Lecture Notes on General Histology

For
Medical Students

Naama Abdulgader, MD, PhD, Professor

Department of Histology and Medical Genetics
Faculty of Medicine
Tripoli University

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 CONTENTS

1. Introduction and Microscopy ………………… 3

2. Cytology and Cell Biology …………………… 22
3. Epithelial Tissue ……………………………... 61
4. Connective Tissue …………………………… 87
5. Blood ………………………………………… 115
6. Cartilage ……………………………………… 138
7. Bone ………………………………………….. 145
8. Muscles ……………………………………… 161
9. Nervous Tissue ……………………………… 178
10. Circulatory System ………………………... 205
11. Immune System and Lymphoid Organs …… 222
12. Integumentary System …………………….. 249

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 INTRODUCTION

Histology means the study of tissue components by the use of microscopes

(light and electron) and stained sections of tissues. The main objective in
studying histology is to identify the mammalian tissues quickly and
accurately.

The human body is composed of systems, which are made of organs. Organs
are made of tissues which are composed of cells. The cell size is measured by
special units and examined by microscopes.

The main idea in the study of histology is to know the general and specific
features of tissues and organs and memorize them very well. In addition to
understanding the histology and ultrastructure of cells and tissues, it is also
important to correlate the morphological features with an understanding of
the biochemistry and physiology of these structures.

Remember that histology is one of the most useful courses, because it is the
core subject in the study of function, macroscopic and morbid anatomy, and
cell and molecular biology. Moreover, most medical researchers depend on
histology in their research.

Units of measurements:
1 centimeter (cm) = 10 millimeters (mm)
1 millimeter (mm) = 1000 micrometers (um)

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1 micrometer (1um) = 0.001mm
1 nanometer (1nm) = 0.001um
1 Angstrom (1 Å) = 0.1nm = 0.0001um

MICROSCOPY
The basic type is the light microscope. Other types of microscopes are
modifications of the light microscope.

1. The Light Microscope (LM)

The light microscope is the most important machine in medical laboratory. It
uses a visible light source with a system of condenser lenses to send the light
through the object to be examined. The light microscope is used to enlarge
small objects and to reveal their fine details.
The light microscope is composed of:
1. Frame and mechanical parts
2. Optical (magnification) system
3. Illumination system

1. The frame supports the different parts of the optical system; it consists of
base, arm, stage with a central hole for the light to pass through and a body
tube carrying the optical system.

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2. The magnifying system is composed of:
 Eyepiece (ocular) with 8, 10, or 12 X of magnification.
 The objectives: four lenses with different magnifications:
3.5, 10, 40, 100 (oil immersion lens); they are carried on
an objective nose piece. Usually the oil immersion lense
is kept separately.
The objective lenses collect the light transmitted through the section from the
condenser, magnify the image and direct it to the ocular lens. The eyepiece
(ocular lens) magnifies the image and directs it to the viewer eye or to a
screen or a camera.
The property of making things to appear larger is called magnification.
The total magnification = ocular power x objective power
The property of disclosing the fine details is called resolution (R).
Resolution is the smallest distance between two particles that can be
distinguished from each other. Resolution is more important than
magnification, since it separates clearly between two points located close
together.
Resolution power of:
 Human naked eye: 0.1-0.2 mm
 LM is 0.1-0.2 um
 EM: 0.1-0.2 nm
The resolution (R) is directly proportional to the wavelength of light (‫ )גּ‬and
indirectly proportional to the numerical aperture (NA) of lenses. The NA of

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the lens is the sine of the angle of light entering between the middle and the
edge of the lens. Lenses with larger NA have better resolution.
Resolution (R) = constant x wavelength
Numerical aperture
R = 0.61 x ‫גּ‬
NA

The only way to improve the resolving power of a microscope (resolution)

is to reduce substantially the wavelength of the light. This was achieved
by the electromagnetic beam of the electron microscope.
3. The illumination system consists of: light source, condenser and iris
diaphragm to regulate the amount of light.

How to take care of your light microscope?

1. Do not touch lenses with fingers
2. Do not take objective lenses apart
3. Oculars should be cleaned with lens tissue
4. Carry the microscope by its arm with other hand under its base
5. Turn off the microscope and carefully cover it with its plastic cover.

2. The Electron Microscope (EM)

The Electron Microscope (EM) is better suited to study the details of cells
than LM. The detailed morphology revealed by EM may be called fine or
submicroscopic structures or ultrastructure. There are two types of the
electron microscopes, the transmission and scanning.

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A. Transmission Electron Microscope (TEM):
In this microscope, the ordinary light source is replaced by a beam of
electrons, which are emitted by heated tungsten gun or filament (cathode) in a
vacuum controlled system. There is a voltage difference between the cathode
and the anode which accelerates the electron beam and attracts the electrons
to the anode where they pass through its central hole. While passing in the
microscope tube, the electron beam is subjected to electric coils with
magnetic field, which deflect the electrons and change their path, thus called
electromagnetic lenses. The image is viewed directly on a fluorescent screen
because the human eye is not sensitive to electrons.

The tissue is treated with a double fixation method to preserve the

ultrastructural details. Buffered gluteraldehyde is used as a first fixative, and
then a special second fixative such as osmium tetra-oxide is used to stain
lipids and proteins. The tissue, then, embedded in resin or plastic in vacuum
oven. The resulting blocks are very hard; they are cut into very thin sections
(40-90nm) by the use of ultra-tome and a diamond or glass knife.
The thin sections are mounted on a special 100-400 mesh copper grids, and
stained with heavy metals such as lead and uranyl acetate.
The image is seen on a screen as a black and white and recorded on
photographs.
The electron microscope requires a vacuum-enclosed system, high voltage
(60-120KV), and mechanical stability.

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The high resolving power of EM made possible to study the details of the
interior of cells with a final magnification of 400,000 times.
The use of high voltage (500,000-1,000,000V) allowed the use of relatively
thicker sections to be examined to get 3-D images.

B. Scanning Electron Microscope (SEM):

This microscope provides a three dimensional image of the surface of fixed
and dehydrated tissues; it has less resolving power than TEM. The sample is
coated with gold, which emits secondary electrons after being hit with a
primary electron beam. The electrons do not pass through the specimen, but
scan sequentially the different surface points of the specimenand the resulting
image will be in black and white color.

Comparison between light (LM) and electron microscopes (EM)

Light Microscope Electron Microscope

Image Presented directly to Presented on a screen in shades of
the eye. Image keeps green. In photographs, image
the colors given by appears in grey scale or in black
staining. and white.
Magnification Up to X 1500 (times), High, up to X 400,000
wider field of sample (2,000,000?)
view; good
orientation.
Resolution 0.1 – 0.2 μm High, 0.1nm
Time By frozen section Tissue prcessing takes one day at

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processing sample can be least. Frozen tissue for
prepared in 20 min. cryofructure and histochemistry
Artefacts Many Fewer
Section 1 – 10 μm Very thin 40 – 90 nm
thickness
3-D image Can be constructed by Obtained by thicker section of
serial sections high-voltage EM, freeze-
fructured techniques, and SEM
Stain Routine H&E Heavy metals
Specimen size Can be large and alive Very small samples
Light source Visible light (electric) Beam of electron
Lenses Glass Electromagnetic

3. The Phase-Contrast Microscope

This type is used to study the living and unstained tissues. Its idea is based
upon the fact that light passing through the media of different refractive
indices changes direction and speed, thus creating contrast. It is equipped
with a lens system that enables it to convert the phase variation into intensity
variations, which are perceived as differences in brightness, and the unstained
object becomes visible.

4. The Interference Microscope

This microscope uses the same principle of the phase contrast microscope,
but with two beams of lights, which interfere with one another in a way to

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provide precise information on the density of cellular region even in the
living state.

5. Fluorescence Microscope
This microscope depends on the fact that certain substances present in nature
emit light of longer visible wavelength on exposure to ultraviolet or lase
light. This character is known as fluorescence and the molecules having this
character will appear colored or bright in a dark field. Some of these
substances are used to stain the cell components such as acridine orange
which stain specifically nucleic acids. With DNA, the acridine orange emit
yellowish green light and with RNA, acridine orange emits reddish orange
light. This microscope is used to localize nucleic acids in the cells, to localize
antigen-antibody complexes and to trace the pathway of nerve fibers.
6. The Polarizing Microscope
This type uses two filters, one of them is located below the condenser, called
the polarizer; the other filter is localized between the objective lens and the
eye piece and called the analyzer. When the axes of the two polarizers
become perpendicular on each other, no light passes resulting in a dark field
scene. This type is used to examine a crystalline substance or well ordered
fibrous molecule, such as collagen, microtubules and microfilaments which
appear light against the dark field. These substances alter the plane of the
entering polarized light and rotate the axis of the emerging light
(birefringence).

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 7. The Confocal Microscope
This microscope uses a very small beam of laser light for illumination, and a
highly computerized system. Its main principle is based on the fact that a
very small laser beam originating from one thin plane of the section passes
through a pinhole of a plate, while the rest of beams coming from other
planes are blocked by that plate. The small beam scans other planes of the
section, and thus able to collect serial optical sections from thick specimens.
Using filters to eliminate the unfocused images and the photomultiplier
detectors within a highly computerized system, made obtaining of a high
quality sharp three-dimensional image is feasible. It is used to optically
dissect the specimen and study the structure of biologic material after
fluorescent labeling of the area of interest, thus cannot be used as a routine
procedure.
Methods of study in histology
The human or animal tissue can be studied as: smears, spreads, sections or
teased material. Several methods are applied for this purpose, such as:
1. Microtechniques: using fixed, stained material, examined by light
microscope.
2. Electron microscopy: a fixed material is studied and high magnification
is obtained.
3. Autoradiography: A method used to study the tissue synthetic activities
by the use of radioactive isotopes by the use of light or electron microscopy.
Tissues or small animals are treated with radioactive isotopes of certain
molecules (precursor molecules) such as amino acids, nucleotides or sugars
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to synthesize larger molecules of proteins, nucleic acids or polysaccharides.
Tissue sections are prepared and covered with photographic emulsion
containing silver bromide and kept in the dark. When silver bromide is
exposed to radioactivity, it is transformed into metallic silver precipitating on
the sections as silver grains. Presence of silver grains and their number on
certain cells denote their synthetic activity and the path of their secretion.
4. Cell and tissue culture to study the living cells and tissues in vitro
(outside the body) and the effect of single molecule or substance on one type
of them. All the steps of culture are performed under strict sterile conditions.
Primary cell culture: Before cultivation, the cells should be dispersed
mechanically or by enzymes, and then suspended in nourishing medium
containing salts, amino acids, vitamins and serum components or spread out
on glass slides or petri dish. The resulting cell types are kept in vitro for
reuse and are known as a cell line. The cultured cells are genetically
programmed for a limited life span, but some of them show transformation
changes and become immortal, changing from normal to cancer cells.
5. Cell fractionation:
By this method, the cell components and organelles are separated
according to their sedimentation coefficient. This method is used to study
the chemical composition and function of the separated components in
vitro.
6. Histochemical and cytochemical techniques:
These methods are used to localize substances in tissue sections based on
the specific chemical reactions between macromolecules and production
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of insoluble colored substances that could be seen by the microscope (LM
or EM). Examples of the substances that can be studied:
a. Ions: such as calcium, iron, and phosphate.
b. Nucleic acids: DNA and RNA are studied by the use of basic dyes.
Feulgen reaction is specific to study and quantify the amount of DNA.
c. Proteins: to study the enzyme activity in the tissue by localising the
action of the enzyme with its substrate by producing colored insoluble
substance that can be seen by the microscope (LM or EM). The most
common enzymes to be studied:
 Phosphatases: acid phosphatase (lysosomes in macrophages) and
alkaline phosphatase (liver).
 Dehydrogenases: a group of enzymes that remove hydrogen from
one substrate to another that receives and precipitates it as an
insoluble colored substance e,g. succenic dehydrogenase
ofmitochondria.
 Peroxidase enzyme present in liver and kidney cells; it promotes
the oxidation of its substrates. It is used as a marker enzyme in
diagnosis of leukemia and other diseases.
d. Polysaccharides and oligosaccariades: the carbohydrates are present
either free (glycogen) or as combined with lipids as glycolipids or with
proteins as glycoproteins. The carbohydrate part can be demonstrated
by the use of PAS reaction.

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 PAS (Periodic Acid Schiff reagent): this method is used to stain
glycosamino-glycan [GAG], glycoproteins, and glycogen (salivary
amylase is used to break down glycogen), then counterstained by
hematoxylin.
e. Lipids: since lipids dissolve in routine preparations, it is better to
use dyes that dissolve in lipids. Frozen sections are dipped into
alcohol saturated with Sudan IV or Sudan black which stains lipid
droplets red or black respectively.
7. Freeze-fracture methods: Where the cell components are frozen to a
very low temperature to minus 170°C, then cleaved at the phospholipids
bilayer. The fractured cells are coated with carbon and platinum then
examined by EM.
8. Molecular high affinity interactions: these methods depend on specific
interaction between the molecules that have great affinity towards one
another. Labeled molecules with fluorescent compounds, enzymes, gold
particles or radioactive atoms are used to identify the specific interacted
molecules. Phalloidin (to demonstrate actin filaments), protein A
(immunoglobulins for staph aureus) and lectins (binds sugars in
glycoproteins, glycolipids, proteoglycans) are commonly used. These
methods are used to detect sugars, proteins and nucleic acids.
a. Immunocytochemistry: this is a highly specific reaction
between an antigen (foreign nonself protein) and its antibody
(immunoglobulin). Cells or tissues are incubated with
antibodies to their contents of proteins so that each antibody
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 binds specifically to its protein (antigen) denoting the protein
location in the cell that can be seen by microscope. Polyclonal
antibodies when the antibodies produced and collected contain a
mixture of several groups of antibodies for different parts of
proteins. Monoclonal antibodies when the antibodies for
different parts of proteins are collected separately. The methods
of immunocytochemistry could be direct or indirect.
The direct method is one step but less sensitive; the tissue
section is incubated directly with the antibody.
The indirect method is more sensitive, more expensive and
needs longer time. If rat protein (actin) is injected into another
animal of different species such as rabbit, the latter will produce
antibodies in his blood for rat actin (antirat actin antibodies).
Antibodies from a normal non-injected rabbit must be injected
into another animal of third species like a goat. Goat will
produce antibodies againt rabbit antibodies. Rat tissue sections
are incubated in solution containing antibodies produced by
rabbit against rat proteins, then washed and reincubated with
labeled goat antibodies against rabbit antibodies which already
recognized the rat protein.
b. Hybridization techniques: these techniques depend on binding
between two single strands of nucleic acids that recognize each
other if they are complementary. This method allows
identification nucleic acid sequences. In Situ Hybridization
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 when this technique is applied directly to cells, chromosomes or
tissue sections.
c. Polymerase chain reaction (PCR): This is an in vitro method
widely used in cell biology and diagnostic laboratories; it is used
to synthesize specific DNA sequences using two oligonucleotide
primers on both sides of DNA.

Tissue preparation for light microscopy

To prepare tissue for light microscopic examination, living or dead tissue can
be used. Living tissues such as tissue cultures in transparent dishes are
examined by phase-contrast microscopes. Samples of dead tissue are used in
the form of:
 Sections on glass slides
 Smears on glass slides for blood, mucus, urine, and bone marrow
 Spreads for areolar connective tissue
 Teased samples for muscle tissue
Microtechnique:
To prepare histology sections and get the tissue in its final shape on a
microscopic slide, the tissue must pass through the following steps:
1. Obtaining the tissue in small pieces to facilitate penetration of the fixative
into the tissue.
2. Fixation:

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Fixation could be done by physical (freezing) or chemical means
(formaldehyde and gluteraldehyde).
The tissue is immediately placed in a fixative such as buffered isotonic
solution of 4% formaldehyde for routine light microscopy.
The functions of a fixative are:
 To stabilizes or cross-links proteins, and protein conjugates for
firmer structures of the cell.
 To prevent autolysis (tissue digestion by enzymes present within
their cells).
 To preserve tissue and molecular structure.
Lipids, minerals, glucose, and smaller molecules are usually lost during
processing.
Intravascular perfusion of a fixative is very helpful to fix the whole body and
improve the quality of fixation.
3. Dehydration:
This means the removal of water from the tissue by using series of
increasing concentration of alcohol solutions, a mixture of ethanol and
water, from 70% to 100% ethanol.
4. Clearing:
Alcohol must be removed and replaced with a solvent agent that mixes
with molten paraffin such as xylene, toluene, and cedar wood oil.
Clearing agent makes the tissue clear and transparent.
5. Embedding or tissue impregnation:

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 Once the tissue is infiltrated with the solvent, it is placed in the
embedding agent in the oven until the tissue is thoroughly infiltrated by
the embedding agent that gives rigidity to the tissue.
Paraffin is used for embedding in routine light microscopy; paraffin melts
at certain temperature (58 – 60°C) as the heat helps the solvent to
evaporate and to be replaced by paraffin.
Paraffin is allowed to solidify at room temperature, in special frames to
give a solid block (containing the tissue) for sectioning.
Resin is an embedding agent for electron microscopy, but it can be used
also for light microscopy for thinner sections.
6. Sectioning (Section cutting) and mounting:
The tissue is then cut into thin sections 1 - 10 micrometers with a
microtome. The sections are then mounted on glass slides and stained.
Frozen sections are obtained after tissue fixation by freezing, then cut into
sections by cryostat (freezing microtome), mounted on glass slides and
stained. This method is used in hospitals for quick diagnosis during
surgery and to study the enzymes by histochemical methods.
7. Rehydration: by descending grades of alcohol.
8. Staining:
To stain the tissue and make some parts of it appears darker than the other
parts by the use of dyes which stain the tissue components selectively.
The dyes are either acidic or basic.
When tissue structures are stained with basic dyes such as hematoxylene,
methylene blue, and toluidine blue, they are said to be basophilic.
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 Routine staining of human material usually employs haematoxylin, which
reacts preferentially with nucleic acids and acidic groups of proteins. The
nucleic acids are concentrated in the nucleus and the rough endoplasmic
reticulum. A few cells have a lot of granular ER in their cytoplasm,
which gives a strong staining reaction, and the cell is said to be basophilic
(liking basic stains and haematoxylin).
Another dye is needed to stain the cytoplasm. Such a stain is eosin, acidic
in nature, and stains the cytoplasm of most cells red or pink. Some cells
stain bright red; they are said to be strongly eosinophilic or acidophilic
(liking an acidic stain). The cytoplasm may be basophilic, acidophilic or
neutrophilic.
Special stains are used to stain special structures such as glycogen
granules and fibrils; a counter stain is then used to reveal the nuclei and
the background of the special structures. Trichrome stains such as
Mallory and Masson`s stains are applied to differentiate between collagen
and smooth muscle fibers.
9. Dehydration by ascending grades of alcohol.
10. Clearing by xylene.
11. Mounting with cover slips.

