radia-
in the renal cortex, beginning at the glomerulus and leading tion without direct contact between objects.
to (and continuous with) the descending limb of the loop of radioimmunoassays (RIAs)An immunological technique for the
Henle. measurement of minute quantities of antigen or antibody, hor-
pseudopodium Literally, false foot; a temporary projection of an mones, certain drugs, and other substances with the use of ra-
amoeboid cell for engulfment of food or for locomotion. dioactively labeled reagents.
pseudopregnancy A false pregnancy. radioisotope A radioactive isotope.
psychophysics The branch of psychology concerned with rela- radula A rasp-like structure in the mouth of many gastropods.
tionships between physical stimuli and perception. range fractionation The pattern in which receptors within one sen-
pulmonary Pertaining to or affecting the lungs. sory modality are tuned to receive information within rela-
pupa A developmental stage of some insect groups; between the tively narrow, but not identical, intensity ranges, so the entire
larva and the adult. dynamic range of the modality is divided among different
pupil The opening at the center of the iris through which light classes of receptors. For example, in the human eye rods re-
passes into the eye. spond to dim light but are saturated in bright light; cones are
purinergic Referring to nerve endings that release purines or their less sensitive to dim light but remain responsive in bright light.
\ derivatives as transmitter substances. rate constant (specificreaction rate) The proportionality factor by
purines A class of nitrogenous heterocyclic compounds, C5H4N4, which the concentration of a reactant in an enzymatic reac-
derivatives of which (purine bases) are found in nucleotides; tion is related to the reaction rate.
they are colorless and crystalline. reactive hyperernia Higher than normal blood flow that occurs fol-
pyloric Pertaining to the caudal portion of the vertebrate stomach lowing a brief period of ischemia.
where it joins the small intestine. receptive field That area of an organism's body (e.g., on the skin
pyloric sphincter Sphincter guarding the opening of the stomach or in the retina) that when stimulated influences the activity of
into the small intestine. a given neuron is the receptive field of that neuron.
pylorus The distal stomach opening, ringed by a sphincter, that re- receptor Molecules that are situated on a membrane and that in-
leases the stomach contents into the duodenum. teract specifically with messenger molecules, such as hor-
pyramidal tract A bundle of nerve fibers originating in the motor mones or transmitters.
cortex and descending down the brain stem to the medulla receptor cell A neuronal cell that is specialized to respond to some
oblongata and to the spinal cord; responsible for mediating particular sensory stimulation.
control of voluntary muscle movements. receptor current A stimulus-induced change in the movement of
pyrimidine A class of nitrogenous heterocyclic compounds, ions across a receptor cell membrane.
C4H4N,, derivatives of which (pyrimidine bases) are found in receptor-mediated endocytosis Specialized process of endocytosis
nucleotides. that requires solutes being transported to temporarily bind to
pyrogen Substance that leads to a resetting of a homeotherm's receptor molecules embedded in the cell membrane prior to
body thermostat to a higher set point, thereby producing transport across the cell membrane.
fever. receptor molecules Molecules that are situated on the outer sur-
face of the cell membrane and that interact specifically with
Q,, The ratio of the rate of a reaction at a given temperature to its messenger molecules, such as hormones or neurotransmitters.
rate at a temperature 10°C lower. receptor potential A change in membrane potential elicited in sen-
QRS-wave That portion of the electrocardiogram related to de- sory receptor cells by sensory stimulation, which changes the
polarization of the ventricle. flow of ionic current across the cell membrane.
quality A property that distinguishes sensory stimuli within a sen- receptor tyrosine kinases (RTKs) Receptors with intrinsic
sory modality; e.g., color is a quality of visual stimuli. tyrosine kinase activity, which are known to bind insulin and
quantal content The number of neurotransmitter molecules in one a number of growth factors. When activated by external
synaptic vesicle. signal binding, RTKs transfer the phosphate group from
G-26 GLOSSARY
.........................................
ATP to the hydroxyl group on a tyrosine residue of selected filtrate per unit time. Total renal clearance is the amount of
proteins in the cytosol. RTKs also phosphorylate themselves ultrafiltrate produced by the kidney per unit time.
when activated; this autophosphorylation enhances the activ- renin A proteolytic enzyme produced by specialized cells in renal
ity of the kinase. arterioles; converts angiotensinogen to angiotensin.
reciprocal inhibition Inhibition of the motor neurons innervating rennin An endopeptidase enzyme that coagulates milk by pro-
one set of muscles during the reflex excitation of their moting the formation of calcium caseinate from the milk pro-
antagonists. tein casein; found especially in the gastric juice of young
recombinant DNA An engineered DNA molecule resulting from mammals.
the two different organisms. Renshaw cells Small inhibitory interneurons in the ventral horn
recruitment The pattern in which neurons with increasingly that are excited by branches of a-motor neuron axons that
high thresholds become active as intensity increases. This pat- feed back on the motor neuron pool.
tern can occur in sensory neurons (range fractionation) or in repolarization The return to resting polarity of a cell membrane
motor neurons. that has been depolarized.
rectal gland Organ near the rectum of elasmobranchs that excretes repression proteins Proteins that can bind to a short region of
a highly concentrated NaCl solution. DNA preceding the structural geneis), thus preventing tran-
redox pair Two compounds, molecules, or atoms involved in mu- scription.
tual reduction and oxidation. repressor gene (regulator gene) A gene that produces a substance
reductant Donor of electrons in a redox reaction. (repressor) that shuts off the structural-gene activity of an
reduction The addition of electrons to a substance. operon by an interaction with its operator gene.
reduction potential A measurement of the tendency of a reducer reserpine A botanically derived tranquilizing agent that interferes
to yield electrons in a redox reaction, expressed in volts. with the uptake of catecholamine from the cytosol by secre-
reflex An action that is generated without the participation of the tory vesicles; its effect is to deplete the catecholamine content
highest neuronal centers and is thus not voluntary; an invol- of adrenergic cells.
untary motor response mediated by a neuronal arc in re- residual volume The volume of air left in the lungs after maximal
sponse to sensory input. expiratory effort.
reflex arc A neuronal pathway that connects sensory input and resistance (R)The property that hinders the flow of current. The
motor output; consists of afferent nerve input to a nerve cen- unit is the ohm (a), defined as the resistance that allows 1am-
ter that produces activity in efferent nerves to an effector pere (A)of current to flow when a potential drop of 1volt (V)
organ. exists across the resistance. It is equivalent to the resistance of
refraction The bending of light rays as they pass from a medium a column of mercury 1 mm2in cross-sectional area and 106.3
of one density into a medium of another density. cm long. R = p x length + cross-sectional area.
refractive index The refractive power of a medium compared with resistivity (p) The resistance of a conductor 1 cm in length and
that of air, designated 1. 1 cm2 in cross-sectional area.
refractory period The period of increased membrane threshold im- respiratory acidosis A decrease in blood pH associated with a fall
mediately following an action potential. Absolute refractory in blood PCO, as a result of lung hypoventilation.
period: The initial phase of the refractory period when no AP respiratory alkalosis An increase in blood pH associated with a
can be generated. Relative refractory period: The later phase rise in blood PCOz as a result of lung hyperventilation.
of the refractory period when the threshold is elevated, but an respiratory chain See electron-transport chain.
AP can be generated with sufficiently intense stimulation. respiratory pigment A substance that combines reversably with
regenerative Self-reinforcing; utilizing positive feedback; auto- oxygen-for example, hemoglobin.
catalytic. respiratory quotient (RQ)The ratio of CO, production to 0, con-
regulator Animals that use biochemical, physiological, behavioral, sumption; depends on type of food oxidized by the animal.
and other mechanisms to maintain internal homeostasis. respirometry Measurement of an animal's respiratory exchange.
regulator gene Genes that code for repressor proteins, which sup- resting potential The normal, unstimulated membrane potential
press the action of structural genes. of a cell at rest.
regurgitation Reverse movement of intestinal luminal contents rete mirabile An extensive countercurrent arrangement of arter-
produced by reverse peristalsis. ial and venous capillaries.
Reissner's membrane A membrane within the mammalian reticulum A small network.
cochlea. retina The photosensitive inner surface of the vertebrate eye.
release-inhibitinghormone (RM;release-inhibitingfactor, RIF) A retinal The aldehyde of retinol obtained from the enzymatic ox-
hypothalamic neurosecretion carried by portal vessels to the idative cleavage of carotene; in the 11-cis form it unites with
adenohypophysis, where it restrains the release of a specific opsins in the retina to form the visual pigments.
hormone. retinal streak (visualstreak) A retinal structure-found in the eyes
releasing hormone (RH, releasing factor) A hypothalamic neu- of some species that inhabit plains-in which photoreceptors
rosecretion that stimulates the liberation of a specific hormone are packed into a horizontal streak across the retina, provid-
from the adenohypophysis. ing high resolution along the visual horizon. This pattern is
releasing stimulus (key stimulus) The stimulus that is effective in similar to the fovea of primates, but it has a different shape.
producing a fixed action pattern. retinol Vitamin A (C,,H,,O), an alcohol of 20 carbons; converted
renal clearance That volume of plasma containing the quantity of reversibly to retinal by enzymatic dehydrogenation.
a freely filtered substance that appears in the glomerular retinular cell A photoreceptor cell of the arthropod compound eye.
GLOSSARY G-27
......................................
reversal potential The membrane potential at which no current salivary glands Glands that secrete saliva into the headgut.
flows through membrane ion channels, even though the chan- saltatory (conduction)Jumping; discontinuous.
nels are open; it is equal to the equilibrium potential for the salt glands Osmoregulatory organs of many birds and reptiles that
ion or ions that are conducted through the open channels. live in desert or marine environments. A hypertonic aqueous
Reynolds number (Re) A unitless number; the tendency of a flow- exudate is formed by active salt secretion into the small
ing gas or liquid to become turbulent is proportional to its ve- tubules situated above the eyes and is excreted via the nostrils.
locity and density and inversely proportional to its viscosity. "salting out" A decrease in Bunsen solubility coefficient as a result
Calculated from these parameters, the Reynolds number in- of increased ionic strength of the solvent.
dicates whether flow will be turbulent or laminar under a par- sarcolemma The surface membrane of muscle fibers.
ticular set of conditions. sarcomere The contractile unit of a myofibril, it is bounded by two
rhabdome The aggregate structure consisting of a longitudinal Z disks.
rosette of rhabdomeres located axially in the ommatidium. sarcoplasm Cytosol of a muscle cell.
rhabdomere The light-absorbing part of a retinular cell that faces sarcoplasmic reticulum (SR) A smooth, membrane-limited net-
the central axis of an ommatidium; the photopigment-bear- work surrounding each myofibril. Calcium is stored in the SR
ing surface membrane is expanded into closely packed mi- and released as free Ca2+during muscle excitation-contrac-
crovilli, increasing the amount of photosensitive membrane. tion coupling.
rheogenic Producing electric current. sarcotubular system The sarcoplasmic reticulum plus the trans-
rhodopsin (visual purple) A purplish red, light-sensitive chromo- verse tubules.
protein with 11-cisretinal as its prosthetic group; found in the saturated In reference to fatty acid molecules, indicates that the
rods and cones of the retina; bleaches to "visual yellow" (all- carbon-carbon bonds are single, with each carbon atom bear-
trans-retinal) when it absorbs incident light. ing two hydrogens. Without free valence electrons.