Artefacts caused by tissue processing:

Faulty appearances in the microtechinques that can arise at all stages in
preparation of the section such as:
 Bad excision of the specimen from the body
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  Poor or inappropriate fixation
 Shrinkage or uneven shrinkage, leading to artificial spaces and
distorted relations
 Cutting marks from a bad microtome knife
 Bad mounting of the section, not flat on the slide
 Water, dirt or bubbles on or in the section
 Patchy or faded staining; unbalanced staining when more than one
stain has been applied
 Precipitate from the fixative or stain
 Tears and folds in the section.

The Most Common Stains in Light Microscopy

1. Hematoxylene and eosin (H & E):
This is the routine stain; it stains the nuclei blue and the cytoplasm pink.
2. Azocarmine:
This stain is used as a counter stain. It stains glycogen and mucigen.
4. Mallory-Azan:
It stains the nuclei red and the cytoplasm orange or blue.
5. Masson`s trichrome stain:
This stains the nuclei black or dark blue, the cytoplasm stains red, reticular
and collagen fibers stain blue.
5. Periodic acid-Schiff`s reagent and hematoxylene (PASH)

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This stains glycogen, mucin and carbohydrate containing tissue red or pink
and the nuclei blue.
6. Silver stain: Silver nitrate is used to stain reticular fibers, neurofibrils and
granules in enteroendocrine cells.
7. Orcein stain for elastic fibers.
8. Thionin stain: It is used to stain the Nissl`s granules blue in nerve cells.
9. Toluidine blue:
This stains the granules in mast cells reddish-purple, while the nuclei are
stained deep blue.
10. Trypan blue:
This stain is used as a vital to stain the living cells in vivo, or as a supra vital
stain to stain the living cells in vitro. In vivo, the stain is injected into the
living animals to demonstrate phagocytic cells which ingest stain granules.
11. Verhoff`s stain:
This is a specific stain for elastic fibers which stain black.
12. Osmic acid or osmium tetroxide:
This stain is used to stain the lipid contents of cells in black color e.g. myelin
sheath of nerve fibers.
13. Sudan III: stains lipids with orange color e.g. fat droplets in adipocytes.

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 THE CELL
The cell is the smallest structural and functional unit of all living organisms.
In brief the cell is the unit of life. The cell is not a static entity in life; its
chemical constitution and morphology are in continuous flux.
The cell is a mass of protoplasm surrounded by a cell membrane and contains
a nucleus and organelles.
Cells serving the same functions are grouped together and united by
intercellular substance to form tissue.
There are two types of cells: prokaryotes and eukaryotes. Compared to
prokaryotic cells, eukaryotic cells are larger and their nuclei are distinct,
surrounded by nuclear envelope to protect the genetic material, chromatin.
Eukaryotic chromatin is distinguished by having the basic proteins, histones
together with DNA in its basic units, nucleosomes.

Cell Differentiation
Cell differentiation means cell specialization where cells specialize in a way
to become able to perform one or more specific activity with great efficiency.
All the cells of an individual originate from a single cell, the zygote. The
first cell division of the zygote produces blastomeres which are totepotent i,e
able to differentiate into all cell types.

The process of differentiation is accompanied by morphological and chemical

changes such as developing special ability to synthesize specific substances.
The best examples of differentiation are the muscle cells which elongate and

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accumulate myofibrils containing contractile proteins in skeletal muscles, and
pancreatic acinar cells which synthesize digestive enzymes.

Differentiation is permanent and irreversible; cytokines are important

controlling factors for further differentiation.

Stem Cells

• A population of “reserve” quiescent cells which on stimulation,

undergo differentiation.

• One stem may produce several cell lines example: all blood cells are
derived from one stem cell.

Chemical composition of cells:

Although cells are different in shape and size, their basic chemical
composition is very much the same. They are composed of:
 Water is the main ingredient forming 75%
 Inorganic substances such as cat-ions, sodium and potassium
 Organic substances such as nucleoproteins, proteins, lipids and
complex carbohydrates.
Morphologically, the cell is composed of:
 Cytoplasm which is composed of matrix, organelles and inclusions.
 Nucleus

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 THE CYTOPLASMIC MATRIX
The cytoplasmic matrix, cytosol or ground substance, is the part of
cytoplasm intervening between the organelles and inclusions. It consists of
water, soluble enzymes, low molecular weight metabolites and ions.

THE CYTOPLASMIC INCLUSIONS

They are non-living (lifeless), temporary metabolic products of the cell, they
may or may not be enclosed by a membrane. They might be:
 Stored food as glycogen and lipids
 Pigments which might be:
a. Exogenous: carotene and carbon particles.
b. Endogenous: hemoglobin, hemosidrin, bilirubin, melanin and
lipofuscin.
 In pathological conditions: bacterial and viral inclusion bodies
may be found.

THE CYTOPLASMIC ORGANELLES

They are little organs of the cell, small, living, distinctive structures present
in almost all eukaryotic cells, and they have limiting membranes and perform
specific functions. The cell organelles are:
1. Endoplasmic reticulum (smooth & rough)
2. Golgi complex
3. Mitochondria
4. Lysosomes

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5. Peroxisomes (microbodies)
6. Proteasomes
7. Ribosomes
8. Centrioles
9. Microtubules
10. Microfilaments
11. Intermediate filaments

The Cell Membrane

It is a limiting membrane that surrounds the cytoplasm, and separates it from
the extracellular environment. However, the cell membrane contains
transmembrane integral proteins called integrins which link the cytoplasmic
cytoskeltal elements to the extracellular medium and allow constant cell-
environment interaction between the extracllular matrix and the cytoplasm.
By light microscope (LM), the cell membrane is not visible.
In EM, when stained with osmium, the plasma membrane appears as a tri-
laminar structure, made up of inner and outer electron-dense layers and an
intermediate electron-lucent layer. It is called a unit membrane because it is a
common structure to all cell membranes.
The membrane total thickness is 7.5 – 10 nm.
On the outer surface of the cell membrane, there is a layer of glycoproteins
and glycolipids. This layer is called the cell coat or fuzzy coat or glycocalyx.

Molecular structure of the cell membrane:

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Chemically, plasma membrane is made of lipids, proteins, and carbohydrates.
The arrangement of the membrane components is interpreted on the basis of
fluid mosaic model.

The carbohydrates:
These are present in the form of oligo-saccharides attached to proteins
forming glycoproteins, or attached to lipids forming glycolipids.
Carbohydrates have a role in the formation of glycocalyx and in cell to cell
recognition and interaction and in cellular adhesions.

The membrane lipids:

Its main structural elements are made of lipid bilayer of phospholipids (e.g.
lecithin and cephalin) and some cholesterol (increases the membrane
fluidity). Phospholipids are made of globular phosphate heads and tails. The
heads are polar, hydrophilic and arranged on the inner and outer sides of the
lipid bilayer. The tails are made of hydrocarbon double chains of fatty acids
which are non-polar, hydrophobic and directed towards the center of the lipid
bilayer.
The trilaminar appearance is due to deposition of the reduced osmium on the
areas of polar phosphate heads.
Phospholipids are not equally distributed on both sides of the membrane
leading to membrane asymmetry.

The membrane proteins:

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The proteins which make an important constituent of the cell membrane are
of two types according to their arrangement in the lipid bilayer.
1. The intrinsic (integral) membrane proteins which are tightly bound to the
lipid bilayer and could be extracted only by the use of strong drastic
measures such as detergent EDTA. They are partially or completely
embedded in the lipid bilayer. The proteins which are incorporated across
the whole thickness of the membrane are called transmembrane proteins
and they act as pump channels for water soluble substances. Integrins are
transmembrane integral proteins that link the cytoskeletal elements with
the extracellular environment allowing cell-environment interaction.
When the transmembrane proteins span the membrane once, it is
called one-pass transmembrane proteins; if they span the membrane
more than once, they are called multi-pass transmembrane integral
proteins.
2. The extrinsic (peripheral) membrane proteins which are loosely bound to
the membrane, and can be easily extracted by simple salt solutions.
The difference of protein composition on both sides of the lipid bilayer leads
to membrane asymmetry.
The proteins inside the lipid bilayer give the appearance of mosaic, hence the
name fluid mosaic model.
Under certain conditions, proteins move laterally inside the lipids of plasma
membrane and group in one place giving what is called capping phenomenon.

Functions of proteins in cell membrane:

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1. Act as structural units
2. Act as pump channels
3. Act as receptors
4. Act as enzymes
5. Act as adhesion molecules; examples of these molecules:
 Spectrin provides a fine skeleton just beneath the cell
membrane attached to cell membrane by ankyrin, helps to
control cell shape and movement.
 Cell adhesion molecules (CAMs)
 Integrins are cell surface molecules helping the cells to choose
to which extracellular molecules to bind to.
 Connexins are proteins which combine as hexamers to form
connexons or gap junction to allow passage of ions and small
molecules between the cells.
 Occludins are proteins which present in tight junctions to seal
and prevent passage of materials between epithelial cells.

Functions of the cell membrane:

1. Protects the cell contents
2. Firm attachment to other cells through membrane specialization.
3. Movement of the cell itself by pseudopodia
4. Movement of materials outside the cell by cilia in ciliated epithelium

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5. Acts as selective barrier controlling the passage of substances in and out
of the cell.
6. Active transport of certain ions using energy (sodium-potassium pump)
7. Carrying specific recognition sites for hormones, enzymes and antigen-
antibody complexes
8. Cell-environment interactions (transmembrane proteins integrins)
9. Endocytosis: uptake of macromolecules by the cell membrane
10. Exocytosis (emiocytosis): releasing the material of secretory products to
outside the cell.
11. It conducts the wave of excitation in nerve and muscle cells.

Cell Membrane Transport

All the materials the cell gets from or sends to the environment has to go
through the cell membrane. It is basically a phospholipid bi-layer with
various proteins embedded into it. The cell membrane is semi-permeable that
is some materials can get through it (water, carbon dioxide, oxygen, very
small polar molecules such as ammonia, and cholesterol); while others can't
such as hydrogen ions, glucose, amino acids and macromolecules.

Transport mechanisms: (passive and active)

 Passive transport mechanisms do not require energy. By these

mechanisms, water, molecules, and ions move in and out of the cells

29
 freely from areas of high concentration to the areas of low
concentration.

Facilitated transport is considered as passive duffusion since it does

not require ATP. It is termed facilitated because the channel proteins
facilitate or help molecules move through the cell membrane.

 Active transport mechanisms: the molecules move from a region of

lower concentration to a region of higher concentration. As this
process does not naturally occur, the cell has to use energy in the form
of ATP to make active transport to bring material into the cell. The
"bulk" flow of endocytosis and exocytosis enables the cell to take in
very large packages of molecules, a process that requires ATP.
Endocytosis and Exocytosis

Endocytosis is the uptake of material by the plasma membrane; by

invagination, and formation of small vesicles or large vacuoles.
a. Pinocytosis is the uptake of extra-cellular fluid by the cell
membrane.
b. Phagocytosis is the uptake of solid matter.
Exocytosis is the release of material out of the cell through the cell
membrane.

Pinocytosis:

30
1. Fluid-phase pinocytosis: small invagination of the cell membrane to
surround the extracellular fluid forming pinocytotic vesicles which fuse with
the primary lysosomes producing secondary lysosomes. In endothelial cells
of capillaries, pinocytotic vesicles are used for transcapillary transport.
Pinocytotic vesicles move to the opposite side of the cell where they fuse
with the cell membrane to empty their contents out of the cell.
2. Receptor-mediated pinocytosis: this is a highly selective process of
uptake of specific proteins like hormones, growth factor and
cholesterol.
Receptors are brought to the plasma membrane by vesicles from the trans
region of the Golgi complex. These receptors are scattered over the cell
surface, or move laterally in the membrane and collect in specialized regions
called clathrin coated pits. When the ligand binds to its specific receptor, the
ligand-receptor complexes accumulate in the coated pits.
On the cytoplasmic side of cell membrane and opposite the coated pits, lies a
special coat made of several proteins, but mostly of clathrin. Clathrin
molecules are arranged as pentagons and hexagons. The coated pits
invaginate and pinch off forming coated vesicles which carry the ligands with
their receptors. Inside the cytoplasm, these vesicles soon loose their clathrin
coat to become uncoated and fuse with early endosomes to form larger
vesicles (receptosomes).
All endosomes are usually acidic due to the action of a proton-pumping
ATPase of their membranes. Acid pH helps the ligands to leave their
receptors to return to cell surface to be reused. Ligands are then, transferred
31
to late endosomes which join primary lysosomes to give rise to secondary
lysosomes which start the process of digestion or degradation. A good
example is LDL which is taken into the cell by binding to its LDL receptor at
the cell surface. On reaching the early endosomes, LDL dissociates from its
receptors, and the receptors are taken back to the cell surface for reuse. The
LDL remains in the endosome and is delivered to lysosomes for processing;
LDL is the primary carrier of cholesterol in the blood. In some people, the
LDL receptors are defective, so uptake of cholesterol-containing LDL from
the blood into liver cells is reduced and LDL-cholesterol accumulates in the
blood resultin in hypercholesterolemia. Another example is the iron-transport
protein transferrin which binds receptor (transferrin receptor) and is
transported to cytoplasm by the coated vesicle which soon loses its coat and
joins endosome. In the acidic medium of endosome, the iron is released from
transferrin, and the transferrin-receptor complex is returned to cell surface to
be re-used.

Endosomes:
Endosomes are membranous compartments inside eukaryotic cells to
transport molecules in endocytic vesicles from plasma membranes to
lysosomes. When endosomes are closer to the surface, they are called early
endosomes, which mature and move deeper into the cytoplasm where they
are called late endosomes. Some authors refer to late endosomes as
multivesicular bodies; meanwhile others consider multivesicular bodies as a
variant of secondary lysosomes.
32
Phagocytosis:
Phagocytosis means ingestion of solid matters, or cell eating. Cells
with phagocytic activity, like neutrophils and macropages engulf and
remove foreign bodies and microorganism by forming cytoplasmic
processes (pseudopodia) which surround the foreign body or
bacterium and draw it inside the cytoplasm forming phagosomes or
phagocytic vacuoles.
Exocytosis:
It is the fusion of the vesicles or membrane-bound granules with the cell
membrane to release their contents into the extracellular space, e.g.
pancreatic acinar cells. Exocytosis is controlled by the level of calcium ions
in the cytosol.
Lysosomes
Lysosomes are membrane limited vesicles, 0.05 - 0.5 um in diameter. Only
the large ones can be resolved by LM. They are enclosed by a single
membrane and contain hydrolytic enzymes which are active at acid pH.
Lysosomes present in large number in cells with phagocytic activity such as:
macrophages, neutrophils and osteoclasts.
Lysosomal enzymes are synthesized in RER then transferred to Golgi
complex, where they are concentrated, then packaged as primary lysosomes.
The small primary lysosomes can be detected only by histochemical methods.
There are two types of lysosomes, primary and secondary.
A. Primary lysosomes: they are small about 0.05 um in diameter,
homogenous and do not contain digested materials.
33
B. Secondary lysosomes: are primary lysosomes plus phagosomes. They are
larger, heterogenous and contain digested materials.
Function:
Lysosomes are responsible for intracellular digestion and turnover of
cytoplasmic organelles.
After digestion, nutrients diffuse to the cytoplasm; the undigested material
which stays in lysosomes is called residual bodies. Large number of
residual bodies accumulates by age and known as age pigments containing
lipofuscin or lipochrome pigments which are detected in nerve cells, liver
cells and cardiac muscle cells.
Heterophagy is the process of digestion of materials from the surrounding
environment outside the cell.
Autophagy is the process of digestion of materials from inside the cell such
as aged cellular organelles, resulting in autophagosomes.
Normally, lysosomes are intact and their hydrolytic enzymes do not have
access to the intracellular structures. Under pathological conditions or lack of
oxygen, lysosomes rupture and their enzymes escape leading to self-
destruction of the cell or cell lysis or autolysis.
Disorder of lysosomal enzymes: (lysosome storage disease)
 In lysosomal deficiency diseases, the inherited absence of an enzyme
causes the massive accumulation of the material which is normally
broken down, such as:
 Glycogen storage disorder: from a lack of a-glucosidase enzyme.

34
  Hurler's disease: a devastating disorder, where a deficiency in
lysosomal a-L-iduronidase causes intra- and extra-cellular excess
accumulations of dermatan-sulphate and heparan-sulphate
glycosaminoglycans. Patients will have dwarfism, mental retardation,
and many cardiovascular defects which cause early death.

Peroxisomes (Microbodies)
They are spherical membrane bound organelles present in large number in
liver and kidney cells. They contain oxidative enzymes such as catalases and
oxidases.
Peroxisomes use oxygen but they don’t produce ATP and have no role in cell
metabolism. Oxidases form hydrogen peroxide which is very harmfull to the
cells. Hydrogen peroxide will be immediately decomposed by catalases to
water and oxygen and thus protecting the cells from hydrogen peroxide
irreversible cellular damage.
Functions:
 Peroxisomes have a role in purine degradation
 Involved in lipid metabolism and beta oxidation of long-chain fatty
acids
 Bile acid and cholesterol formation.
 Produce hydrogen peroxide and convert it to water
 Degradation of toxic molecules in prescription drugs and ingested
ethyl alcohol.

35
Clinical Notes:
The congenital absence of peroxisomes in the cells of brain, liver and kidney
causes dysfunction of these organs and produces a severe fatal genetic
disorder, Zellweger's syndrome (cerebrohepatorenal syndrome).

Proteasomes
Proteasomes are complexes made of multiples of proteases that digest and
degrade proteins. This process is important to remove the excess of enzymes
and the unnecessary, faulty defective proteins by conjugation to ubiquitin to
be taken to proteasomes for degradation.
Ubiquitin is a small regulatory protein present in eukaryotic cell as free or
attached to other proteins. The most prominent function of ubiquitin is
labeling proteins for proteasomal degradation.