ribonucleic acid See RNA. scala media The cochlear duct, a membranous labyrinth contain-
ribose A pentose monosaccharide with the chemical formula ing the organ of Corti and the tectorial membrane; it is filled
HOCH,(CHOH),CHO; a constituent of RNA. with endolymph.
ribosome Ribonucleoprotein particles found within the cytoplasm; scala tympani A cochlear chamber connected with the scala
the sites of intersection of mRNA, tRNA, and the amino acids vestibuli through the helicotrema; it is filled with perilymph.
during the synthesis of polypeptide chains. scala vestibuli A cochlear chamber beginning in the vestibule, con-
rigor mortis The rigidity that develops in dying muscle as ATP be- necting with the scala tympani through the helicotrema; it is
comes depleted and cross bridges remain attached. filled with perilymph.
Ringer solution Physiological saline solution. scaling The study of how both anatomical and physiological char-
RNA (ribonucleic acid) A nucleic acid made up of adenine, gua- acteristics change with body mass.
nine, cytosine, uracil, ribose, and phosphoric acid; responsi- Schwann cell A neuroglial cell outside the central nervous system
ble for the transcription of DNA and the translation into that wraps its membrane around axons during development
protein. to produce the insulating myelin sheath that envelops periph-
rods One class of vertebrate visual receptor cells, the cones being eral axons in the regions between nodes of Ranvier.
the other; very sensitive to light, based on cellular physiology scintillation counter An instrument that detects and counts tiny
and on a high degree of convergence onto second-order cells. flashes of light produced in scintillation fluid produced by par-
In most species, there is only one class of rods in the retina, so ticles emitted from radioisotopes.
rods cannot convey information about color. SDA (specificdynamic action) The increment in metabolic energy
Root effect (Root shift) A change in blood oxygen capacity as a re- cost that can be ascribed to the digestion and assimilation of
sult of a pH change. food; it is highest for proteins.
round window A membrane-covered opening, separating the rnid- secondary structure Refers to the straight or helical configuration
dle ear and the cochlea, through which pressure waves leave of polypeptide chains.
after traveling through the cochlea. secondary vacuole Vacuole formed when food containing vacuole
rumen The storage and fermentation chamber in the digastric merges with enzyme-containing lysosomes.
stomach of ruminants. second law of thermodynamics All natural or spontaneous
rumination The chewing of partially digested food brought up by processes are accompanied by an increase in entropy.
reverse peristalsis from the rumen in ungulate animals and in second messenger A term applied to CAMP,cGMP, Ca2+,or any
other ruminants. other intracellular regulatory agent that is itself under the con-
saccharide A family of carbohydrates that includes the sugars; they trol of an extracellular first messenger, such as a hormone.
are grouped as to the number of saccharide (C,H,,O,-I) second-order enzyme kinetics Describes enzymatic reactions
groups comprising them: the mono-, di-, tri-, and polysaccha- whose rates are determined by the concentrations of two re-
rides. actants multiplied together or of one reactant squared.
sacculus One of the vertebrate organs of equilibrium. second-order neuron A neuron that receives input from primary
safety factor A factor relating input and output in a system, de- sensory neurons.
scribing how likely transmission through the system will fail; secretagogue A substance that stimulates or promotes secretion.
the higher the safety factor, the less likely it is that transmis- secretin A polypeptide hormone secreted by the duodenal and je-
sion will fail. junal mucosa In response to the presence of acid chyme in the
saliva A water-like fluid secreted in the upper alimentary canal intestine; induces pancreatic secretion into the intestine and is
(headgut);aids in mechanical and chemical digestion. chemically identical to enterogastrone.
G-28 GLOSSARY
.........................................
secretory granules (secretory vesicles) Membrane-bound cyto- signal-to-noise ratio The relation between a signal and the
plasmic granules containing secretory products of a cell. random background activity that arises as the result of kinetic
segmentation Rhythmic contractions of the circular muscle layer energy or other irrelevent events; information theory has thor-
of the intestine that mixes the intestinal contents. oughly considered this relation and generated rules that de-
selectivity sequence See affinity sequence. scribe efficient and effective information transfer.
self-repair capability Ability of cell organelles and membranes single-unit muscle A smooth muscle in which individual fibers are
to reseal themselves when chemically or mechanically dis- coupled through gap junctions, allowing excitation to spread
turbed. through the muscle independent of neuronal activity; con-
semicircular canals Three of the vertebrate organs of equilibrium, traction in these muscles is often myogenic, driven by internal
which sense acceleration of the body with respect to the grav- pacemaker cells.
itational field. sinoamal node A mass of specialized cardiac tissue that lies at the
seminal vesicles Paired sacs attached to the posterior urinary blad- junction of the superior vena cava with the right atrium;
der that have tubes joining the vas deferens in the male. it acts as the pacemaker of the heart in initiating each cardiac
semipermeable membrane A membrane that allows certain mol- contraction.
ecules but not others to pass through it. sinus A cavity or sac; a dilated part of a blood vessel.
sensation The perception of a sensory stimulus (as opposed to sinus node (sinoatrial node, SA node) The junction between the
the reception of stimulus energy in a primary sensory re- right atrium and the vena cava, the location of the pacemaker.
ceptor). sinus venosus The membrane chamber attached to the heart that
sensilla (plural; sensillum, singular) Collections of sensory recep- receives Venus blood in fish, amphibians, and reptiles, and
tors in the periphery of an organism, usually an invertebrate; transmits it to the atrium.
sensilla are typically very simple in structure, lacking acces- skeletal muscle The striated muscle whose contraction is respon-
sory structures. sible for moving the bodies of animals. Contraction of these
sensillum A chitinous, hollow, hairlike projection of the arthropod fibers is neurogenic; that is, contraction occurs only when the
exoskeleton that serves as an auxiliary structure for sensory fibers are excited by synaptic input from motor neurons.
neurons. sliding-filament theory The theory that muscle sarcomeres shorten
sensor Mechanical, electrical, or biological device that detects when actin thin filaments are actively pulled toward the mid-
changes in its immediate environment. dle of myosin thick filaments by the action of myosin cross-
sensory adaptation The property of sensory systems to become bridges.
less sensitive to stimuli during prolonged or repeated sliding-tubule hypothesis Bending movements of cilia and flagella
stimulation. are produced by active longitudinal sliding of the axonemal
sensory fiber An axon that carries sensory information to the cen- microtubules past one another.
tral nervous system. slow chemical synaptic transmission Synaptic transmission at a
sensory filter network Neuronal circuits that selectively transmit chemical synapse mediated by neurotransmitters that bind on
some features of a sensory input and ignore other features. the postsynaptic membrane to receptor molecules that affect
sensory modality A set of sensory structures that are tuned to re- intracellular second messenger systems, typically through G
ceive a particular class of energy; e.g., vision, hearing, and the proteins. Binding of the transmitter to the receptor complex
sense of smell are three sensory modalities. activates the second messengers, which in turn modify the
sensory reception The absorption of stimulus energy by a neuron state of ion channel proteins.
that produces a receptor potential in the neuron and that may smooth muscle Muscle without sarcomeres and hence without
produce action potentials that travel to the central nervous striations. Myofilaments are nonuniformly distributed within
system. small, mononucleated, spindle-shaped cells.
series elastic components (SEC)Elastic structures ( such as tendons sodium activation An increased conductance of excitable mem-
and other connective tissues) that are arranged in series with branes to sodium ions in response to membrane depolariza-
the contractile elements in muscle. tion; believed to result from an opening of sodium gates
serosa Outermost layer of alimentary tract. associated with membrane channels.
serosal Pertaining to the side of an epithelial tissue facing the sodium hypothesis The upstroke of an action potential is due to
blood, as opposed to the mucosal side, which faces the exte- an inward movement of Na+ down its electrochemical gradi-
rior or luminal space. ent as a result of a transient increase in sodium permeability.
serotonin 5-Hydroxytryptamine, 5-HT; a neurotransmitter, sodium inactivation Loss of responsiveness of sodium gates to de-
CIOH12N2O. polarization; develops with time during a depolarization and
serum The clear component of blood plasma. persists for a short period after repolarization of the
servo mechanism A control system that utilizes negative membrane.
feedback to correct deviations from a selected level, the set sodium-potassium ATPase (sodium pump) Membrane protein re-
point. sponsible for maintaining the asymmetrical concentrations of
set point In a negative-feedback system, the state to which feed- Na+ and K+ions across the cell membrane; it actively ex-
back tends to bring the system. trudes Na+ from the cell and takes up K+ from extracellular
siemen (S) The unit of electrical conductance; reciprocal of the fluids at the expense of metabolic energy. In some sodium
ohm. pumps, there is a 3 :2 exchange of intracellular Na+ for ex-
sign stimulus The most basic essential pattern of sensory input re- tracellular K+.
quired to release an instinctual pattern of behavior. sol The low-viscosity state of cytoplasm.
GLOSSARY G-29
......................................
solvation The process of dissolving a solute in a solvent; hydration, of salt across the portions of the epithelial cell membranes fac-
or clustering, of water molecules around individual ions and ing intercellular clefts.
polar molecules. standing wave A resonating wave with fixed nodes.
solvent drag Process in which smaller solute molecules are carried stapes (stirrup) The innermost auditory ossicle, which articulates
passively along with the water as it flows down its osmotic at its apex with the incus and whose base is connected to the
gradient through hydrated channels. oval window.
soma The nerve cell body, or perikaryon; in general, the body. starch A polysaccharide of plant origin, formula (C,H,,O,), .
somatic Referring to the body tissues as distinct from the germ Starling curves Curves that describe the relationship between heart
cells. work and filling pressure.
somatic system The part of the nervous system that receives input statocyst Gravity-sensing sensory organ made up of mechanore-
from the body. ceptive hair cells and associated particles called statoliths.
somatosensory cortex The region of the cerebral cortex that re- statolith A small, dense, solid granule found in statocysts.
ceives sensory input from the body surface. steady state Dynamic equilibrium.
somatostatin Growth-hormone-inhibiting hormone, which in- stenohaline Able to tolerate only a narrow range of salinities.
hibits growth-hormone release from the pituitary. stereocilia Nonmotile filament-filled projections of hair cells; these
spatial summation Integration by a postsynaptic neuron of simul- "hairs" lack the internal structure of motile "9 + 2" cilia.
taneous synaptic currents that arise from the terminals of dif- steric Pertaining to the spatial arrangement of atoms.
ferent presynaptic neurons.
steroid hormones Cyclic hydrocarbon derivatives synthesized
specific dynamic action (SDA)A marked increase in metabolism
from cholesterol.
accompanying the digestion and assimilation of food.
sterols A group of solid, primarily unsaturated polycyclic alcohols.
specific resistance (R,) Resistance per unit area of a membrane in
stimulus A substance, action, or other influence that when applied
ohms per square centimeter.
with sufficient intensity to a tissue causes a response.
spectrophotometer A device that passes a beam of visible or W
stomach The major digestive region of the alimentary canal.
light through a fluid-filled vial and measures the emerging
stomatogastric ganglion A ganglion in Crustacea that contains the
wavelengths.
central pattern generator neurons that control the rhythmic
spectrum Specific, charted bands of electromagnetic radiation
activity of the stomach and associated organs.
wavelengths produced by refraction or diffraction.