Endoplasmic reticulum
It is a membranous organelle, composed of tubules and cisternae, branching
and anatomozing giving the appearance of a network.
It is divided into two types:
 Smooth endoplasmic reticulum (SER) with no ribosomes
 Rough endoplasmic reticulum (RER) with ribosomes.

36
A. Rough endoplasmic reticulum (RER), it is called rough due to
presence of ribosomes attached to outer surface of its cisternae. Its
membranes are continuous with the outer layer of nuclear envelope.
It is highly advanced in cells specialized in protein secretion, the so called
protein secreting cells such as:
1. Pancreatic acinar cells which secrete digestive enzymes
2. Fibroblasts which secrete collagen fibers to help wound healing by scar
formation
3. Plasma cells which secret immunoglobulins or antibodies.
Cells rich in RER appear basophilic and granular in LM.
RER has attachment site for the large subunits of ribosomes due to presence
of ribophorin I and ribophorin II.
Ribophorin I and II are integral membrane proteins which present in the
membranes of RER; they provide attachment sites for the large subunits of
ribosomes as well as channels for the newly synthesized protein to enter the
lumen of RER.
Functions of RER:
1. Mainly, to segregate the newly synthesized protein designated for export
2. Limited proteolysis
3. Initial glycosylation
4. Assembly of multichain protein
5. Phosphorylation of phospholipids

37
All protein synthesis in cells starts on free polysomes; the mRNA of
proteins designated for export has a signal sequence at its 5` end of the
molecule. The signal sequence interacts with a signal recognition particle
(SRP) which stops further poypeptide elongation. (SRP = 6 polypeptides
+ RNA). When SRP-polysome binds receptors on RER called docking
proteins, SRP is removed by signal peptidase enzyme and protein
synthesis continues. Destination of synthesized proteins by RER:
 Storage inside the cells (lysosomal enzymes)
 Export, pancreatic and endocrine cells
 Used in cell membranes as integral membrane proteins
B. Smooth endoplsmic reticulum (SER):
It is composed of a network of layers of flattened sacs (cisternae) connected
by tubules. It differs from RER, in having no ribosomes and its cisternae are
more tubular. SER is continuous with RER; it is abundant in liver cells,
steroid secreting cells, and parietals cells of the stomach and muscle cells.
Functions:
1. Detoxification of noxious substances or drugs as barbiturates by
hydrolysis, conjugation or oxidation (liver)
2. Glycogen mobilization and breaking it down by the action of glucose-6-
phosphatase (liver)
3. Synthesis of plasma lipoproteins (liver).

38
4. Synthesis of phospholipids to all membranes and steroid synthesis (adrenal
cortex, testes, ovaries). Phospholipids are transferred along the cytoskeleta
elements, or by direct continuity with RER or by protein carriers.
5. Ca sequestration, in muscle where SER is referred to as sarcoplasmic
reticulum. It synchronizes contractions and relaxations throughout the fibre,
by moving calcium ions to and from the sarcoplasm.
6. Involved in secretion of chloride ions as in parietal cells of gastric glands

Ribosomes
They are small electron-dense particles, found in all eukaryotic cells, they
cannot be seen by LM, and only basophilia is seen due to content of RNAs.
Composition: They are composed of rRNA plus proteins. Ribosomal RNA
is made up in nucleolus.
The protein is synthesized in cytoplasm; and then enters the nucleus through
the nuclear pores to associate with rRNA to make ribosomes which enter the
cytoplasm through nuclear pores to start protein synthesis.
Ribosomes are composed of two subunits, small and large attached by
mRNA.
Aggregates of ribosomes are called polyribosomes or polysomes.
Ribosomes are present as free or attached to RER.

Functions of ribosomes:
 Free ribosomes: synthesize proteins for local intracellular use

39
  Attached ribosomes: synthesize proteins for export and integral
membrane proteins.
Ribosomes are highly basophilic due to presence of phosphate groups in
rRNA which react with the basic stains as hematoxylene giving intense
basophilia to cytoplasm in LM. The basophilia is called ergastoplasm or
Nissl bodies which are best seen in neurons.

Golgi complex
It was named after Golgi who found it first in nerve cells with silver
impregnation method, in LM, as a tangled network. With routine
haematoxylin and eosin staining in certain cells, the juxta-nuclear vacuole
reveals the site of the Golgi structure as a pale negative image. There may be
more than one Golgi apparatus in large active cells.
Golgi apparatus is composed of a complex of stacked smooth cisternae-
enclosing lamellae like saucers, tubules, and vesicles of various sizes:
 Flattened cisternae
 Transfer small vesicles are present on the convex cis face.
 Large condensing vacuoles are present on the concave, trans,
maturing face.
Golgi complex lies between the nucleus and the apex of the cell
(supranuclear), close to RER. It has:
 Convex, forming, cis or immature face which is osmiophilic
 Concave, maturing, trans-face, argyrophilic.

40
Functions of Golgi complex:
1. Concentration and packaging of secretory proteins and hydrolytic
enzymes (lysosomes)
2. Glycosylation (adding oligosaccharides to proteins or lipids), and
completing the synthesis of complex sugars.
3. Sulfation (adding sulfate groups as in sulfated proteoglycans in cartilage).
4. Clipping and chemical modification of protein pro-molecules for
activation, such as pro-insulin is activated into insulin, where pro-insulin
is clipped in the middle and the two ends are associated together with a
disulfide bond giving rise to active insulin. Failure of this process leads to
diabetes mellitus.

Protein synthesis:

1. Ribosomal RNA and ribosomes are synthesized in the nucleolus.

2. In the cytoplasm, ribosomes are held in clusters by mRNA.
3. Protein synthesis begins on free ribosomes (polysomes) and it is
controlled by messenger RNA (mRNA).
4. Messenger RNA (mRNA) is synthesized by transcription of nuclear
DNA, and leaves the nucleus to cytoplasm through the nuclear pores.
5. The proteins destined for export have an additional sequence called
signal sequence which interacts with SRP (signsl recognition particle)
to prevent further elongation of polypeptide chain.

41
 6. Polypeptide elongation is allowed after SRP-polysome complex binds
the docking protein (a receptor on RER membrane) and the SRP is
released. Also the signal sequence is removed by an enzyme present
on the inner surface of RER called signal peptidase.
7. Transfer RNA (tRNA) brings activated amino acids to the ribosomes
for linking together, thus producing specific a polypeptide chain or
protein as specified by the transcribed nuclear DNA.
8. Inside RER, structural as well as chemical modifications take place
like: hydroxylation, glucosylation, sulfation and phosphorylation.

Fate or distination of proteins synthesized in RER:

 Intracellular storage in lysosome

 Provisional intracellular storage for export as needed (pancreatic
acinar cells)
 To be used as integral proteins in membranes.

Mitochondria
Mitochondria are granular or filamentous cell organelles that can be seen by
LM, they are 0.5 - 1 um wide and 1 - 10 um long. They are the power house
of the cells, their number increases in more active cells such as cardiac
muscle, renal tubules and liver cells. They have self-duplicating ability
whenever needed where their number might reach up to 2500 in a liver cell.
Mitochondria are composed mainly of proteins, small amounts of lipids,
DNA and RNA.
42
Functions of mitochondria:
 Generation of energy to support the various forms of cellular
activities; they are the site of cellular respiration.
 Control the Ca++ level inside the cell.
Mitochondria are found in large number in places where energy is needed
such as: apical end of the ciliated cells, bases of ion transport cells and in the
middle piece of sperm.
Ultrastructure of mitochondria:
By EM, mitochondria are surrounded by two membranes: An outer smooth
mitochondrial membrane and inner folded making cristae. Between the
inner and outer, there is an intramembranous space. Inside the cristae, there
is an intracristal space, which is continuous with the intramembranous space.
Inside mitochondria, the intercristal space contains the matrix.
The matrix contains enzymes for lipids and protein synthesis, enzymes for
citric acid cycle, calcium ions binding sites, circular DNA coding for 13
respiratory-chain proteins, RNA and ribonucleoproteins.
Mitochondria originate either from pre-existing ones by growing then
dividing by fission, or from prokaryotic cells that adapted and lived in
eukaryotic cells by symbiosis.
Centrioles
They are also called division bodies or centriolar complex; two centrioles
form a centrosome.

43
The centrioles are small cylindrical structures about 0.15 x 0.3 um; there are
two centrioles for each cell, perpendicular on each other. Centrioles are seen
with difficulty by LM as dots.
By EM, each centriole appears to be composed of nine triplets of
microtubules circularly arranged.
Centrioles have self-replicating ability; they duplicate themselves in the S
phase of cell cycle. Centrioles are important in cell division; during cell
division, each pair of centrioles moves to one pole of the cell and become the
organizing centers of microtubules in mitotic spindles.

CYTOSKELETAL (FILAMENTO-TUBULAR) SYSTEM

Cytoskeleton is composed of:
 Microtubules
 Actin-myosin filaments (thin and thick)
 Intermediate filaments
General functions of cytoskeleton:
1. Structural proteins
2. Provide form and shape of cell
3. Involved in cytoplasmic and cellular movement
4. Dynamic organizers of cell components and cell polarity
Microtubules
Microtubules are hollow non-branching tubes or cylinders 24 nm in diameter.
They are distributed throughout the cytoplasm. The microtubules are

44
composed of structural protein subunits called tubulin, made of alpha and
beta-tubulin heterodimers. They grow at one end called nucleation site by
polymerization of tubulin subunits.
Microtubules have variable length in different cells. The cross section of
microtubules consists of 13 protofilaments of tubulin.
Microtubules dissociate into their subunits by antimitotic drugs such as
colchicine and vinblastine. Their growth is under the control of microtubules
organizing centers (MTOC), which include basal bodies, cilia and
centrosomes.
Functions of microtubules:
1. Microtubules act as structural proteins
2. They have a role in intracellular transport of organelles and vesicles; this
process is controlled by special motor proteins (dynein and kinesin)
which use energy for moving molecules and vesicles along the
microtubules.
3. They provide the cell form and shape
4. They help to create cell polarity by moving certain materials to regions of
the membrane.
5. They form the bases of centrioles, basal bodies, cilia and flagella.
6. Form mitotic spindle and help separation of chromosomes in cell division.

Microtubules might be present as twos and called doublets, or as threes and

called triplets. An axoneme is comopsed of 9 doublets plus two central
microtubules, a structure known as (9 + 2). Each doublet in axoneme is
45
composed of two microtubules A and B, where A is a complete one, and B is
a C-shape, sharing with A two to three protofilaments.
Dynein arms are carried on A microtubule, they carry ATPase enzyme which
provides energy for microtubule sliding movement. Axoneme forms the
basic structure of cilia and flagella.
Cilia are shorter (2 - 3 um) than flagella (around 100 um), and cilia number is
greater while only one flagellum is present per cell, a good example is
spermatozoan.
A pathological condition affecting males called Kartagener`s syndrome or
immotile cilia syndrome, where the patient suffers from chronic chest
problems and infertility.
Filaments
According to size, filaments are subdivided into 3 types:
 Thick filaments (myosin): 15 nm in diameter
 Thin filaments (actin): 5 nm in daimeter
 Intermediate filaments: 10 nm in diameter

Intermediate filaments
Intermediate filaments are 10 – 12 nm in diameter, found in different cells,
and called different names such as:
1. In epithelial cells, they are called keratins where they function as
defensive and protective.
2. In mesenchymal cells where they are called vimentin

46
3. They are called desmin (skeletin) in smooth muscle and Z line of
skeletal and cardiac muscles
4. In glial cells, they are called glial filaments
5. In neuron, they are called neurofilaments
6. Nestin present is stem cells and serves as a stem-cell marker
Microfilamets or thin filaments
They are 5 – 7 nm in diameter, composed of globular protein actin, which are
arranged as double stranded helix. Microfilaments are fine threads visible in
EM, but may be aggregated into thicker fibrils visible in LM; they are under
the control of Ca ion concentration and cAMP.
Functions:
1. Microfilaments are involved in muscle contraction where they are present
in between thick myosin filaments (15 nm in diameter).
2. They are present just beneath plasmamembrane forming a thin sheath
called the cell cortex; also scattered in cytoplasm giving the cell its form
and shape.
3. In absorptive cells, they form a major component of the apical terminal
web and the core of microvilli.
4. They are associated with some cell structures such as secretory granules
and vesicles in endocytosis and exocytosis.
5. They form constriction in anaphase of mitosis and cell cleavage
6. Actin filaments form a major component of the apical terminal web
inserting into junctional complexes and running up inside the microvilli.

47
 THE CELL NUCLEUS
The cell nucleus is the part of the cell surrounded by a double membrane
envelope. It is structurally and functionally specialized in the storage and
expression of genomes. Its size varies from 1 - 19 um, its size is proportional
to the cell's metabolic activities. The nuclei of a specific normal tissue are
usually similar and uniform, while the nuclei of infected or malignant cells
have highly irregular nuclei with variable sizes and abnormal chromatin
morphology.
The number of nuclei per cell is usually one. It is said to be binucleated
when the cell has two nuclei, or a cell is said to be multinucleated when it has
several nuclei. An erythrocyte has no nucleus.
The nuclear components are:
1. Nuclear envelope
2. Nucleolus
3. Nuclear matrix, nucleoplasm or karyoplasm
4. Chromatin

1. Nuclear envelope
Nuclear envelope consists of two membranes, inner and outer nuclear
membranes, containing a narrow space called perinuclear cistern or space.
Both membranes are unit membranes.
The outer nuclear membrane has ribosomes on its outer surface and is
continuous with RER; the proteins synthesized on it are segregated in the

48
perinuclear cistern. The nuclear envelope is impermeable to ions and all
molecules.
Just beneath the inner surface of inner nuclear membrane lies the fibrous
lamina which does not block the nuclear pores. Both membranes are fused at
the nuclear pore complex.
The pore complex forms a controlled channel system between the cytoplasm
and the nucleus. The entire pore complex is made of more than 100 proteins;
it is 100 nm in diameter; the internal diameter is 40 - 60 nm. Nuclear pores
are octagonal having eight transverse fibers and eight radial fibers. The
number of pores depends on the cell type.
Nuclear pores permit ions and molecules with a diameter less than 9 nm to
pass freely without consuming energy.
Molecules larger than 9 nm pass through the pore complexes by the active
process through two steps. The first step without using energy when the
nuclear signal attaches to cytosolic protein forming a complex molecule
temporarly attaches to nuclear pore complex. In the second step, the protein
with nuclear signal is transported to nucleas by the use of energy leaving the
cytosolic protein in the cytoplasm.

2. Nucleolus
Nucleolus is round, dense, well-defined structure, it has no limiting
membrane, but heterochromatin is usually associated with the nucleolus.
There are 1-4 nucleoli per cell nucleus; more active cells have more nucleoli.
Nucleoli are rich in rRNA and proteins, they vanish during cell division.
49
By EM, nucleolus consists of:
i. Nucleolar organizer DNA, also called pars amorpha, it contains the
sequence of bases coding for rRNA. Five chromosome pairs of human
genome contain DNA nucleolar organizers.
ii. Pars fibrosa which is made of densely packed ribonucleoprotein fibers;
also contains the transcripts of rRNA genes.
iii. Pars granulosa which contains maturing ribosomes (15 – 20 nm).

Functions of nucleolus:
Nucleolus is the site synthesis of rRNA and assembly of ribosomal subunits.

3. Nuclear matrix
Nuclear matrix, karyoplasms, or nucleoplasm, is a semifluid substance
occupying the space between chromatin particles and nucleolus. It consists
of nucleoskeleton, fibrogranular ribonucleoprotein network, metabolites, ions
and fibrous lamina.
Fibrous lamina lies immediately beneath and connected to inner nuclear
membrane, it consists of three proteins: lamin A, B and C (72, 68, 62 kd).
The nucleoskeleton serves as attachment sites for DNA loops.

4. Chromatin
Chromatin is the genetic material contained in chromosomes, each of which
bears a large number of functional units called genes.

50
In interphase nucleus (non-dividing), the individual chromosomes are not
visible, only chromatin is seen.
Morphologically, chromatin as seen by the microscope is present in two
forms:
A. Euchromatin, which is dispersed, extended or diffuse, lightly stained
and highly active for transcription and replication. When the nucleus has
a large amount of euchromatin, it is said to be vesicular or open-face.
B. Heterochromatin appears as dense, tightly packed and darkly stained.
It is inactive for transcription and replication. Heterochromatin might
be:
 Constitutive heterochromatin which is condensed and inactive all the
time regardless of the state of the cell activity such as centromere.
 Facultative heterochromatin which might be condensed and inactive or
decondensed and active depending on cellular needs such as X
chromosome.

When a nucleus has large amount of heterochromatin, it is called massive or

closed-face nucleus.
During metaphase of mitosis, individual chromosomes become visible.
There are 46 chromosomes in humans (23 pairs), 22 pairs are called
autosomes (somatic chromosomes), and one pair is called sex chromosomes
or genosomes.

51
A male has 44 autosomes plus XY sex chromosomes. A female has 44
autosomes plus XX sex chromosomes. The autosomes are similar in both
males and females.
In females, there is an inactive X chromosome which is called Barr body or
sex chromatin.

Barr body or sex chromatin:

It is the dark staining heterochromatic mass lying against the inner surface of
the inner nuclear membrane of interphase female cells only. Barr body is
tested by the use of buccal smear which is said to be positive if 30% or more
of the cells have sex chromatin body. It is unreliable to test for sex using
buccal smear. Karyotyping should be considered. Barr body is believed to
represent the inactivated X chromosome.[ Lyon hypothesis: all of the X
chromosomes except one in a cell are inactivated early in development].
Barr bodies could be 2, 3, or more depending on the number of X
chromosome minus 1 for the active X chromosome. A female with
chromosome number 47, XXX has two sex chromatins, and an individual
with 49, XXXXY is having three Barr bodies.

Biochemistry of chromatin:
Chromatin is composed mainly of proteins, DNA and some RNA.
Chromatin has two major classes of proteins: histones and non-histones.