STP See standard temperature and pressure.
sphincter A ring-shaped band of muscle fibers capable of con-
stricting an opening or a passageway. stretch receptor Sensory receptors that respond to stretch, typically
associated with lungs or muscle tissue.
sphingolipid A lipid formed by a fatty acid attached to the nitro-
gen atom of sphingosine, a long-chain, oily amino alcohol stria vascularis Vascular tissue layer over the external wall of the
(C18H,,0,N). The sphingolipids occur primarily in the mem- scala media; secretes the endolymph.
branes of brain and nerve cells. striated muscle Characterized by sarcomeres aligned in register.
spike-initiating zone Region of the nerve axon where an action po- Skeletal and cardiac muscle are striated.
tential is initiated. In many-but not all-neurons, the axon stroke volume The volume of blood pumped by one ventricle dur-
hillock. ing a single heartbeat.
spinal canal The fluid-filled cavity that runs longitudinallythrough structural gene A gene coding for the sequence of amino acids that
the vertebrate spinal cord; it is confluent with the cerebral make up a polypeptide chain.
ventricles. strychnine A poisonous alkaloid (C,,H,,N,O,) that blocks in-
spinal cord The portion of the vertebrate central nervous system hibitory synaptic transmission in the vertebrate central ner-
that is encased in the vertebral column, extending from the vous system.
caudal end of the medulla oblongata to the upper lumbar re- submucosa Second-most inner layer of alimentary canal, underly-
gion; constructed of a core of gray matter and an outer layer ing inner mucosa.
of white matter. submucosal plexus Neural plexus that acts to stimulate gut motil-
spinal root A large bundle of axons that enters or leaves the spinal ity and secretion.
cord at each spinal segment. substrate A substance that is acted on by an enzyme.
spindle organ A stretch receptor of vertebrate skeletal muscle. substrate-level phosphorylation Mechanism by which chemical
spiracle Surface opening of the tracheal system in insects. energy of oxidation is stored in the form of ATP.
spiral ganglion The ganglion lying near the cochlea that contains succus entericus Digestive juice secreted by the glands of
the somata of neurons whose axons carry auditory informa- Lieberkiihn in the small intestine.
tion from the hair cells of the organ of Corti to the auditory sulfhydryl group The radical -SH.
centers of the brain. supercooling Cooling of a fluid below its freezing temperature
spirometry Measurement of an animal's 0, and CO, turnover. without actual freezing because of the failure of ice crystals
standard metabolic rate (SMR)Similar to basal metabolic rate, but fail formation.
utilized for metabolic rate of a heterotherm maintained at a supraoptic nucleus A distinct group of neurons in the hypothala-
selected body temperature. mus, just above the optic chiasma; their neurosecretory end-
standard temperature and pressure (STP, dry, STPD) 2S°C, 1at- ings terminate in the neurohypophysis.
mosphere (atm). surface charge Electric charge at the membrane surface, aris-
standing-gradient hypothesis Hypothesis describing process of ing from fixed charged groups associated with the membrane
solute-coupled water transport, involving the active transport surface.
G-30 GLOSSARY
...................................... ......................................
surface hypothesis Hypothesis of Rubner that metabolic rate of tachycardia An increase in heart rate above the normal level.
birds and mammals should be proportional to body surface target cells Cells that preferentially bind and respond to specific
area. hormones.
surface tension The elasticity of the surface of a substance (par- taxis Locomotion that is oriented with respect to a stimulus di-
ticularly a fluid), which tends to reduce the surface area at rection or gradient.
each interface. tectorial membrane A fine gelatinous sheet lying on the organ of
surfactant A surface-active substance that tends to reduce surface Corti in contact with the cilia of cochlear hair cells.
tension, for example, in the lung. tectum The highest center for processing visual information in
swim bladder A gas-filled bladder used for flotation; found in fishes and amphibians.
many teleost fishes. teleost Bony fish, of the infraclass Teleostei.
sympathetic nervous system Thoracolumbar part of the auto- temporal Referring to the lateral areas of the head above the zy-
nomic nervous system; increased activity in sympathetic neu- gomatic arch. Also, relating to time; time limited.
rons typ~cally provides metabolic support for vigorous temporal lobe A lobe of the cerebral hemisphere, situated in the
physical activity, so this system has been called "the fight or lower lateral area, at the temples.
flight system." temporal summation Summation of postsynaptic membrane
symports Coupled membrane transporters that transfer two potentials that occur close to one another in time, but not si-
solutes in the same direction across the cell membrane. multaneously.
synapse A specialized area that connects two directly interacting tendon A band of tough fibrous connective tissue that anchors a
nerve cells. At a synapse, activity in the presynaptic (trans- skeletal muscle to the skeleton, allowing contraction of the
mitting) cell influences the activity of the postsynaptic (re- muscle to move the body of an animal.
ceiving)cell.
terminal cisternae The closed spaces that make up part of the sar-
synaptic cleft The space separating the cells at a synapse. coplasmic reticulum on both sides of the Z line, making close
synaptic current The ionic current that flows across a postsynap- contact with the T tubules.
tic membrane when ion channels open after neurotransmitter tertiary structure Refers to the way a polypeptide chain is folded
molecules bind to membrane receptors; the flow of ions causes or bent to produce the overall conformation of the molecule.
the membrane potential to change toward the equilibrium po- testosterone A steroid androgen synthesized by the testicular in-
tential of the ion or ions that flow through the synaptic
terstitial cells of the male; responsible for the production and
channels.
maintenance of male secondary sex characteristics.