52
i. Basic proteins or histones, which are positively charged, rich in lysine and
argenine. There are five major fractions of histones which are: H1 (linker
histone), and the core histones: H2A, H2B, H3 and H4.
ii. Non-histone or acidic proteins are the least understood of chromosomal
components. They are a complex group of about 20 - 40 minor proteins,
they act as enzymes, structural units or genetic regulators.
All DNA in nucleus is packaged with proteins. The complex of nine histones
and DNA form nucleosomes; these are arranged as beads on a string forming
a 10 nm fiber. Nucleosomes are the basic repeating units of eukaryotic
chromatin. Each nucleosome consists of a coiled length of double stranded
DNA, about 166 base pairs together with an octameric complex formed of
two copies of each of the core histones (H2A, H2B, H3, and H4).
Each nucleosome has only one copy of H1 at the entrance and exit of DNA
around the core histones.
Higher order structure of chromosomes is obtained by interpretation of two
models. One of them is the solenoid model which proposes that the 10 nm
fiber supercoils into 30 nm fibers, then to 700 nm and finally to a
chromosome of 1400 nm. The other model is the superbead model which
proposes that 6 - 10 nucleosomes aggregate to form superbeads connected by
10 nm fiber which then condense to give 30 nm fibers.

The Cell Cycle and Cell Division

The cell cycle consists of an interphase (G1, S, G2) and mitosis (m).
The interphase is the phase between two mitotic divisions.
53
The stages of cell cycle:
1. G1 (Gap1): it is the phase after mitosis and before DNA synthesis
(presynthesis). It is the longest period of the cycle (25 hours) during which
RNA and proteins are synthesized in large amount including the proteins
which controll the cell cycle and cell volume. Cells that no longer divide like
nerve and muscel, enter the G0 phase.
2. S phase: (8 hours)
This is the interval of DNA synthesis, during which DNA becomes doubled
(4N in somatic cell and 2N in gametes) and duplication of the centrioles.
Before this stage each chromosome consists of a single chromatid, and by the
end of this stage, each chromosome consists of two sister chromatids joined
by a centromere. It takes about 8 hours.
3. G2 (Gap 2): It is the interval after DNA synthesis (post-DNA duplication)
and before mitosis; around 4 hours duration. During this phase, energy
accomulated, tubulin and chromosomal nonhistone proteins are also
synthesized. The maturation promotin factor (MPF), a complex protein
which induces the events of cell division is also synthesized in this pahse.
4. Mitosis (m): it is the shortest phase of the cycle (about one hour). It
includes cytokinesis (cytoplasm division), and karyokinesis (nuclear
division).
Regulation of the cell cycle:
The cell cycle check points are used by the cell to monitor and regulate the
progress of the cell cycle.
 Growth factors in low concentration
54
  Detection and repair of damaged DNA before replication at the
checkpoint G1 before transition to S phase.
 Prevent passage of the replicated damaged DNA at the check point at
G2 before the cell moves to mitosis.
 The protein p ⁵³ is responsible for the control mechanisms on the
previous check points at G1 and G2 transitions. This control protein
was found to be mutated in human cancer, thus allowing transfer of
damaged DNA to daughter cells and increasing cancer frequency.
MITOSIS
It is the process by which the somatic cell can divide to produce two daughter
cells identical in chromosome number and amount of genetic material (DNA)
to the parent cell. Mitosis is divided into 4 stages:
A. Prophase:
The chromatin, or uncoiled chromosomes condense and become more visible
under the light microscope as discrete long individual chromosomes.
Nuclear envelope breaks down by the addition of phosphate groups to the
proteins of fibrous lamina, then disappears. Each centrosome migrates to one
pole of the cell, the mitotic spindle is formed and the nucleolus disappears.
B. Metaphase:
Chromosomes become shorter and thicker and move to the center of the cell
to become attached to mitotic spindle fibers by the centromere at the cell
midline. Each chromosome appears to consist of two sister chromatids held
together by a centromere.

55
The site where the microtubule is inserted into the centromeric region is
called kinetochore, an electron-dense plaque made of DNA and protein .
C. Anaphase:
The centromere of each chromosome divides longitudinally, and each sister
chromatid (daughter chromosome) separates, and moves to one pole of the
cell by pulling action of microtubules.
D. Telophase: (nuclei of new cells reappear)
Daughter chromosomes reach the poles of cells and begin to decondense.
The nuclear envelope and nucleolus reappear, and two daughter cells are
formed, simultaneously, constriction of the cytoplasm caused by actin and
myosin filaments accomulated just beneath the cell membrane and division of
the cytoplasm occurs.
The daughter cells enter the interphase where the chromosomes loose their
discreteness and become chromatin.
Nerve and muscle tissue cannot regenerate because they do not divide after
birth.
MEIOSIS
It is the type of cell division which produces the haploid gametes (daughter
cells) from a diploid parent cell. The essential characteristics of meiosis are:
pairing, crossing over, and reduction of chromosome number.
Meiosis is divided into :
 Meiosis I or reduction division.
 Meiosis II or equational division.

56
 MEIOSIS I
Stages of meiosis I:
1. Prophase I: is divided into 5 stages, which are:
a. Leptotene during which the chromosomes become visible as long thin
threads showing bead-like structures called chromomeres.
b. Zygotene during which the chromosomes thicken and seen to be doubled.
The maternal and paternal homologous chromosomes pair along their entire
length side by side by a process called synapsis. This process is associated
with the formation of synaptonemal complex between the paired
chromosomes. Pairing of sex chromosomes occurs between their short arms
end to end.
c. Pachytene during which the chromosomes condense greatly and become
shorter. Homologous chromosomes appear as bivalents each of which is
composed of two visible chromosomes or four chromatids and hence called
tetrads. At this stage crossing over is believed to occur.
d. Diplotene the chromatids of each tetrad start to separate except at some
discrete points of contact called chiasmata which are the sites of crossing
over.
e. Diakinesis during which the chromosomes become more coiled and dense,
and the two chromatids joined by chiasmata start to separate from each other.
Nucleoli disperse and nuclear envelope breaks down.
2. Metaphase I:
The spindle fibers attach to chromosomes which arrange themselves at the
equator of the cell.
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3. Anaphase I:
One whole chromosome from each pair moves to one pole of the cell. No
centromeric division occurs in anaphase I.
4. Telophase I:
Nuclear envelope is re-assembled around the two separate sets of
homologous chromosomes. Cleavage of cytoplasm occurs giving two
daughter cells, with half chromosome number of the parent cell
chromosomes.
Meiotic Crossing Over:
During pairing of maternal and paternal homologous chromosomes,
recombination takes place by forming bridges or chiasmata.
At chiasmata, the two chromosomes break at identical points and the bropken
segments rejoin resulting in switching of distal segments from one
homologous chromosome to another. In this process, there is no loss of
genetic materials, but new gene combination will be produced leading to
genetic diversity.
MEIOSIS II
There is no DNA synthesis in its short interphase. It is similar to mitosis with
the difference that cells undergoing meiotic division as well as the resulting
daughter cells are having 23 chromosomes (the haploid number).
The Consequences of Meiosis:
1. Reduction of chromosome number.
2. Segregation of alleles
3. Genetic diversity due to gene exchange.
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Differences Between Meiosis and Mitosis:
a. Meiosis occurs in germ cells while mitosis occurs in somatic cells.
b. Meiosis has two successive divisions but mitosis is a single division.
c. Pairing of homologous chromosomes and crossing over occurs in meiosis
d. Meiosis reduces the chromosome number from diploid (46) to haploid
(23) number.
e. No centromeric division in meiosis I.

Cell Death
Two forms of cell death are known, necrosis and apoptosis. Necrosis is a
pathological accidental process that results from anoxia, mechanical injury or
exposure to toxins. Necrotic cells and their organelles swell and finally burst
releasing their content into the extracellular space. The debris of necrotic
cells is taken up by macrophages.

Apoptosis:
Apoptosis is a programmed cell death; it is not caused by an injury.
 Apoptosis is common in cells with short life span.
 Unlike necrosis, apoptotic cells do not swell, but they decrease in size
 The nuclei, show signs of pyknosis (become small and darkly-
stained).
 The chromatin is broken into short pieces by DNA endonucleases.

59
  Then nuclei disappear (karyolysis), and the cytoplasm breaks into
large vesicles or blebs which detach from the cell surface.
 Caspases digest relevant cellular materials and structures. [Caspases =
cysteinyl aspartate-specific proteinases]
 Finally the shrunken cell and its remnants are phagocytosed by
macrophages.
Importance of apoptosis:
 It is important to protect the cells and regulate their number and
balance cell proliferation in mature renewing tissues, such as blood
and epithelia.
 Apoptosis is important in morphogenesis and shaping of the embryo,
although apoptosis is present in the tissues of normal adults.
 Most cells are able to activate their gene program of apoptosis when
major changes of DNA occur just before the tumor cells start mitosis.
 Most T lymphocytes are destroyed inside thymus gland (by activation
of their apoptic DNA programs) before releasing into circulation to
avoid their serious damaging effect on other cells. Also, one strategy
of development is to overproduce cells, then select, e.g., for the
survival of lymphocytes which are reactive to non-self antigens.

60
 EPITHELIUM
Epithelium is the tissue that covers or lines the free surfaces of the body. It is
one of four fundamental body tissue with the other three basic tissues
(muscular, nervous, and connective tissue).

General features of epithelial tissue:

1. Epithelium is composed mainly of cells
2. Very small amounts of intercellular substance.
3. The cells have strong adhesion due to adhesion molecules, membrane
interdigitations and intercellular junctions.
4. The cells are usually arranged in one or more layers.
5. The epithelium has a free apical (pole) surface which is either exposed to
air or fluid, basal pole resting on basement membrane and lateral surfaces
facing the neighboring cells.
6. Its lower surface is resting on a basement membrane, which rests upon an
underlying connective tissue. In the digestive, respiratory and urinary
systems, the underlying connective tissue is called lamina propria which
supports, binds and nourishes the epithelium. At the site of contact between
epithelium and connective, the latter shows evaginations called connective
tissue papillae.
7. Epithelium is an avascular (not penetrated by blood vessels), it depends on
diffusion from the underlying blood supply of the connective tissue for
exchange of nutrients, gases and waste products.
8. The epithelial cells are penetrated by nerve fibers
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9. The epithelium has high regeneration ability
10. It develops from any of the three embryonic layers (endoderm,
mesoderm, and ectoderm)

Classification of epithelial tissue:

Epithelium is classified according to:
A. Number of layers:
1. Simple: made of one layer only
2. Stratified: made of more than one layer.
B. Function into:
1. Surface (covering or lining)
2. Glandular
3. Myoepithelium
4. Neuroepithelium

Simple Epithelium
This epithelium is made only of one layer of cells and all the cells touch the
basal lamina. It could be simple proper where all cells reach the free surface
or pseudostratified where the basal cells fail to reach the free surface.
Simple epithelium has mainly absorptive and secretory functions.
Types:
I. Simple squamous epithelium: it is a pavement epithelium that is made of
one layer of flattened scale-like cells, with flat nuclei and very small amount
of cytoplasm.
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Sites and functions of simple squamous:
i. Lining the lung alveoli to allow gas exchange, they are called
pneumocytes
ii. Lining the serous cavities to give smooth surface and prevent friction
during movement; it is called mesothelium
iii. Lining the blood vessels to provide smooth lining, where called
endothelium
iv. Form the parietal layer of Bowman`s capsule to facilitate diffusion and
filtration.

2. Simple cuboidal epithelium: It is made of one layer of cells, where the

height of each cell is equal to its width. The nuclei are rounded and centrally
located.
Sites of simple cuboidal:
 Thyroid follicles
 Pancreatic acini, exocrine part
 Secretory units of salivary glands
 Proximal and distal tubules of kidney

3. Simple columnar epithelium: It is made of one layer, where the height

of its cells exceeds their width, and their nuclei are oval and located in the
lower one-half of the cells. The simple columnar epithelium could be
partly ciliated (uterine tube and uterus), or non-ciliated (GIT). The non-

63
ciliated may be secretory (stomach), or absorptive with brush border as in
small intestine and gall bladder.

4. Pseudostratified columnar epithelium: the nuclei of this epithelium

appear in irregular layers giving the false impression of being stratified. In
this type of epithelium, all the cells rest on the basement membrane. Some
tall cells reach the free surface, and others, short basal cells do not reach the
free surface. This epithelium might be:
a. Ciliated present in upper respiratory tract plus goblet cells and
referred to as respiratory epithelium.
b. Non-ciliated lining the large ducts of salivary glands, ductus or
vas deferens and membranous part of the male urethra.

Stratified Epithelium
This type of epithelium has more than one layer of cells (layered) and its
surface cells are subject to wear; it protects against damaging mechanical and
chemical actions.
Stratified epithelium could be squamous keratinized (skin), non-keratinized
(mucous membranes), parakeratinized (tongue papillae), cuboidal, columnar
or transitional (urinary tract).
Stratified epithelium is named according to the shape of surface cells even
though the underlying cells may be of different shape.

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The basal layer is made of columnar cells which rest on the basement
membrane; they are mitotically active and migrate upwards to replace cells
lost from the surface, or cells destroyed by apoptosis.
Cells above the base become polyhedral and are held together by many
desmosomes to resist the abrasive forces on this protective epithelium.
The lower surface of the epithelium is undulated.

1. Stratified squamous non-keratinized (moist or wet):

The surface cells of this epithelium are flattened and nucleated while the
middle layer is made of polyhedral cells and are held together by many
desmosomes to resist the abrasive forces applied to the epithelium.
The basal layer is made of columnar cells which rest on the basement
membrane; they are mitotically active and migrate upwards to replace the
cells destroyed or lost from the surface.
The lower surface of the epithelium is undulated and indented by vascular
papillae of connective tissue, except in the cornea.

Sites and functions:

Its main function is protection. It is present in:
 Mouth cavity, pharynx, esophagus
 Vagina and vaginal surface of uterine cervix
 Anal canal
 Cornea

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2. Stratified squamous keratinized or cornified (dry):
It is similar in its basal and middle layers to stratified squamous
nonkeratinized, but its uppermost layer has granular cells concerned with
forming special, dead cells solidly packed together as a surface keratin layer
for greater protection.
A good example is thick skin where stratum corneum of epidermis is very
thick, its most superficial cells are dead with no nuclei and the cytoplasm is
replaced by scales of proteins, keratin proteins.
The basal layer is made of columnar cells which rest on the basement
membrane; they are mitotically active and migrate upwards to replace cells
lost from the surface, or cells destroyed by apoptosis.
Cells above the base become polyhedral and are held together by many
desmosomes to resist the abrasive forces.
The lower surface of the epithelium is undulated and indented by vascular
papillae of connective tissue.
Protection is the main function of this epithelium.

3. Stratified cuboidal and stratified columnar:

Stratified cuboidal is lining the ducts of sweat glands. The epithelium
consists of two layers of cuboidal cells; the deeper layer rests on a basement
membrane.
Stratified columnar is rare, present in the conjunctiva, cavernous urethra
and in large excretory ducts of some glands. The surface layer is made of

66
columnar cells, and the basal cells are cuboidal. One or more rows of
polygonal cells are present between the basal and the surface layers.

Transitional epithelium (also called urothelium):

This epithelium is specially lining the organs of urinary system; where these
organs undergo major changes during filling and emptying.
In contracted or empty state, the epithelium may be made of 5-8 layers. The
surface cells appear as large cuboidal bulging into the lumen (dome-like)
with rounded free surface, sometimes binucleated and contain fusiform
cytoplasmic vesicles. The basal cells are smaller than the surface cells and
interdigitate with the overlying cells; no connective tissue papillae indent the
epithelium.
When the bladder is full or dilated (distended), the epithelium appears to be
only 2-3 layers thick, and the surface cells are flattened.
The underside of the epithelium has no connective tissue papillae.

General functions of epithelial tissue:

 Protection  Lubrication
 Secretion  Sensation
 Excretion  Reproduction
 Absorption  Gas and nutrient exchange

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 Examples of epithelium

Tissue Epithelium

endothelium of blood vessels Simple squamous

esophagus Stratified squamous, non-keratinised

stomach Simple columnar, non-ciliated

small intestine Simple columnar, with microvilli

large intestine Simple columnar

rectum Simple columnar

gallbladder Simple columnar

thyroid follicles Simple cuboidal

endothelium of lymph vessel Simple squamous

skin Stratified squamous, keratinized

ducts of sweat glands Stratified cuboidal

mesothelium of body cavities Simple squamous

ovaries Simple cuboidal , germinal epithelium

Fallopian tubes Simple columnar, ciliated

uterus Simple columnar, partly ciliated

vagina Stratified squamous, non-keratinised

ductuli efferentes Pseudostratified columnar

epididymis Pseudostratified columnar, with stereocilia

vas deferens Pseudostratified columnar

ejaculatory duct Simple columnar

bulbourethral glands Simple columnar

trachea Pseudostratified columnar, ciliated

cornea Stratified squamous, non-keratinised

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Proximal convoluted tubule Simple high cuboidal or low columnar, with microvilli

Collecting duct (kidney) Simple cuboidal

ureter Transitional

urinary bladder Transitional

prostatic urethra Transitional

membranous urethra Pseudostratified columnar, non-ciliated

penile urethra Pseudostratified columnar, non-ciliated

external urethral orifice Stratified squamous

N.B. Lower part of anal canal and anal orifice are lined with stratified squamous non-keratinized.

Basal lamina and basement membrane:

 Basement membrane was describing the PAS positive layer seen by
LM, and separating between connective tissue and the basal layer of
epithelial tissue.
 Basement membrane is made of basal lamina (secretd by epithelial
cells), and lamina reticularis (secreted by connective tissue cells).

The basal lamina is a thin supporting layer lying between the epithelium and
the underlying connective tissue visible only with the electron microscope.
By EM, the basal lamina is formed of collagen type IV, laminin and heparin
sulphate; it appears to be composed of two layers, lamina lucida and lamina
densa.
Lamina densa is an electron dense layer 20-100 nm thick, composed of
delicate network of fine fibrils.

69
Lamina lucida or rara is an electron lucent layer lying on one or both sides
of lamina densa.
Basal laminae are present in epithelial tissue and at the points of contact
between some cells and connective tissue such as muscle cells, fat cells and
Schwann cells; theses cells secrete the components of basal lanina.
In some tissues like lung alveoli and renal glomeruli, the basal lamina is
thicker than usual because of the fusion of the two opposing epithelial cell
layers which have no intervening connective tissue in between.
Basal laminae are attached to underlying connective tissue by anchoring
fibrils of collagen type VII. Basal lamina is composed of:
 Collagen type IV
 Glycoproteins: laminin and entactin
 Proteoglycans: heparan sulfate proteoglycan (perlecan).

Functions of basal lamina:

 Structural support to the epithelium
 Provides a barier which regulates the exchange of macromolrcules
 Passive filtration in kidney and capillaries.
 Influence cell polarity
 Regulate cell proliferation and differentiation
 Influence cell metabolism and cell migration
 Organize proteins in adjacent cell membranes
 Contains information important for cell-to-cell interaction.

70
  Important for reinnervation of denervated nerve and to establish new
myoneural junctions in muscles.

Epithelial polarity:
Polarity of cells means that these cells have their functions preferentially
directed towards one end. The epithelial polarity appears in:
1. Apical free specialized surface and basal surface resting on basal
lamina
2. Supranuclear position of Golgi complex
3. Accumulation of secretory products in the apical compartment
4. Localization of receptors for chemical messengers in the basolateral
compartment
5. Presence of tight junctions to prevent intermingling of integral
membrane proteins of various regions.

Nutrition of epithelial tissue:

Since the epithelium is not penetrated by blood vessels; it receives its
nourishment by diffusion of metabolites through the basal lamina and
lamina reticularis. Presence of papillae enhances the process of diffusion.

Innervation of epithelium:
Epithelium has rich sensory nerve supply since it is penetrated by nerve
fibrils. Cornea is the best example of rich sensory nerve supply.

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Renewal and Regeneration:
The epithelial tissue has a high regeneration power; its cells are renewed by
mitosis of the basal layer. The rate of renewal is variable; it is fast in
intestinal epithelium and cornea and slow in others like pancreas. In stratified
and pseudostratified epithelia, replacement of the shed layers takes palce by
mitosis of the germinal basal alyer.

Metaplasia:
It is a reversible transformation of one epithelial tissue into another type
due to abnormal circumstances.
Metaplasia is noted in these conditions:

 In heavy smokers a change from pseudostratified columnar ciliated to

stratified squamous in the larynx and trachea.
 Esophageal stratified squamous epithelium changes to gastric or
intestinal simple columnar (Barrett's oesophagus), after repeated acid
reflux.
 In chronic vitamin "A" deficiency the epithelia of bronchi (cuboidal)
and urinary bladder (transitional) transform into stratified squamous
keratinized epithelium.
 From transitional to stratified squamous in bladders with a stone, or
infested with worms.

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Epithelioid means an epithelium-like appearance of non-epithelial cells such
as secretory cells of muscular origin in the juxta-glomerular apparatus of the
kidney.

Specialization of epithelial cells

 Basal
 Lateral
 Apical or free surface

Basal specialization of epithelial cells:

 Folding (ion transport cells)
 Hemidesmosomes

Lateral specialization:
 Cell bridges with true cytoplasmic continuity: seen rarely, e.g.,
between spermatids.
 Folding
 Junctions

Apical specialization:
 Microvilli
 Cilia
 Stereocilia

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Cell attachment:
The attachment between the neighboring cells and the underlying
connective tissue is achieved by intercellular junctions. These attachments
provide:
 Mechanical strength
 Selective transport by preventing the flow of materials through the
intercellular space.
 Formation of intercellular compartments
 Communication (gap junctions).

Intercellular Junctions
The epithelial cells maintain extensive lateral cohesion apposition. Their cell
membranes are separated by narrow intercellular spaces occupied by
proteoglycans and glycoproteins called cell adhesion molecules (CAMs).
Cell to cell adhesion is due to the binding action of the integral membrane
glycoproteins and due to the lateral specialization forming intercellular
junctions. Beside their adhesive power, intercellular junctions prevent the
flow of materials through the intercellular space (paracellular pathway).
Types:
1. Tight junction (zonula occludens):
It is the most apical, belt-like structure completely surrounding the cell and
lying just below the free surface. The intercellular space is completely
obliterated, the outer leaflets of the adjacent cell membranes fuse giving a
pentalaminar structure. In freeze-fracture preparation (cryofracture), the
74
replicas show number of ridges and grooves denoting the fusion sites of the
adjacent membranes. The less the number of fusion sites, the more
permeability of epilethelial tissue. Its main function is to close the
intercellular space and prevent small and large molecules from passing
through the epithelium via paracellular pathway.
2. Zonula adherens:
It is a belt-like junction encircling the cell lying just below the tight
junction, with very narrow intercellular space. The most obvious feature is
the presence of dense cytoplasmic plaques associated with this junction,
with numerous microfilaments, intermediate filaments and spectrin,
inserted into it. The microfilaments arise from the terminal web present in
the apex of cytoplasm, where cytoplasmic organelles are excluded.
Terminal web imparts rigidity to the apex of the cell.
Zonula occludens and zonula adherens are used to be called as terminal
bar in LM.
3. Macula adherens (desmosomes):
The desmosomes are not encircling the cells, they are discrete disc-like
plaques scattered over the lateral surface of the cells with an identical one
at the surface of the neighboring cells.
The cell membranes are straight and separated by 30 nm space (normally
20 nm), sometimes with a line of dense material in the intercellular space.
There are groups of keratin intermediate filaments inserted into the
cytoplasmic face of the attachment plaques (made of 12 proteins, mainly
cadherins) in a hairpin-like manner.
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Desmosomes are the only type of junctions present in stratified squamous
epithelium of the skin and the intermediate filaments are made of the strong
cytokeratin to provide strong adhesion of the cells and give stability to the
epithelium. Cadherin, an adhesive protein controlled by Ca ions, also
shares in the adhesive power of desmosomes.
At the base of the epithelium, at the contact between the epithelial cells and
the basal lamina, there are numerous hemi-desmosomes (half
desmosomes), which bind the cells firmly to the basal lamina. The
attachment plaques contain proteins mainly integrins which are
transmembrane proteins with receptors for laminin and collagen type IV.
4. Gap junctions or connexons:
The gap junction is made of pairs of units, called connexons which are
present along the lateral wall of almost all cells except skeletal muscles.
Each connexon is made up of 6 proteins called connexins, joined together
to form hydrophilic channels with a central pore of 1.5 nm, connecting the
neighboring cells and permit the exchange of molecules having MW of
1500 Da or less (ions, amino acids, cAMP). Gap junctions have an
important role in coordinating the activities of cells such as heart
coordinated beats.

5. Fascia adherens: at intercalated discs of cardiac muscle.

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 Specializations of the cell free surface
The epithelial tissue shows numerous specialization or modifications of the
free surface of its cells such as:
 Striated border (microvilli), and glycocalyx
 Stereocilia.
 Cilia and flagella
1. Microvilli:
In light microscope, the glycocalyx and microvilli are called brush (in
proximal convoluted tubules) or striated border (in small intestine and gall
bladder). In EM, the brush border is found to be formed of slender finger-
like projections 1 um long and 0.08 um wide; there are about 3000/cell
surface. Microvilli are extensions of the cytoplasm and covered by cell
membrane and glycocalyx which is a PAS positive coat rich in
glycoproteins contents. Each microvillus contains a bundle of 20 – 30 actin
microfilaments which are cross-linked to each other and to the surrounding
plasma membrane by the protein villin. The basal ends of microfilaments
intermingle with those of terminal web. The main function of microvilli is
to increase the surface area for absorption.

2. Stereocilia:
They are not true cilia, non-motile, long and branched microvilli present in
ducts of epididymis. They increase the surface area and facilitate the
movement of macromolecules between the cell and the surrounding
environment.

77
3. Cilia and Flagella:
Cilia are motile cell processes, arranged in rows in the apical cell surfaces,
beating in constant direction in the living state. Axoneme forms the internal
structure of both cilia and flagella, which are surrounded externally by the
plasma membrane. Cilia are shorter (5 - 10 um long and 0.8 um wide) than
flagella (50 - 100 um); they are inserted into basal bodies at the cell apex.
Flagellum is present in human body in the sperm, one per cell, while cilia are
present in large number (250 cilia) per cell.
In the living state, cilia have rapid coordinated back and forth movement to
allow a current fluid to be propelled in one direction over the ciliated
epithelium. The energy needs for this process is supplied by ATP.

Glandular Epithelium
This one type of epithelial tissue is specialized to produce secretion stored
in membrane-bound vesicles, secretory granules.
Glandular epithelium is classified into either endocrine or exocrine
according to the presence or absence of duct systems.
A. Endocrine glands
These are ductless (have no ducts), and their secretion (hormones) is
carried directly to their site of action by the bloodstream, and not by ducts.
The endocrine cells either form anastomosing cords with dilated blood
capillaries in between such as adrenal gland, or follicles filled with
secretory products like thyroid follicles, or irregular aggregates like
pituitary.
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B. Exocrine glands
These glands are composed of secretory units and duct systems. They might
be classified in different ways.
1. According to number of cells
a. Unicellular glands like goblet cells which are the best example, they
are present in large number in the lining epithelia of large intestine and
the respiratory tract.
b. Multicellular glands which form most of the glands of the body,
example salivary glands.

2. According to the mode of secretion

a. Merocrine glands (exocrine part of pancreas), where the secretory
products leave the cells by exocytosis with no loss of cellular
components.
b. Holocrine glands (sebaceous glands), in which the secretory product
is shed with the whole cell, a process which involves destruction of
the secretory cell.
c. Apocrine glands (mammary and axillary sweat glands), in which the
secretory product is discharged with parts of the apical cytoplasm.

3. According to the type of secretory product

a. Serous glands which are composed of serous acini and secrete watery
secretion e.g. parotid gland and serous acini of pancreas. The cells are

79
 usually pyramidal in shape or cuboidal, with round nuclei, basal
striations, apical secretory granules, and narrow lumen of the acini.
b. Mucous glands which are composed mainly of mucous acini and
secrete viscid mucus secretion (glycoprotein), e.g. sublingual salivary
gland. The cells are usually cuboidal to columnar in shape, pale
staining cytoplasm, flat basal nuclei and wide lumina of the mucous
acini.
c. Seromucous glands (mixed) which contain both serous and mucous
acini e.g. submandibular salivary glands.

4. According to the shape of the ducts which transport the secretion

a. Simple exocrine glands if the ducts are unbranched.
b. Compound exocrine glands if the ducts branch repeatedly.
5. According to the shape of secretory portion which contains the cells
responsible for secretion.
a. Tubular:
 Simple unbranched: intestinal glands
 Simple branched: fundic glands of stomach
 Compound: kidney
b. Coiled, simple: sweat glands
c. Acinar ( alveolar ):
 Simple branched: sebaceous glands
 Compound: mammary glands

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 d. Tubulo-acinar ( tubulo-alveolar ):
 Compound: salivary glands and exocrine pancreas

Note that: Adenocarcinomas are malignant tumors derived from glandular

epithelium.
Formation of glandular epithelium:
The cells of covering epithelia proliferate and penetrate the underlying
connective tissue.
 If a contact with the surface is maintained through a duct system,
exocrine glands are formed.
 If no contact is maintained with the surface and no ducts are formed,
endocrine glands develop; their cells are arranged as anastomosing
cords (adrenal) or as follicles (thyroid). The cords or follocles are in
rich blood supply and their secretion is carried to target tissue by the
blood stream.

Controle of glandular activity:

Glandular secretion is under neural and endocrine control mediated by
chemical messengers.
 The exocrine secretion of pancreas is stimulated by the
hormones secretin and cholecystokinin.
 Salivary gland secretion is under neural control.

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 Myoepithelium
This is a specialized type of epithelial cells which embrace the glandular acini
(mammary, sweat and salivary) and are located between the basal lamina and
the basal pole of secretory or ductal cells. They are connected to each other
and to epithelial cells by gap junctions and desmosomes. They are branched
or spindle-shaped cells containing cytokeratin intermediate filament, myosin
and actin filaments; their function is to contract around the secretory portion
and thus helping to propel the secretory products.

Neuroepithelium
It is specialized type of epithelium responsible for sensory perception of
smell (olfactory epithelium) and taste (taste buds).
The taste buds in the tongue papillae have three types of cells, supporting,
basal and sensory cells.

Examples of specialized epithelial cells

Once the cells differentiate, they start to get morphological and
physiological characteristics to optimize their functions. Some of these cells
will be studied in detail.
1. Ion transporting cells
Such cells are present in the intestine, proximal convoluted tubules of kidney,
the striated ducts of salivary glands and gall bladder; these cells transfer
sodium across the cells from apex to base (transcellular transport) using ATP

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as an energy source (active transport). In choroid plexus and ciliary body, the
flow of transport is from base to apex.
Characteristic features:
 The apices of these cells are freely permeable to sodium ions
which are accompanied by chloride ions and water.
 The basal cell membrane of these cells is highly folded
 Their lateral walls interdigitate with neighboring cell
walls.
 The basal infolding and lateral interdigitations are rich in
vertically oriented mitochondria and ATPase.
 Tight junctions are present to prevent the back diffusion.

2. Cells that transport by pinocytosis

Pinocytosis is a function of cell membrane that allows the transport of
macromolecules across the plasma membrane. Pinocytosis is well
developed in the lining endothelium of blood and lymphatic vessels and in
mesothelia lining the body cavities. The cells have few organelles and
abundant pinocytotic vesicles on the cell surface as well as in the
cytoplasm; such a phenomenon was confirmed by injection of ferritin or
colloidal gold the examined by EM for electron-dense particles in the
vesicles.

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3. The diffuse neuroendocrine system
These are chemical messenger producing cells; they are endocrine cells
present among non-endocrine ones. They produce messenger substances
which influence the activities of other cells; their cytoplasm contains
hormones or biogenic amine (epinephrine, norepinephrine or serotonin).
They used to be called APUD for amine precursor uptake and
decarboxylation. They are also called argentaffin and argyrophil cells due
to their affinity to silver stain. They are localized by immunohistochemical
and cytochemical techniques.
They are derived from the neural crest of embryonic nervous system and
widely spread (about 35 types) in respiratory, urinary, and gastrointestinal
system, thyroid and hypophysis.
These cells are classified into three groups:
a. Neurocrine cells such as neurons which secrete their chemical
messenger neurotransmittor at the synapse.
b. Paracrine cells, which secrete their message that diffuses through
the extracellular fluid and acts upon the neighboring cells without
passing through vascular system (mast cells).
c. Endocrine cells which secrete their messages into blood to reach
directly the target cells.

Examples of DNES cells:

 G cells of duodenum produce gastrin
 I cells of small intestine produce cholecyctokinin
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  S cells of small intestine produce secritin
 D cells of stomach, duodenum and pancreas produce somatostatin
 C cells of thyroid produce calcitonin
 B cells of pancreas produce insulin

Note that: Apudomas are the tumors derived from the cells of DNES.
4. Steroid secreting
These cells are found in testes, ovaries and adrenals; they are endocrine
cells that synthesize and secrete steroids. They have special features:
 They are rounded or polyhydral, with acidophilic cytoplasm
containing lipid droplets
 They have abundant SER and have the enzymes for
cholesterol synthesis and the enzymes to transform
mitochondrial pregnenolone into sex hormones.
 In these cells mitochondria tubular cristae (for energy) and have
the enzymes important for cholesterol metabolism by cleaving
its side chain.
 SER and mitochondria have the necessary enzymes to
synthesize cholesterol and steroid hormones.

5. Serous cells
The best example for these cells, are the pancreatic and parotid acinar cells.
They have characteristic features such as:

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  Polyhydral or pyramidal cells with round centrally located nuclei and
basophilic cytoplasm rich in RER and polysomes.
 They exhibit polarity, secretory granules in their apical compartments
and supranuclear Golgi complex.
 These granules empty their contents by exocytosis. The movement of
the granules is influenced by cytoskeleton, and their fusion with the
cell membrane is controlled by specific proteins.

6. Mucus –secreting cells:

Goblet cells of small intestine are the best examples. They have expanded
apical part and narrow basal part.
Goblet cells secrete mucus which lubricates and protects the surface of
epithelium. Their mucins, glycoproteins, differ according to the site of
secretion.
Glycoproteins (mucins) are produced by cells of the small and large intestine,
stomach, salivary glands, respiratory and genital tracts.
The apical part of the cell appears as a cup-shape, filled with secretory
granules containing mucins.
The base of the cell contains the nucleus, well developed Golgi complex just
above the nucleus and plenty of RER where monosaccharides are added to
proteins by glycosyl transferase enzyme. Mucins are hydrated after their
release to become viscous and lubricating mucus.

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 CONNECTIVE TISSUE
Connective tissue connects and binds organs thus supporting the body.
Unlike epithelium, connective tissue is made of fibers, ground substance,
and cells. Connective tissue cells and fibers are embedded in the
extracellular matrix. Classification of connective tissue is based on:
 Cellular and extracellular components
 The special function
Classification of connective tissue is different in different textbooks.
Connective tissue is classified into three categories:
I. Connective tissue proper including loose and dense (dense regular
and dense irregular).
II. Connective tissue with special features: adipose, elastic, reticular,
hematopoietic and mucous.
III. Supporting connective tissue: cartilage and bone.
In other textbooks, connective tissue is classified also into the following
three main types:
I. Connective tissue proper (loose and dense)
II. Embryonic connective tissue (mesenchymal and mucous)
III. Specialized connective tissue:
 Adipose CT
 Blood
 Bone
 Cartilage

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 Embryonic Connective Tissue
It is the type of connective tissue appearing in embryos and early fetal
development. It is composed of cells, fibers and jelly-like ground
substance.
In later development, the cells and extracellular matrix become specialized
for various tasks, and the matrix comprises amorphous ground substance
reinforced by specialized fibres.
Types:
1. Mesenchymal connective tissue:
It is a filling-in tissue, mesodermal in origin, composed of cells and matrix.
The mesenchymal cells are elongated, branched with cytoplasmic processes
and very little cytoplasm and large oval vesicular nuclei with prominent
nucleoli. The cells are actively dividing and show many mitotic figures.
The extracellular substance is abundant and viscous containing few fine ill-
defined collagen fibrils, and fetal blood vessels containing fetal blood cells.
It is the origin of all connective tissue, blood cells, endothelial cells and
muscle tissue.
2. Mucous connective tissue:
It is composed of stellate-shape fibroblasts, abundant ground substance
(mainly hyaluronic acid) and few fine collagen fibers. It is present in
umbilical cord and called Wharton`s jelly and also present in the pulps of
young teeth. The cord is covered externally with amnion with a simple
cuboidal or simple squamous epithelium; the cord has two umbilical
arteries and one umbilical vein.
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The umbilical blood vessels are lined with simple squamous epithelium.
Connective tissue proper together with bone and cartilage form the adult
connective tissue. It originates from mesoderm and has few cells and
fibers, and large amount of intercellular substance.