synaptic delay The time separating the arrival of an impulse at a
tetanus An uninterrupted muscular contraction caused by high fre-
presynaptic nerve terminal and a change in the membrane po-
quency motor impulses. Also the name of a neurotoxin that is
tential of the postsynaptic cell.
retrogradely (toward the cell body) transported in axons and
synaptic efficacy Effectiveness of a presynaptic impulse in pro- that causes prolonged excitation of muscle fibers, causing
ducing a postsynaptic potential change. tetanic contraction.
synaptic facilitation Increase in synaptic efficacy. tetraethylammonium (TEA) A quarternary ammonium agent,
synaptic inhibition A change in a postsynaptic cell that reduces the (C,H,),N, that can be used to block some potassium channels
probability of its generating an action potential; produced by in membrane.
a transmitter substance that elicits a postsynaptic current hav- tetrodotoxin (TTX) The pufferfish poison, which selectively
ing a reversal potential more negative than the threshold for blocks voltage-gated sodium ion channels in the membranes
the action potential. of excitable cells.
synaptic noise Irregular changes in the transmembrane potential thalamus A major center in the midbrain of birds and mammals
of a postsynaptic cell, produced by random subthreshold that receives and transmits both sensory and motor informa-
synaptic input. tion.
synaptic transmission Transfer of a signal between two theca interna The internal vascular layer encasing an ovarian
cells, usually between a neuron and another cell, which follicle; responsible for the biosynthesis and secretion of
could be a neuron, a muscle, or some other effector cell such estrogen.
as a gland.
theophylline A crystalline alkaloid (C,H,N,H,O) found in tea; in-
synaptic vesicles Membrane-bound vesicles containing neuro- hibits the enzyme phosphodiesterase, thereby increasing the
transmitter molecules; located within axon terminals. level of CAMP;also releases Ca2+from calcium-sequestering
syncitium A network of cells that are connected by low- organelles.
resistance intercellular pathways. thermal conductance Ability of a material to conduct heat, which
syneresis Contraction of a gelled mixture so that a liquid is is poor in insulating materials such as fur or feathers.
squeezed out from molecular interstices. thermal neutral zone (thermoneutral zone) That range of ambient
systemicPertaining to or affecting the body; for example, systemic temperatures within which a homeotherm can control its tem-
circulation. perature by passive measures and without elevating its meta-
systole The portion of the cardiac sequence when the heart mus- bolic rate to maintain thermal homeostasis.
cle is contracting; it takes place between the first and second thermogenesis The production of body heat by metabolic means
heart sounds as the blood flows through the aorta and pul- such as brown-fat metabolism or muscle contraction during
monary artery. shivering.
T-wave That portion of the electrocardiogram associated with re- thermophilic behavior Heat-seeking behavior.
polarization (and usually relaxation) of the ventricle. thermophobic behavior Heat-avoiding behavior.
thermoreceptor Sensory nerve ending specifically responsive to transcellular pathways Routes through or around a cell by which
temperature changes. substances are actively transported across an epithelium.
thick filament A myofilament made of myosin. transcription The formation of an RNA chain of a complementary
thin filament A myofilament that contains actin and regulatory base sequence from the informational base sequence of DNA.
proteins. transducer A mechanism that translates energy or signals of one
thoracic cage The chest compartment formed by the ribs and di- form into a different kind of energy or signals.
aphragm containing the lungs and heart. transducer molecule Intermediate molecule (G protein) within the
thoracic duct Duct draining the lymphatic system into the anterior cell membrane that transmits a hormone-initiated signal from
cardural vein. the externally facing hormone receptor to the internally fac-
3-dehydroretinal A derivative of vitamin A that is found in the ing enzyme.
visual pigments of freshwater fishes and amphibia. transducin The G protein that links the capture of light by
3,5,3-triiodothyronine An iodine-bearing tyrosine derivative syn- rhodopsin molecules with a change in the current flowing
thesized in and secreted by the thyroid gland; raises cellular across the membrane of photoreceptors.
metabolic rate, as does thyroxine. transduction General term for the modulation of one kind of en-
threshold potential The potential at which a response (e.g., an ac- ergy by another kind of energy. Thus, sense organs transduce
tion potential or a muscle twitch) is produced. sensory stimuli into nerve impulses.
threshold stimulus The minimum strength of stimulation nec- transfer RNA (TRNA)A small RNA molecule that is responsible
essary to produce a detectable response or an all-or-none for the transfer of amino acids from their activating enzymes
response. to the ribosomes; there are 20 tRNAs, one for each amino
thymine A pyrimidine base, 5-methyluracil (C,H6N,0), a con- acid.
stituent of DNA. transgenic animal An animal in which its genetic constitution has
thyroid-stimulating hormone (TSH)An adenohypophyseal hor- been experimentally altered by the addition or substitution of
mone that stimulates the secretory activity of the thyroid genes from other animals of that species or other species.
gland. translation Utilization of the DNA base sequence for linear orga-
thyroxine An iodine-bearing, tyrosine-derived hormone that is nization of amino acid residues on a polypeptide; carried out
synthesized and secreted by the thyroid gland; raises cellular by mRNA.
metabolic rate. transmitter substance A chemical mediator liberated from a presy-
tidal volume The volume of air moved in or out of the lungs with naptic ending, producing a conductance change or other re-
each breath. sponse in the membrane of the postsynaptic cell.
tight junction An area of membrane fusion between adjoining transmural pressure The difference in pressure across the wall of
cells; prevents passage of extracellular material between ad- a structure, for example, a blood vessel.
joining cells; prevents passage of extracellular material be- transphosphorylation The transfer of phosphate groups between
tween the cells. organic molecules, bypassing the inorganic phosphate stage.