Functions of connective tissue:

 Structural, responsible to keep the structural interrelationship of the
various parts of the body.
 Protection by forming the capsules around body organs and
supporting their internal architecture
 Filling the spaces between the different organs
 Carries the blood vessels, nerves and lymphatics to organs
 Defensive mechanism against micro-organisms.
 Repair of wounds by fibrosis (formation of irregular collagenous
scar tissue).

Loose connective tissue

It is the most abundant type of connective tissue, and the best to study,
because it contains all types of connective tissue cells and fibers. It looks
loose, areolar and irregularly arranged.
It is found in:
 Subcutaneous region.

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  Between the muscle fibers and along nerves, blood and lymphatic
vessels and in glands and mucous membrane.
 Mesentery and serous linings.
 Forms the stroma of most organs.
Loose connective tissue is composed of:
 Ground substance
 Fibers
 Cells

The Ground Substance

The ground substance (extracellular or intercellular) is an amorphous,
highly hydrated, colorless, transparent and homogenous substance that fills
the spaces between cells and fibers.
It is a mixture of glycoproteins and proteoglycans; it is secreted by
fibroblasts and acts as a lubricant and as a barrier against penetration of
bacteria and other microorganisms.
The ground substance is composed mainly of:
 Glycosaminoglycans
 Proteoglycans
 Structural glycoproteins (multiadhesive)

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Glycosaminoglycans (GAGs): previousely were called acid muco-
polysaccharides due to their mucus-like nature. They are located on the
cell surface or in the extracellular matrix.
Glycosaminoglycans consist of linear polysaccharide molecules composed
of repeating pairs of disaccharides of uronic acid (glucuronic or iduronic
acid) and hexosamine (glucoseamine or galactoseamine).
They bind to a long protein core to give a proteoglycan molecule, with
many long side chains of disaccharides sticking out giving the appearance
of test tube brush.

Glycosaminoglycans include two groups:

 Non-sulfated (hyaluronic acid) present in umbilical cord, synovial
fluid and vitreous humor of the eye.
 Sulfated glycosaminoglycans act as polyanions and are strong
hydrophilic due to presence of sulfate, hydroxyl and carboxyl
groups. They are highly viscous and when hydrated, their
molecular size is highly increased and the molecules occupy
larger areas. Types of sulfated GAGs:
1. Dermatan sulfate present in skin, tendon and advetitia of the
aorta.
2. Keratan sulfate present in cornea, cartilage matrix and
intervertebral discs (nucleus pulposus and annulus fibrosus).
3. Chondroitin sulfate present in cartilage matrix, bone, skin, media
of aorta, cornea, umbilical cord, notochord.
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 4. Heparan sulfate present in aorta, lung, liver and basal lamina.
5. Heparin (also sulfated) present in granules of mast cell and
basophils.
Proteoglycans:
Proteoglycans are formed of fibrillar protein core associated with the four
sulfated glycosaminoglycans, dermatan sulfate, heparan sulfate, keratan
sulfate and chondroitin sulfate.
In cartilage, the molecules of proteoglycans bind the chains of hyaluronic
acid producing larger proteoglycan aggregates.
Proteoglycans differ from glycoproteins in: their fibrillar core proteins
(globular proteins in glycoproteins); lack of branching of their sugar chains
(branched in glycoproteins) which are usually their longer sugar chains, and
more acidic or negatively charged.
The protein part of proteoglycans is synthesized in RER, glycosylation
starts in RER and is completed in Golgi complex.
Types of proteoglycans:
 Aggrecan: present in extracellular matrix of cartilage
 Cell surface proteoglycans such as: Syndecan and fibroglycan
present in epithelial cells. The core protein of syndecan spans the
whole thickness of cell membrane.

Structural glycoproteins (multi-adhesive) are mainly made of globular

protein molecules bound covalently to the branched chains of mono-

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saccharides (carbohydrates), there are several types, fibronectin and laminin
are predominant:
 Fibronectin: secreted by fibroblasts and some epithelial cells; it
helps in deposition and orientation of collagen and mediates normal
cell adhesion and migration. It has binding sites for cells, collagen
and glycosaminoglycans.
 Laminin is a non-filamentous, large glycoprotein secreted by
epithelial and glial cells; it helps in adhesion of epithelial cells to
laminin-rich basal lamina.
 Fibrillin: secreted by fibroblasts and helps to enhance aggregation of
other components of extracellular matrix.
 Entactin secretd by epithelial and glial cells; it binds laminin to
collagen type IV.
 Tenascin is also secretd by epithelial and glial cells; it is present in
embryo.

Integrins are transmembrane proteins on cell membrane of connective tissue

cells acting as matrix receptors to bind to collagen, fibronectin and laminin
mediated by the intracellular proteins, paxillin, vinculin and talin.

Physical properties and functions of the ground substance

The ground substance contains a small amount of tissue fluid similar to

blood plasma in composition where it stores about one-third of body

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plasma proteins. Being viscous and negatively charged the ground
substance:

 attracts cations, and restricts the movement of water, ions and

molecules (and bacteria), thus controlling transport through the CT to
the cells and other tissues
 reforms the matrix, making it gel-like, or firmer in cartilage, where the
proteoglycans are bound along a hyaluronic acid backbone
 binds CT fibers to one another, influencing their strength and function.
 acts as a reservoir for growth factors and other agents controlling cell
behaviour.

Clinical notes:
 Degradation of proteoglycans is done by specific lysosomal
enzymes, difficiency of these enzymes leads to a group of lysosomal
storage disease
 The bacteria that produce hyaluronidase enzymes have great
invasive power to connective tissue since they reduce connective
tissue viscosity.
 Pathological increase of tissue fluid is called edema. Normally,
there a small amount of fluid in the extracellular matrix; the tissue
fluid is similar to plasa protein in composition and it stores about
one third the body plasma proteins.

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 Fibers of Connective Tissue
There are three types of fibers:
 Collagen
 Reticular
 Elastic.

Collagen Fibers
Collagen is the most abundant protein in the body and it is made of more
than 25 subtypes. They appear whitish or colorless and wavy in fresh
condition. Collagen forms about 30% of dry body weight; its main
function is to give the tissue rigidity, flexibility and strength.
Types and sites of collagen:
The are several types of collagen, type I, II, III, IV, V and VII are the most
important for medical histology course.
 Type I: is the most abundant, found in dermis, tendons, ligaments,
dentin, bones and fibrocartilage. It is secreted by fibroblasts,
odontoblast and osteoblasts; it is resistant to tension.
 Type II is found in hyaline and elastic cartilage and vitrous body,
dispersed in the ground substance; Secreted by chondroblasts. It is
resistant to pressure.
 Type III (reticulin), found in reticular tissue and coats of blood vessels
with smooth muscle, stains black with silver stains. It is secreted by
fibroblasts.

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  Type IV: non-fibrellar mesh-like structure present in basement
membrane secreted by endothelial, epithelial, muscle and Schwann
cells (not by connective tissue).
 Type V in fetal membranes and blood vessels, secreted by
fibroblasts.
 Type VII is found in anchoring collagen fibrils binding collagen
fibers to basal lamina; secreted by epithelial cells.
Based on its function, collagen is divided into 4 groups:
 Collagen forming long fibrils where its molecules aggregate to form
long fibrils, as seen in collagen type I, II, III, V, and XI.
 Collagen associated with fibrils which form short fibrils to bind
collagen fibrils together; these are collagen type IX, XII and XIV.
 Network forming collagen like collagen type IV which forms the
network of basal lamina.
 Collagen forming anchoring fibrils like collagen type VII which
anchors the collagen fibers to basal lamina.

Charcteristic features of collagen fibers:

 Collagen is found as thick wavy bundles 10 - 100 um in diameter.
The bundles branch but the fibers do not. Bundles are made of
fibers which, in turn are made of fibrils.
 The individual collagen fibers (1-20um) appear acidophilic in H&E
stain.

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  In EM, collagen fibers appear to be composed of packed fine fibrils
20 - 90 nm in diameter with cross striations at periodic distance of
64 nm.
 The dark striations on the fibrils are due to deposition of lead stain
on free chemical groups in tropocollagen subunits, which are
arranged in a stepwise manner.
 A tropocollagen molecule is composed of three interwined and
cross-linked helical polypeptide chains (two α α 2) rich in
glycine, prolyne and hydroxyproline.
 Collagen is digested by collagenase enzyme and yields gelatin on
boiling.
 Abnormalities in collagen formation leads to pathological conditions
such as: fibrosis and keloid formation due to excess collagen
formation, vitamin C deficiency leads to impaired collagen formation
and causes gum ulceration and hemorrhage, a condition known as
scurvy.

Collagen staining:
 H & E stains collagen pink and sometime orange; it is the routine
stain
 Collagen (type I) often is present in bulk, and is stained selectively
by: blue in Mallory's method, light green in Masson's, or red acid
fuchsin in van Gieson's.

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  Mallory's, Masson's and van Gieson's trichrome methods distinguish
collagen from muscle, and also react with the nuclei and cytoplasm
of other cells.
Collagen biosynthesis:
Its synthesis follows the same way of protein synthesis.
 Formation of mRNA for each of the three α-
 Polypeptide αchains of preprocollagen are synthesized and
assembled in RER of fibroblasts.
 Hydroxylation of proline and lysine takes place in RER by peptidyl
proline hydroxylase and peptidyl lysine hydroxylase enzymes.
 Hydroxylation step is followed by glycosylation of different amount
of galactose according to collagen type.
 The α chains when synthesized are having extra peptides called
registration peptides on both ends to help correct formation of the
triplet helix and prevent premature intracellular conversion of
procollagen into collagen fibrils.
 Procollagen molecules are transferred to Golgi complex by transfer
vesicles where procollagen is packaged into secretory vesicles and
released to extracellular space by the process of exocytosis where
the extra part of the molecule (registration peptide) is cleaved by
procollagen peptidase transforming procollagen into tropocollagen
molecules.

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  Tropocollagen aggregates and fibril formation takes place outside
the cell followed by cross-linking to attain the required strength.
 Hydroxyproline helps to stabilize the triple helix by forming
hydrogen bonds between the polypeptide chains which is re-inforced
by covalent cross-linking between tropocollagen molecules.

Reticular Fibers
They are very thin fibers with a diameter of 0.5 - 2 um; they are composed
of collagen type III plus glycoproteins and proteoglycans. They stain black
with silver (argyrophilic), and PAS positive due to their high contents of
glycoproteins.
By EM: they appear like fine collagen fibres, having the same 64 nm-
repeating cross banding and made of fine fibrils, loosely packed and bound
together by interfibrillar bridges of proteoglycans and glycoproteins
creating flexible networks of expansible organs such as blood vessels,
spleen, liver, uterus and intestines to maintain theirstructures.
Function:
 Reticular fibers form delicate network to support the cellular
components of endocrine glands, spleen, lymph nodes, bone
marrow, kidney and liver.
 Reticular fibres are abundant in smooth muscles and endoneurium.

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 Elastic Fibers
In fresh state, they appear yellow, wavy and thinner than collagen.
Elastic fibers are made of a core of elastin, elaunin and oxytalan fibers.
Elastic fibers, like collagen are rich in glycine and proline.
Differentiation of elastic fibers passes through 3 stages:
I. Formation of oxytalan fibers which are not elastic since they do not
contain elastin, but they are very resistant to pulling forces.
Oxytalan fibers are made of bundles of 10 nm microfibrils of
glycoproteins such as fibromodulin I, fibromodulin II and fibrillin.
II. Irregular deposition of elastin protein between oxytalan fibers
resulting in the formation of elaunin fibers.
III. Further accumulation of elastin protein until it fills the center of the
fiber bundle surrounded by a sheath of microfibrils. The resulting
fibers stretch easily in response to tension. Also, the presence of the
amino acids desmosines and isodesmosines when binds to lysine
adds to elasticity of the elastic fibers.
 Elastic fibers branch and anastomose; they can be digested by
pancreatic elastase enzyme and resist boiling, acid and alkali
extraction and digestion by the usual proteases.
 Elastic fibers stain pale pink with H&E, but stain specifically with
orcein and Verhoeff`s stains.
 Elastic fibers are produced by CT fibroblasts and smooth muscle
cells of blood vessels as a globular molecule, proelastin.

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Sites:
Elastic fibers are abundant in elastic tissue such as ligamentum nuchae,
alveolar septa of the lung and as fenestrated membranes in the media of the
aorta.
Function:
Elastic fibers impart the tissue the power to stretch and quick elastic recoil.

Cells of Connective Tissue

The cells of connective tissue are classified into two populations:
 Fixed cells which are relatively stable inhabitant of long-lived cells
including fibroblasts, adipocytes, fixed macrophages, pericytes,
endothelial and reticular cells.
 Free cells which are short-lived changing populations such as:
eosinophils, neutrophils, lymphocytes, free macrophages, pigment
cells, plasma cells and mast cells.
Mast cells, macrophages, plasma cells and leukocytes originate from
hemopoietic stem cells in bone marrow.

1. Fibroblasts are the most abundant cells in connective tissue; they

originate locally in connective tissue from undifferentiated mesenchymal
cells and spend their life there. Fibroblasts occur in young active and adult
less active forms (quiescent). The young active fibroblasts are large,
branched and irregular in shape, with abundant basophilic cytoplasm, well
developed RER and Golgi complex. Their nuclei are large, pale ovoid with
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prominent nucleoli. The mature, quiscent cells, fibrocytes are spindle-
shaped, smaller than the young, with little acidophilic cytoplasm and few
RER, and smaller darker nuclei. Fibrocytes revert to fibroblast state in case
of injury.
Function:
 Fibroblasts are responsible for synthesis of fibers (collagen, reticular
and elastic), glucoseaminoglycans, proteoglycans and multiadhesive
glycoproteins.
 Fibroblasts are responsible for wound healing and scar formation.
 In some circumstances, fibroblasts attain the features of fibroblasts
and smooth muscle cells, (myofibroblasts), and become contractile
due to increased amount of actin and myosin in their cytoplasm, thus
causing contracture of scar tissue in wound healing.
 Production of growth factor which influences cell growth and
differentiation.
Scleroderma is an inherited disease where excess connective tissue is
formed impairing blood flow and swallowing.

2. Macrophages are long-lived (few months), large (10-30 um), highly

phagocytic, with oval or kidney-shaped nucleus, acidophilc cytoplasm
containing visible vacuoles when examined by LM. In EM, the surface of
macrophages appears as irregular with protrusions and indentations due to

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their phagocytic activity. They have well developed Golgi, many
lysosomes, prominent RER, microtubules and microfilaments.
Macrophages develop from precursor cells in bone marrow which produce
monocytes, then migrate to connective tissue, mature and change to
macrophages.
When leukocytes cross the walls of venules or capillaries to invade an area
of inflammation, this process is called diapedesis.
Macrophages are responsible for removal of foreign bodies, bacteria and
aged broken cells (cell debris); they coordinate the inflammatory response
and healing by means of signalling peptides and proteins - cytokines.
In pathological conditions, macrophages may fuse to become multinuclear
giant cells with many nuclei, on facing a large object for digestion.
Phagocytic cells can be revealed and activated by vital stain injection (into
the living animal) of colloidal or particulate coloured matter, e.g., Trypan
blue or India ink. The phagocytic cells engulf and accumulate the stain in
their cytoplasm. They are also identified by their cell-surface glycoprotein
profiles. When activated by infection or foreign substance injection,
macrophages increase their size, their lysosomal conents and pahgocytic
activity.
Macrophages are antigen presenting cells, meaning that they have the
capacity to process antigen by digesting it into small peptides or small
molecular domains (epitopes) and present it on their surface to be recognized
by T-lymphocytes.

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Macrophages are distributed throughout the body forming the
mononuclear phagocyte system (reticuloendothelial system). The cells of
this system are called different names in different sites e.g:
 Macrophages of connective tissue and multinuclear giant cells
of connective tissue.
 Macrophages of lymph node, spleen and bone marrow
(dendritic cells).
 Alveolar macrophages (dust cells) in lungs
 Kupffer cells in liver
 Mesangial cells in kidney
 Microglia in CNS
 Osteoclasts in bone
 Langerhans cells in the skin
 Circulating monocytes
The cells of mononuclear phagocyte system are characterized by large size,
irregular outline, acidophilic cytoplasm, darkly stained eccentric oval or
kidney-shape nucleus, phagocytic vacuoles and many lysosomes.
Functions of macrophages:
 Ingestion of dead or aged cells and any other foreign particles, then
digestion by lysosomes hydrolytic enzymes, a process called
phagocytosis.
 In infection, macrophages change their morphology and metabolism
and secrete cytokines (interleukins) to combat bacterial invasion.

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  Participate in cell-mediated resistance to infection by bacteria,
viruses, fungi or metazoa.
 Participate in cell-mediated resistance to tumors.
 Participate in extra-hepatic bile production, iron and fat metabolism
and destruction of old erythrocytes.
 Produce chemotactic factors
 Antigen processing and presentation.
Cytokines:
These are molecules involved in intercellular communication by diffusion
through the blood to target cells that have specific receptors for those
molecules on their surface. Since lymphocytes secrete these substances,
they were called lymphokines, and because other cells secrete some of
these molecules, they were called cytokines.

3. Mast cells:
These cells are in connective tissue especially in skin dermis, digestive and
respiratory systems; they are large, with small spherical central nucleus
which is not seen well because of large number of dense basophilic
granules present in cytoplasm. RER and Golgi are well developed.
Mast cells originate from precursor cells in bone marrow; enter the
circulation, then cross the walls of capillaries and venules to enter the
connective tissue for proliferation and differentiation. They resemble blood
basophils, but originate of different stem cells; their granules give

105
metachromatic staining reaction with thionine or toluidine blue stains, since
they stain different from the stain applied and give a reddish-purple color
because they contain acidic sulfated glycoseaminoglycans (heparin).
The secretory granules have heterogenous interior and contain histamine,
(peripheral vasodilator and increase capillary permeability), heparin
(anticoagulant), neutral proteases, eosinophilic chemotactic factor of
anaphylaxis ECF-A, and slow reacting substance of anaphylaxis SRS-A or
leukotrines. Leukotrines are secreted from membrane phospholipids,
released into the extracellular space and not stored in cells; their secretion
act locally through paracrine mode.
Mast cells have surface receptors for immunoglobulin E (IgE) produced by
plasma cells, where most of IgE receptors bind the cell surface of basophils
and mast cells, leaving the rest of immunoglobulins in the plasma.
There are two subtypes of mast cells with no morphological differences:
 Connective tissue mast cells, 10-12 um in diameter; they contain
heparin. They are present in the skin and peritoneal cavity and
contain anticoagulant heparin.
 Mucosal mast cells, smaller than the previous cell type, present in
connective tissue of intestinal mucosa and lungs, their granules
contain chondroitin sulphate instead of heparin.
Functions:
 Secrete anticoagulant heparin

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  Secrete histamine which causes smooth muscle contraction, dilates
and increase permeability of blood capillaries.
 Secret eosinophilic chemotactic factor of anaphylaxis (ECFA), if
released in a previously sensitized person to the same antigen, leads
to a fatal anaphylactic shock.
 Starts the inflammatory response to noxious intruders.
4. Plasma cells are large ovoid with deep basophilic cytoplasm which is
very rich in RER. In routine histology preparations, a pale juxtanuclear
region containing Golgi complex and centrioles is seen. The nucleus is
spherical and eccentric, with a cart-wheel or clock-face appearance due to
regular arrangement of heterochromatin alternating with euchromatin.
Plasma cells are derived from B lymphocytes and live for 10 – 20 days.
Functions:
Plasma cells are responsible for production of antibodies in response to
invading antigens. Each antibody is specific for the antigen which
stimulated the production of that antibody.
5. Fat cells (adipocytes)
Adipose cells are large and spheroid with a diameter of up to 200 um,
distended by a huge fat droplet (glycerides of fatty acids). The cells look
empty in routine preparations due to dissolving of their fat contents during
processing with alcohols and fat solvents, but with osmium tetroxide
fixation it remains as black material. Some dyes will colour it, if it is
preserved by frozen sectioning. The nucleus looks flat and peripheral with

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a rim of cytoplasm giving the signet ring appearance. Fat cells are present
as single cells or in groups arranged into lobules forming adipose tissue.
They function as:
a. Storage of neutral fat which participates in carbohydrate and fat
metabolism.
b. Fat cells secrete specific enzymes for fat metabolism, and leptin, which
helps control energy balance, heat production and body fat mass.
c. Forming cushions for many organs.
6. Leukocytes
These white blood cells usually migrate from blood vessels (capillaries and
postcapillary venules) to connective tissue by a process called diapedesis.
In inflammation, diapedesis is greatly enhanced to defend the tissue against
bacterial invasion or chemical irritation. Leukocytes, the wandering cells
of connective tissue, except for the lymphocytes, do not return to the blood
circulation. In inflammation, the tissue is red, hot, swollen, pain and
disrupted functions; chemical mediators are released, and accompanied by
increase blood vessels permeability, cemotaxis and phagocytosis.
7. Undifferentiated mesenchymal cells
These cells are multipotential embryonic mesenchymal cells which can
give rise to other types of cells such as: fibroblasts, chondroblasts,
osteoblasts, endothelial and mesothelial cells.
8. Pigment cells or connective tissue melanocytes
Melanocytes originate from ectoderm of neural crest; they contain melanin
pigemts in their cytoplasm. They are found in skin epidermis, substantia
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nigra of brain and choroid of the eye. Their function is formation of
melanin.
9. Pericytes
These are branched cells with oval nuclei associated with blood capillaries
lying between the endothelial cells and their basement membrane. They are
considered as stem cells differentiating into fibroblasts and smooth muscle
cells.
10. Endothelial cells: These cells are simple squamous epithelium lining
blood vessel. They secret their own basal lamina and collagen type IV.
11. Reticular cells
These are branched cells with long cytoplasmic processes forming cellular
network. They are present with reticular fibers in the stroma of bone
marrow, lymphatic organs, kidney and liver. By histochemical techniques,
3 subtypes can be distinguished:
 Fibro-elastic type which produces the supporting fibers of lymphoid
tissue and bone marrow.
 Interdigitating type present in the T-zone
 Dendritic present in the B-zone and it is antigen-presenting.
Function: primarily supporting; could be phagocytic when stimulated.

DENSE CONNECTIVE TISSUE

This type of connective tissue is characterized by large number of fibers
and small amount of ground substance and few cells. According to the type
of fibers, it is classified into:

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  White fibrous connective tissue: regular and irregular
 Yellow elastic connective tissue

Regular white fibrous connective tissue

It is whitish in color, found in tendons, aponeuroses, ligaments, joint
capsules and fibrous coat of the eye.
It is made of parallel regularly arranged collagen fibers Type I aggregate in
primary bundles, with tendon cells arranged in between. The tendon cells
are fibrocytes with thin small flat nuclei and minimal cytoplasm. The
primary bundles further aggregate into larger bundles called secondary
bundles which are surrounded by a sheath of dense connective tissue which
contains blood vessels and nerves. The sheath is made up of two layers and
lined with simple squamous epithelium and contains a potential space with
a lubricant viscous fluid to allow free sliding movement of the tendon.
Function: supporting and withstanding mechanical stress.

Irregular white fibrous connective tissue

This connective tissue is made of white collagen fibers irregularly arranged
without definite orientation forming compact meshwork. This type of
tissue has very little ground substance and few cells mainly fibroblasts and
fibrocytes.
It is found in deep fascia, skin dermis, dura matter, fibrous capsule of
organs, tunica albuginea of testis, submucosa of GIT, sheath of large nerves
perichondrium and periosteum.
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Function: this tissue offers resistance to stress from all directions.

Dense yellow elastic connective tissue

It is composed of elastic fibers which, run in thick parallel bundles with
fibrocytes in between. Each elastic bundle is surrounded by small amount
of loose connective tissue of fine collagen fibers and fibroblasts.
Elastic tissue presents in ligamentum nuchae, fenestrated membranes of
large elastic arteries and suspensory ligament of penis.
Function: this type of tissue has the ability to stretch and elastic recoil.

Reticular connective tissue

It is a specialized type of loose connective tissue providing a framework for
lymphoid and myeloid organs and endocrine glands. It consists of reticular
fibers secreted by reticular cells which are modified fibroblasts. They are
large, stellate cells with pale staining nuclei. Reticular connective tissue
also contains phagocytic cells which monitor the materials passing through
sinuses and removes micro-organisms.

Adipose tissue
Adipose tissue is one variety of connective tissue with special feature being
made up mostly of fat cells or adipocytes. 15 – 20% in males and 20 – 25%
in females is made up of adipose tissue. Male and female fat distribution
depends on the sex hormones and adrenocortical hormones. There are two
types of adipose tissue, unilocular and multilocular
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I. Yellow (white) unilocular:
Its color varies from white to yellow depending on the diet and the
presence of carotenoids dissolved in fat droplets. This type of adipose
tissue comprises almost all fatty tissue in adult body. In human
newborn babies, white adipose tissue is distributed uniformly allover
the body, then changes distribution by growing.
Sites: Adipose tissue is present allover the body except eyelids,
scrotum, penis and auricle of external ear except the lobules; it is
found in subcutaneous tissue, around the kidneys, in mesentery, in
mediastinum, in axilla and inguinal region.

Adipose tissue is formed of large collection of fat cells which are

arranged in lobules separated by incomplete connective tissue septa
rich in nerve and capillary supply.
 The individual fat cell (adipocyte) appears spherical (50 -150 um in
diameter) when isolated but polyhydral in adipose tissue. Each cell
appears empty with a rim of cytoplasm, because the fat has been
removed by alcohol and xylene during preparation. Each
adipocyte is enclosed by a basal lamina.
 The nuclei are flat and peripheral or eccentric giving the signet ring
appearance. By EM, the rim of cytoplasm contains Golgi complex,
mitochondria, poorly developed RER, few ribosomes, SER and
pinocytotic vesicles; fat droplets are not enclosed by a membrane
but show many vimentin filaments at their periphery.
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Development (histogenesis) of white adipose tissue:
 Undifferentiated mesenchymal cells develop into lipoblasts.
 Lipoblasts which are branched and resemble fibroblasts are able to
accumulate isolated fat droplets in their cytoplasm.
 Isolated fat droplets fuse to form large single fat droplet resulting
in white fat cells.
Function:
1. Adipose tissue functions as shock absorber in palms and soles.
2. Conserve body heat by insulation and energy storage
3. Packs the orbital cavity and angles of joints.
4. Helps to shape the surface of the body. Fat deposition in the hips,
buttocks, and breasts are especially under the control of female sex
hormones.
5. Acts as a secretory organ such as lipoprotein lipase and leptin.
Leptin is a protein produced by fat cells whose receptors are present
in brain and other tissues; it controls the amount of adipose tissue in
the body by acting on hypothalamus to decrease food intake and
increase energy consumption.
II. Brown fat or multilocular fat:
It is called brown due to its appearance, because it has a large number
of blood capillaries and mitochondria containing large number of
cytochromes.

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The cells are polyhydral with central rounded nuclei containing many
separate lipid droplets and mitochondria with long tubular cristae. The
cells of brown fat are smaller and contain relatively more cytoplasm than
the white fat cells. The cells receive direct sympathetic innervations.
Sites: It is abundant in rodents and hibernating animals.
In humans, it is also present in the new born in the back of neck and around
the kidneys to produce heat, and is greatly reduced in adults. There is no
formation of multilocular adipose tissue after birth. No change of one type
of adipose tissue into anothe.
Development of brown fat:
 Undeffirentiated mesenchymal cells differentiate into lipoblast
which resemble epithelial cells.
 Lipoblasts accumulate several fat droplets in their cytoplasm
resulting in multilocular fat cells.
Function: mainly heat production (thermogenic). The inner mitochondrial
membrane of brown fat cells has a transmembrane integral protein called
thermogenin which allows the backflow of protons generating energy
dissipated as heat.

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 BLOOD
The blood is composed of fluid intercellular substance, the plasma (55%),
and formed elements suspended in the plasma which are red blood cells
(erythrocytes), white blood cells (leukocytes) and blood platelets
(thrombocytes).
Blood circulates in blood vessels performing these general functions:
 Transports gases, nutrients, cells and hormones throughout the
different parts of the body.
 Removes metabolic waste products.
 Controls body temperature.
 Controls acid-base balance.
 Defends body against microbial invasion

Blood plasma is an aqueous solution containing inorganic salts and plasma

proteins, which exert a colloidal osmotic pressure within the circulatory
system and controls the exchange of aqueous solution between plasma and
extracellular fluid.
There are three main types of plasma proteins:
 albumin,
 globulin
 lipoprotein
 fibrinogen
 prothrombin

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Albumin is the main components of plasma proteins; it acts as a transport
protein for fatty acids and maintaining the osmotic pressure of blood.
The globulins are α-, β-, and γ-globulins; gamma globulins are antibodies
and called immunoglobulins.
Fibrinogen is important for fibrin formation in coagulation; prothrombin is
important for blood coagulation.

The formed elements are best studied by blood smear to identify the
different kinds of blood cells.

Preparation of blood film (blood smear):

1. Clean your finger tip with alcohol and allow it to dry.
2. Prick your finger with a sterile lancet, a drop of blood will form.
3. Put a drop of blood in the middle of the right end of a pre-cleaned slide
near its edge. Put the slide on the bench. Always hold the slide from the
edges to avoid greasing the surface.
4. Use another slide with smooth edges as a spreader, and put its smooth
edge just in front of the blood drop at an angle of 45.
5. Move the spreader slide backwards so that it touches the drop of blood
which will spread across the line of contact between the two slides.
6. Push the spreader slide steadily (without stopping) and rapidly to obtain
a thin blood film. If the blood drop was the right size, the film will end in a
nice tail. The beginning of the film is called its head, and it is always too
thick.
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7. Immediately pick up the slide and wave about rapidly until it is dry.
With a pencil, write your name and date across the head of the blood film.
8. Put the slide containing the film across the bars of the staining rack.
9. Cover the film with 10 - 15 drops of Leishman's stain, putting the drops
in different places until the whole film is covered. This stain is formed of
eosin and methylene blue dissolved in methyl alcohol which acts as a
fixative.
10. After two minutes, add twice as many drops of the buffer as the stain
(20 - 30 drops). Mix well by blowing on with a pipette, leave it for 10
minutes. A fine metallic scum will form on the surface.
11. Drain the stain and buffer and wash with distilled water. Leave it until
completely dry.
12. Examine the blood film with an oil immersion lens (100X)

Formed Elements of Blood

A. Erythrocytes (Red Blood Cells or RBCs):
Erythrocytes are biconcave (for larger surface area) discs 7.5 um in
diameter, and 2.6 um in the greatest thickness at the rim and 0.8 um thick in
the center.

An adult male has an average of 5.5 million RBCs per cubic millimeter of
blood, while an adult female has 4.5 million RBCs per cubic millimeter.
Young erythrocytes, recently released by bone marrow into the circulation,
have rRNA, are called reticulocytes.
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Normally, reticulocytes constitute about 1% of the circulating erythrocytes.
Reticulocytes increase in number in hemorrhage and high altitudes.
The volume of packed erythrocytes per unit volume of blood; is known as
hematocrit, which is 40-50% in adult males, and 35-45% in adult females.
Human erythrocytes survive 120 days in circulation. The damaged RBCs
are removed by macrophages of bone marrow and spleen.
Crenation of erythrocytes may be seen where erythrocytes are shrunken and
have indented appearance when they are put in hypertonic solution.
On the other hand, when erythrocytes are put in hypotonic solution, they
swell and liberate hemoglobin into the medium, a process called hemolysis.
Sometimes RBCs adhere together forming rouleaux appearance in the thick
areas of the smear due to sticking of RBCs. This process occurs in stagnant
blood and does not occur in the normal circulating blood.
Erythrocytes are very flexible to allow adaptation to small irregular
capillary diameters.

When RBCs appear larger than 9 um in diameters, they are called

macrocytes.
When they appear smaller, less than 6 um in diameter, they are called
microcytes.
These abnormal cell sizes might occur in certain types of the anemias.

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Ultrastructure of RBCs:
The plasmalemma of RBCs is composed of 40% lipids, 50% proteins, 10%
carbohydrates. The phospholipid contents of each half of the lipid bilayer
are different; phophatidylcholine and sphingomyelin are more concentrated
on the outer half, while phosphatidylserine and phosphatidylethanolamine
are more concentrated on the inner half of the membrane.
Nearly half of the membrane proteins are transmembrane integral proteins
and the rest are peripheral proteins associated with the inner surface of
erythrocytes. The peripheral proteins form membrane skeleton (actin and
spectrin), thus determine the shape of erythrocytes and permit flexibility
when passing through capillaries.
Functions:
 RBCs contain hemoglobin (HbA in normal adult) that carries
oxygen in the form of oxyhemoglobin, and carbon dioxide in the
form of carbaminohemoglobin in a reversible manner in order to
liberate oxygen freely to the tissues. When oxygen combines with
carbon monoxide, an irreversible carboxyhemoglobin is produced;
intoxication and reduction of hemoglobin capacity to carry oxygen
occur.
 RBCs also contain the enzyme carbonic anhydrase, enzymes of
glycolysis and enzymes for hexose-monophosphate shunt pathway.

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Clinical notes:
 Anemia is a pathologic condition where hemoglobin concentration is
less than normal due to, hemorrhage, hemolysis, insufficient
formation and vitamin B12 deficiency.
 Sickle cell disease is an inherited alteration of hemoglobin, HbS,
where erythrocytes are inflexible, fragile with sickle shape and short
life span leading to anemia.

Blood group antigens:

The cell membranes of all cells have carbohydrate chains in the form of
glycolipids and glycoproteins. The human erythrocyte membranes have
two antigenic carbohydrate chains known as:
 Antigen-A
 Antigen-B
These antigens form the basis of ABO blood-group typing system. Some
people have antigen-A on their erythrocyte membranes, some others have
antigen-B, some have both and others have neither. Therefore, there are
four major blood groups: A, B, AB, and O.

All people have, in their blood plasma, antibodies against the antigens
which they do not have on their erythrocyte membranes.
Before blood transfusion, it is necessary to test the blood group, otherwise
intravascular agglutination and lysis of RBCs will occur.

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B. Leukocytes (White Blood Cells or WBCs):
These white blood cells are divided into two major groups:
 granulocytes (granular leukocytes)
 agranulocytes (non-granular leukocytes)
Normally, an adult male has an average of 6 - 10 thousands WBCs per
cubic millimeter of blood.
An increase of leukocyte number above the normal range is called
leukocytosis (more than 10,000 WBCs /cu.mm.), while a decrease below
5,000 WBCs / cu.mm. is called leukopenia.
In circulation, leukocytes are spherical and nonmotile but they become flat
and motile on encountering foreign bodies. They leave the circulation and
migrate to connective tissue by passing through the endothelial cells of
capillaries and venules by a process called diapedesis. The rate of
diapedesis increases in cases of infection and inflammation where the
inflamed tissue and microorganisms release chemical mediators which
attract specific cells by a process called chemotaxis.
Leukocytes have important role in cellular and humoral immunity against
foreign material and micro-organisms.

I. Granular leukocytes:
These are leukocytes which contain specific cytoplasmic granules and
lobed nuclei (2 – 5 lobes). The nuclear lobes although appearing separated,
they are connected to each other by thin delicate chromatin strands.
Granulocytes contain 2 types of granules:
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  Specific granules whose contents bind the neutral, the basic or the
acidic part of the stain, and according to which the granulocytes
were classified into neutrophils, basophils and eosinophils.
 Azurophilic granules are nonspecific and contain lysosomal
enzymes.
Granulocytes are nondividing cells with life span of few days, die in
connective tissue by apoptosis. They have poorly developed Golgi
complex and RER, and few mitochondria and can function in poorly
oxygenated areas.