time constant (t) A measure of the rate of accumulation or decay transverse tubules ( T tubules) Branching membrane-limited, in-
in an exponential process; the time required for an exponen- tercommunicating tubules that are continuous with the sur-
tial process to reach 63% completion. In electricity, it is pro- face membrane and are closely apposed to the terminal
portional to the product of resistance and capacitance. cisternae of the sarcoplasmic reticulum.
tonic Steady; slowly adapting. traveling wave A wave that moves through the propagating
tonicity (hyper-, hypo-, iso-) The relative osmotic pressure of a so- medium, as opposed to a standing wave, which remains
lution under given conditions (e.g., its osmotic effect on a cell stationary.
relative to the osmotic effect of plasma on the cell). tricarboxylic acid cycle (TCA cycle; Krebs cycle; citric acid cycle)
tonotopic map A pattern of auditory projection to the brain in The metabolic cycle responsible for the complete oxidation of
which neurons are arranged and make synapses based on the the acetyl portion of the acetyl coenzyme A molecule.
frequency of the sound to which they respond. trichromacy theory The theory that three kinds of photore-
tonus Sustained resting contraction of muscle, produced by basal ceptor cone cells exist in the human retina, each with a char-
neuromotor activity. acteristic maximal sensitivity to a different part of the color
c torpor A state of inactivity, often with lowered body temperature spectrum.
and reduced metabolism, that some homeotherms enter into triglyceride A neutral molecule composed of three fatty acid
SO as to conserve energy stores. residues esterified to glycerol; formed in animals from carbo-
trachea The large respiratory passageway that connects the phar- hydrates.
ynx and bronchioles in the vertebrate lung. triglycerol A neutral molecule composed of three fatty acid
tracheal system Consists of air-filled tubules that carry respiratory residues esterified to glycerol; formed in animals from carbo-
gases between the tissues and the exterior in insects. hydrates.
tracheoles Minute subdivisions of the tracheal system of insects. trirner A compound made up of three simpler identical molecules.
tragus The tab that extends from the ventral (anterior) edge of the trimethylamine oxide A nitrogenous waste product, probably
outer ear and partially covers the opening of the ear. from choline decomposition.
train of impulses A rapid succession of action potentials propa- tritium A radioactive isotope of hydrogen with an atomic mass of
gated down a nerve fiber. three (H3).
trans A configuration with particular atoms or groups on opposite Triton X-100 A nonionic detergent used in cell biology to solubi-
sides. lize lipids and certain cell proteins.
G-32 GLOSSARY
......................................
trituration Grinding; mastication. some of the carbon-carbon bonds are double. Having free va-
tRNA See transfer RNA. lence electrons.
trophic level Individual level within a food chain. upper critical temperature (UCT)Ambient temperature beyond
trophic substances Chemical substances believed to be released which normal heat loss mechanisms cannot prevent increase
from neuron terminals and to influence the chemical and func- in body temperature.
tional properties of the postsynaptic cell. uracil A pyrimidine (C,H,O,N,) constituent of RNA.
tropomyosin A long protein molecule located in the grooves of urea (NH,),CO, the primary nitrogenous waste product in the
actin filaments of muscle; inhibits muscle contraction by urine of mammals.
blocking the interaction of myosin cross bridges with actin ureotelic Pertaining to the excretion of nitrogen in the form of
filaments. urea.
troponin A complex of globular calcium-binding proteins associ- ureter A muscular tube passing urine to the bladder from the
ated with actin and tropomyosin in the thin filaments of kidney.
muscle. When troponin binds Ca2+, it undergoes a con- urethra The channel passing urine from the bladder out of the
formational change, allowing tropomyosin to reveal myosin- body.
binding sites on the actin filament. uric acid A crystalline waste product of nitrogen metabolism
trypsin An enzyme specifically attacking peptide bonds in which found in the feces and urine of birds and reptiles; poorly sol-
the carboxyl group is provided by arginine or lysine. uble in water.
trypsinogen A proenzyme of trypsin. uricolytic See uricotelic.
tubulin An actinlike, 4 nm globular protein molecule that is the uricolyticpathway The pathway through which uric acid or urates
building block of microtubules. are cleavaged.
tunica adventitia The fibrous outer layer of arterial blood vessel uricotelic Pertaining to the excretion of nitrogen in the form of uric
walls. acid.
tunica intima The inner lining of arterial blood vessel walls. Ussing chamber A chamber used to suspend tissue such as frog
tunica media The middle layer of arterial blood vessel walls con- skin and measure its epithelial transport properties.
sisting of smooth muscle and elastic tissue. utriculus One of the vertebrate organs of equilibrium.
turbinates Chambers of the nasal passages with olfactory recep-
tors in the surface epithelium. vacuole A membrane-limited cavity in the cytoplasm of a cell.
turbulent flow Flow pattern in which exists sharp gradients and vagus nerve (tenth cranial nerve) A major cranial nerve that sends
inconsistenciesin velocity and direction of flow fluid. sensory fibers to the tongue, pharynx, larynx, and car; motor
fibers to the esophagus, larynx, and pharynx; and parasym-
turgor Distension; swollenness.
pathetic and afferent fibers to the viscera of the thoracic and
twitch muscle (fast muscle) The most common striated vertebrate
abdominal regions.
skeletal muscle type, usually pale in color because of its low
myoglobin content. It has few mitochondria, and its fibers valence The number of missing or extra electrons of an atom or a
are constructed of many clearly defined fibrils. These con- molecule.
tract rapidly and derive most of their energy from anaerobic van der Waals forces The close-ranging, relatively weak attraction
metabolism. exhibited between atoms and molecules with hydrophobic
2-deoxyribose 5-carbon sugar that forms a major structural com- properties.
ponent of DNA. van't Hoff equation An equation used to calculate the Q,, of a bi-
tympanic membrane The eardrum. ological function.
tympanum The middle ear cavity; houses the auditory ossicles. varicosities Swellings along the length of a vessel or fiber.
type J receptors Abbreviation for juxtapulmonary capillary re- vasa recta The capillary network that surrounds the loop of Henle
ceptors found in the lung that, when stimulated, elicit the sen- in the tubules of the mammalian kidney.
sation of breathlessness. vasa vasorum The tiny arteries and veins that supply nutrients and
remove waste products from the tissues in the walls of larger
ultradian rhythms Biological rhythms with a periodicity of greater blood vessels.
than a day in length. vas deferens A testicular duct that joins the excretory duct of the
ultrafiltrate The product of ultrafiltration. seminal vesicle to form the ejaculatory duct.
ultrafiltration The process of separating colloidal or molecular vasoactive intestinal peptide Peptide hormone regulating intesti-
particles by filtration, using suction or pressure, by means of nal phase of gastric secretion.
a colloidal filter or semipermeable membrane. vasoconstriction Contraction of circular muscle of arterioles, de-
ultraviolet light Light of wavelengths between 180 and 390 nm. creasing their volume and increasing the vascular resistance.
uniports Carrier proteins which transport a single solute from one vasodilation A widening of the lumen or interior space of the
side of the membrane to the other. blood vessels, increasing blood flow.
unitary currents Electrical currents due to the sudden opening of vasomotor Pertaining to the autonomic control of arteriolar
individual channels in the plasma membrane. constriction or dilation by contraction or relaxation of circu-
unit membrane The sandwich-like profile of biological membrane lar muscle.
seen in electron micrographs and believed to represent the vasopressin See antidiuretic hormone.
bimolecular leaflet with a hydrophobic center region between vasopressor A substance that induces arterial and capillary
hydrophilic surfaces. smooth-muscle contraction.
unsaturated In reference to fatty acid molecules, indicates that vector A carrier; an animal transferring an infection from host to
host. Also, a mathematical term for a quantity with direction, membrane potential across a membrane by means of feed-
magnitude, and sign. back control.
venous shunt A direct connection between arterioles and venules, voltage-gated ion channels Protein channels through the cell mem-
bypassing the capillary network. brane that allow ions to cross the membrane when they are
ventilation In respiratory physiology, the process of exchange of open. The conductance of these channels depends upon the
air between the lungs and the ambient air. transmembrane electrical potential difference.
ventral Toward the belly surface.
watt (W)A unit of electrical power; the work performed at 1joule
ventral horn The ventral part of gray matter in the vertebrate
(J)per second.
spinal cord in which motor neuron cell bodies are situated.
Weber-Fechner law Sensation increases arithmetically as a stimu-
ventral root A nerve trunk leaving the spinal cord near its ventral
lus increases geometrically; the least perceptible change in
surface; contains only motor axons.
stimulus intensity above any background bears a constant
ventricle A small cavity. Also, a chamber of the vertebrate heart.
proportion to the intensity of the background.
ventricular zone The region of the brain that surrounds the cere-
white matter Tissue of the central nervous system that consists
bral ventricles; in embryonic vertebrates, cells of the ventricu-
mainly of myelinated nerve fibers.
lar zone remain mitotic and generate the neurons of the brain
Wolffian ducts The embryonic ducts that are associated with the
and spinal cord.
primordial kidney and that become the excretory and repro-
venule A small vessel that connects a capillary bed with a vein.
ductive ducts in the mate.
vestibular apparatus The collection of vertebrate organs of equi-
work Force exerted upon an object over a distance; force times
librium in the inner ear.
distance.
villi Small, fingerlike projections of the intestinal epithelium.
viscosity A physical property of fluids that determines the ease X-ray diffraction The method of examining crystalline structure
with which layers of a fluid move past each other. using the pattern of scattered X rays.
visible light Light of wavelengths between 390 and 740 nm. Xylocaine The trade name for lidocaine, a local anesthetic related
visual cortex The cerebral cortex in the occipital region of the cere- to procaine.
brum; devoted to processing visual information.
visual streak (retinal streak) A retinal structure-found in the eyes Z disk (Z line, Z band) A narrow zone at either end of a muscle
of some species that inhabit plains-in which photoreceptors sarcomere, consisting of a latticework into which the actin
are packed into a horizontal streak across the retina, provid- thin filaments are anchored.
ing high resolution along the visual horizon. This pattern is zeitgeber Environmental factors that entrain biological rhythms.
similar to the fovea of primates, but it has a different shape. zero-order kinetics Kinetics in which the rate of the reaction is in-
vital capacity The maximum volume of air that can be inhaled into dependent of the concentration of any of its reactants. This
or exhaled from the lungs. would occur if the enzyme concentration were the limiting
vitalism The theory that postulates that biological processes can- factor.
not adequately be explained by physical and chemical zonula Zone.
processes and laws. zonula adherens Form of desmosome in epithelial cells that form
volt (V) MKS unit of electromotive force; the force required to a belt of cell-to-cell adhesion under tight junctions.
induce a 1ampere (A) current to flow through a 1 ohm (R) zonula occludens Tight junctions between epithelial cells, usually
resistance. having a ring-shaped configuration and serving to occlude
voltage (E or V) The electromotive force, or electric potential, ex- transepithelial extracellular passages.
pressed in volts. When the work required to move 1coulomb zwitterion A molecule carrying both negatively and positively ion-
(C)of charge from one point to a point of higher potential is ized or ionizable sites.
1 joule (J),or 114.184 calories (cal), the potential difference zygomatic cells See chief cells.
between these points is said to be 1 volt (V). zygote A fertilized ovum before first cleavage.
voltage clamping An electronic method of imposing a selected zymogen See proenzyme.
Page numbers in italics indicate illustrations.