1. Neutrophils
In healthy persons, these are the most numerous making up to 60 - 70% of
the total white blood cells. Polymorphs, is the term used by most
hematologists for neutrophils.
Neutrophils are short lived cells, with half-life in blood 6-7 hours and
1-4 days in connective tissue.
They are 12 - 15 um in diameter in blood smears and contain 3 - 5 nuclear
lobes.
In females, the inactive X chromosome appears attached to one side of
nuclear lobes.
The cytoplasm contains glycogen, to be broken into glucose to yield
energy, and two types of granules:

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  Specific neutrophilic granules, small in size, more abundant,
contain alkaline phosphatase, collagenase, lactoferrin, lysozyme and
antibacterial proteins phagocytins.
 Azurophilic granules which are primary lysosomes containing
hydrolytic enzymes
** Tertiarty granules (mentioned by some authors) containing
gelatinase enzymes which enhance phagocytosis.

The neutrophils with nonsegmented nuclei (band form or stabs) where the
nucleus is simply elongated or horse-shoe shaped are younger than
neutrophils having 3 - 4 lobe nuclei.
Barr body, a drumstick, inactive X chromosome is visible in 3% peripheral
blood of females.
In some pathologic conditions, neutrophils appear hypersegmented showing
nuclei with more than 5 lobes.
The number of neutrophils increases (neutrophilia) in acute bacterial
infection; they die after engulfing bacteria.
An increase in the peripheral blood of immature cells (bands) of the
granulocytic series is termed "shift to the left", while an increase of
hypersegmented is called a "shift to the right".
Function:

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  Constitute the first line of defense against microorganisms; they are
active phagocytes against small particles (microbes) hence, called
microphages.
 Clean up the inflamed and necrotic tissue by phagocytosis.
Once bacteria come in contact to neutrophils, in blood or tissue, they adhere
to the cell surface then surrounded and engulfed by pseudopodia. Part of
the cell membrane surrounds bacteria to give phagosome, where specific
granules fuse with it immediately and discharge their enzymes inside the
phagosome to kill and digest bacteria.

2. Eosinophils
Eosinophils constitute 2 - 4% of the total leukocytic count; they show
diurnal variation obtaining their highest number in the morning and the
least number in the afternoon.
They are 12 - 18 um in diameter, the same size or slightly larger than
neutrophils.
Eosinophils have bi-lobed nuclei hidden by the large, coarse, elongated and
refractile eosinophilic granules, and abundant glycogen particles. These
granules function as large lysosomes; they give positive reaction to
peroxidase and other lysosomal hydrolytic enzymes.
The granules have a dark lengthwise central stripe, crystalline core called
internum, containing the argentine-rich major basic protein, which
functions in killing the parasitic worms and gives the granules its bright
acidophilic color. The externum (matrix) is less dense material surrounding
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the internum. They are attracted by the substances released by basophils
and mast cells such as histamine and eosinophil chemotactic factor of
anaphylaxis. Eosinophils have receptors for IgE and produce eosinophil-
derived-inhibitor which inhibits mast cell degranulation and SRS-A (slow
releasing substances of anaphylaxis).
Corticosteroid hormones (of adrenal cortex) cause rapid drop in the number
of circulating eosinphils.
Function:
Eosinophils are important in engulfing antigen-antibody complexes, and
their number increases (eosinophila) following certain allergic reactions
and parasitic infestation.

3. Basophils
Basophils are smaller than the other granulocytes being 12 - 15 um in
diameter, and fewer in number making up to 0.5 - 1% of the total
leukocytic count.
Basophils are motile, short-lived, with a life span of 10 – 15 days. They are
easily seen in Wright- stained blood films because of their coarse large
basophilic granules which hide the S-shaped nucleus.
Basophils have fewer, irregular in size and shape specific granules which
are highly soluble in water, metachromatic with toluidine blue, containing
histamine, heparin, SRS-A and ECF-A, and smaller nonspecific granules.

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Though similar to mast cells in being basophilic, with metachromatic
granules containing histamine and heparin; basophils differ in origin,
granules number, size and shape of nucleus.
Function:
 Under certain circumstances, basophils migrate to connective tissue
to function as mast cells in reactions of immediate hypersensitivity.
 Produce heparin and histamine
 Low grade phagocytosis
 Involved in certain types of inflammation such as cutaneous
basophil hypersensitivity.

II. Agranular leukocytes (Agranulocytes):

These are leukocytes which lack specific granules but they have
nonspecific granules.
1. Lymphocytes: In normal individuals, lymphocytes constitute 20 -40% of
the total leukocytic count. Lymphocytes are small, medium and large size.
The size of small lymphocytes ranges between 6 and 8 um in diameter.
The large lymphocytes, up to 18 um in diameters, with large indented
nuclei are difficult to distinguish from monocytes. However, large
lymphocytes can be differentiated by the condensed, densely stained
heterochromatin and the nonvisiblity of nucleoli in routine stains.
The large lymphocytes are believed to be activated by specific antigens;
they differentiate into effector T or B lymphocytes.

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The small lymphocyte nucleus is nearly spherical or slightly indented
dense, dark and usually surrounded by a rim of basophilic cytoplasm. The
cytoplasm has few mitochondria, small Golgi, centrioles, free ribosomes
and short microvilli seen by EM (more numerous on B lymphocytes than T
lymphocytes).
Some lymphocytes may live few days, others may live for years. They are
the only type that may return to blood after the process of diapedesis.
Function:
 Lymphocytes are responsible for the body humoral defense against
bacterial and viral infection.
 B lymphocytes produce antibodies to resist infection.
 Helper T lymphocytes release substance to activate B lymphocytes.
 Cytotoxic T lymphocytes secrete substance to kill virus infected
cells and foreign cells transplanted from other individuals.

2. Monocytes: These are the largest leukocytes being 12 - 20 um in

diameter and make up to 3 - 8% of the total leukocytic count.
In blood smear, it is often difficult to distinguish between monocytes and
large lymphocytes. Monocytes have the following features:
 Monocyte nucleus is usually indented or horseshoe-shaped
eccentrically located and lighter than lymphocyte nucleus.
 Monocyte cytoplasm is basophilic and contains fine azurophilic
granules which are lysosomes (despite of their name nongranular).

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  Nucleus has one or two nucleoli
 Cytoplasm contains Golgi, RER, polysomes, mitochondria,
lysosomal granules and cytoplasmic vacuoles.
 Cell surface has many microvilli and pinocytotic vesicles.
 They originate from stem cells in the red bone marrow.
 Their average life span in the blood stream is three days.
Function:
 Phagocytic cells of the blood
 Necrotaxis: migrate into tissue and differentiate into tissue
macrophages, engulf and destroy tissue debris and foreign materials.
 Important role in mononuclear phagocytic system (monocytes,
macrophages, multinucleated giant cells, Kupffer cells, microglia,
Langerhans cells, osteoclasts, APCs).

C. Blood platelets (Thrombocytes):

These are ovoid, biconvex cytoplasmic fragments (not cells), 2 - 4 um in
diameter, derived from megakaryocytes, the largest cells of the red bone
marrow.
The platelets number: 200,000 - 400,000 / microliter (cu.mm) of blood;
their life span is about 10 days.
The platelet has a central basophilic staining region called granulomere or
chromomere and a thin pale homogenous peripheral zone called hyalomere.

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Platelets contain an open canalicular system which connects to
invaginations of plasma membrane to facilitate liberation of stored
molecules. A marginal bundle of microtubules lies at the periphery of the
platelets to maintain their ovoid shape. Hyalomere contains a number of
dense irregular tubes called dense tubular system (DTS), actin and myosin
filaments.
Outside the platelet membrane, a cell coat rich in glycosaminoglaycans and
glycoproteins is present and helps in platelet adhesion.
In humans, the central granulomere contains membrane-bound granules of
variable sizes, mitochondria and glycogen particles. The granules are of
three types, alpha (α), (delta) δ-granules or dense bodies and lambda (λ)
granules.
Alpha granules are the largest, more numerous and contain fibrinogen,
platelet-derived growth factor, von Wille-brand`s factor (promotes adhesion
of platelets to endothelial cells) and platelet factor IV which stimulate
blood coagulation.
The dense bodies contain calcium ions, pyrophosphates, ADP, ATP and
serotonin absorbed from the plasma.

In blood film, platelets tend to clump together making it difficult to count.

In order to count the blood platelets properly, it is essential to use
anticoagulant (antiagglutinant) in the syringe or in the pipette used.
The platelets function in blood clotting and controlling hemorrhage as
follows:
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  Primary aggregation, immediately at the damaged tissue forming
platelet plug.
 Secondary aggregation when the platelets release an adhesive
glycoprotein and increase the size of platelet plug.
 Blood coagulation due to release of factors from blood plasma,
fibrin, trapping blood cells and platelets making thrombus or blood
clot.
 Clot retraction due to interaction of platelet actin and myosin and
ATP.
 Clot removal by proteolytic enzyme, plasmin released by
endothelium and enzymes released from λ-granules.
HEMATOPOIESIS
Hematopoiesis or hemopoiesis means formation of blood elements. This is
done by continuous replacement by stem cells located in myeloid tissue or
hemopoietic tissue. In early intrauterine life, blood cells arise from
mesoderm of yolk sac, then from spleen, liver and bone marrow.
After birth, blood elements are derived from stem cells located in red bone
marrow.

All blood cells arise from pluripotential stem cells which are able to renew
themselves and are located in bone marrow. Some of these cells remain as
a reservoir of stem cells and the rest differentiate into two cell lines:
 Lymphoid multipotential cells migrate to lymphoid organs

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  Myeloid multipotential cells which develop in the bone marrow into
granulocytes, monocytes, erythrocytes and megakaryocytes.

Bone Marrow
Bone marrow is one of the largest organs of the body, and the main location
of hemopoiesis. It is a specialized type of connective tissue, derived from
mesenchyme and present as two forms, yellow and red.
 Yellow bone marrow composed of large number of fat cells. It
present in marrow cavities of long bones. Its main function is fat
storage and changes to red bone marrow in severe cases of bleeding
and hypoxia.
 Red or hematogenous bone marrow; appears red due to presence of
blood and blood-forming cells.
In newborns, all bone marrow is red. In adult, the red bone marrow is
presents in the ends of long bones and flat bones such as: sternum,
clavicles, ribs, skull, vertebrae, and pelvic bones.
Red bone marrow is composed of:
1. Stroma is composed of reticular cells and network of reticular fibers.
The matrix contains collagen type I and III fibers, proteoglycans and
glycoproteins, fibronectin, laminin, and hemonectin which bind the
cell receptors for better cell-matrix adhesion.
2. Hematopoietic cords of stem cells and different stages of blood
elements and macrophages lying within stroma and blood sinusoids.

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 3. Sinusoidal capillaries, wide, numerous, fenestrated and supported
externally by discontinuous layer of reticular cells and loose reticular
fibers.
Functions of bone marrow:
 Production of blood cells
 Destruction of RBCs
 Storage of iron in macrophages, resulting from hemoglobin
break down
Erythropoiesis
By this process, differentiation and maturation of RBCs to attain a specific
shape and perform specific functions. Erythropoietin, a hormone secreted
by the kidney, and other factors such as iron, folic acid and vitaminB12 are
important in the process of erythropoiesis. This process passes through the
following stages:
1. Pluripotent hemopoietic stem cells which are large cells with
basophilic cytoplasm, large pale nuclei with prominent nucleoli.
2. Appearance of colonies of cells to produce erythrocytes called ECFC
(erythrocyte colony forming cells).
3. Proerythroblasts, large cells, with large nuclei and basophilic
cytoplasm.
4. Basophilic erythroblasts, which are smaller cells, with smaller darker
nuclei, and basophilic cytoplasm.

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 5. Polychromatophilic erythroblasts, at this stage, polyribosomes
decrease, and hemoglobin starts to appear in cytoplasm causing
several colors to appear in the cell.
The cells and their nuclei become smaller and cytoplasm becomes
acidic.
6. Orthochromatophilic erythroblasts (normoblasts) are small. Their
cytoplasm is acidophilic with small number of polyribosomes.
Nucleus, small pyknotic, peripheral then extruded out of the cell
then engulfed by macrophages.
7. Reticulocyte: does not have nucleus and soon looses polyribosomes,
mitochondria and other enzymes to become mature erythrocytes.
The process of degradation of organelles is not mediated by
lysosomal enzymes, but by a group of ATP-dependant enzymes.
Normally, reticulocytes form about 1% of the circulating RBCs; they
increase in number in hemorrhage and high altitudes.

Granulopoiesis
Formation of granular leukocytes starts with:
1. Pluripotent hemopoietic stem cells in bone marrow, large cells with
large nuclei and basophilic cytoplasm.
2. GCFC (granulaocyte colony forming cells)
3. Myeloblasts: large cells with large nuclei, dispersed chromatin and
prominent nucleoli.

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 4. Promyelocytes, basophilic cytoplasm and azurophilic granules with
lysosomal enzymes (not specific) and myeloperoxidase.
5. Myelocytes start showing signs of differentiation to three types of
granulocytes, regarding size, granules, nuclear shape.
 Neutrophil myeloblasts show neutrophilic specific granules,
smaller size, and multilobed nucleus then becomes mature
neutrophils.
Band cells are immature neutrophils, their appearance in
large number in blood indicates bacterial infection and is
called shift to the left.
 Eosinophil myeloblasts show specific granules and
condensation of nuclear chromatin to give rise to mature
eosinophils.
 Basophil myeloblasts show specific basophil granules and
condensation of chromatin into an S-shape to become
basophils.

Neutrophil compartments:
During granulopoiesis, neutrophils pass through anatomically and
functionally defined compartments, which are:
1. Medullary compartment where mitosis and maturation occur.
2. Medullary storage where large number of mature neutrophils are stored

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 3. The circulating compartment consists of circulating neutrophils in blood
vascular system
4. The marginating compartment consists of non-circulating neutrophils
which are present in blood capillaries and temporarily excluded.

Agranulopoiesis
Formation of lymphocytes and monocytes:
All lymphocytes progenitor cells originate in bone marrow. Some of them
migrate to thymus where they mature into T lymphocytes, then released to
lymphoid organs. Others differentiate into B lymphocytes inside bone
marrow then migrate to lymphoid organs.

Formation of T or B lymphocytes:
1. Pluripotent hemopoietic stem cells, large cells with large nuclei
2. Lymphocyte-forming colony cells
3. Lymphoblasts, large cells capable of dividing two to three times
4. Prolymphocytes, smaller, more condensed chromatin, but no specific
cell surface antigen.
5. Some lymphocytes migrate to thymus to mature and programmed
into T lymphocytes, where these cells synthesize their specific cell
surface receptors which are recognized only by
immunocytochemical techniques. Then released to circulation then
to lymphoid organs.

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 6. Some other cells stay in bone marrow to mature and differentiate
into B lymphocytes, then synthesize the specific cell surface
receptors, then reach the circulation and then to lymphoid organs.
When activated by antigens, B lymphocytes differentiate into plasma
cells.

Formation of monocytes:
1. They differentiate from pluripotent hemopoietic stem cells in red
bone marrow.
2. Monoblasts differentiate into promonocytes
3. Promonocytes are large cells with basophilic cytoplasm and large
indented nuclei with prominent nucleoli
4. Promonocytes divide twice then develop into monocytes with large
amount of RER and well developed Golgi complex, primary
lysosomes and fine azurophilic granules.
5. Mature monocytes enter blood stream, stay in circulation for about 8
hours then enter the connective tissue and become macrophages.

Formation of blood platelets (thrombocytes):

Blood platelets originate from megakaryocytes in red bone marrow as
follow:
1. Pluripotent hemopoietic cells

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2. Megakaryoblasts are large cells 15-50 um in diameter, large ovoid or
kidney-shape polyploid nucleus (DNA 30 times as normal),
numerous nucleoli. Intense basophilic homogenous cytoplasm.
3. Promegakaryocytes
4. Megakaryocyte, in bone marrow, is a giant cell 35-150 um in
diameter, with large irregular lobulated nucleus. The cytoplasm
contains numerous mitochondria and RER, well developed Golgi
complex. Cell membrane has numerous invaginations which ramify
through the cytoplasm giving demarcation membranes for platelets
formation.
5. Blood platelets migrate to blood stream

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 Cartilage
Cartilage is an avascular connective tissue specialized in providing support,
rigidity and some flexibilty. It is composed of ground substance (matrix),
cells and fibers.
Its ground substance is firm and compact consisting of chondro-muco-
protein which stains blue with basic dyes.
Based on the composition of the matrix, there are three types of cartilage,
hyaline (most common), elastic and fibro-cartilage.

Hyaline cartilage
Hyaline cartilage is the most prevalent type. To the naked eye, hyaline
cartilage looks translucent (glass-like); it could occur as discrete pieces or
associated with bone.
Sites and functions:
 at the ends of ribs at their attachments with sternum
 nose
 larynx
 Trachea and bronchi, C-shape cartilage, provides support for the soft
tissues of trachea to help stay patent.
 ends of long bones in joints
 Fetal skeleton.
Hyaline cartilage is composed of:

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  Matrix: ground substance (chondromucoprotein and chondroitin
sulfate), and fibers (collagen type II)
 cells (chondroblasts and chondrocytes)

Perichondrium:
Hyaline cartilage is covered by perichondrium (except the articular), a
nutritive CT capsule, the perichondrium.
The perichondrium is made of two layers, an outer fibrous made of
collagen type I and elastic fibres, fibroblasts and blood vessels, and an inner
cellular chondrogenic layer whose cells are chondroblasts which
differentiate into chondrocytes.
Perichondrium merges gradually via its chondrogenic zone, rich in
chondroblasts, with the cartilage proper. Perichondrium is important for
growth and maintenance of cartilage.

Matrix (Extracellular matrix):

The matrix appears semi-rigid and clear to the naked eye.
In H & E stain, it appears amourphous and basophilic despite the presence
of collagen fibrils (mainly type II) which appear in EM and in LM by silver
stains. In LM, the fibrils are too thin to be resolve and the matrix and the
fibrils have the same refractive indices.
The matrix is composed of proteoglycans and structural glycoproteins.
The proteoglycans are:

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  Sulfated glycosaminoglycans (GAGs): chondroitin sulfate and
keratan sulfate covalently bound to core proteins.
 Non-sulfated glycosaminoglycans: hyaluronic acid associated with
large number of proteoglycans forming proteoglycan aggregates.
Proteoglycans give the matrix its basophilic color and bind water to
give flexibility.
 The structural glycoprotein chondronectin specifically binds to
collagen type II and glycoseaminoglycans, thus initiating attachment
between the matrix and chondrocytes.
The matrix is divided into territorial and extraterritorial (inter-territorial)
regions.
The capsular or territorial matrix is the youngest part of the matrix. It is a
thin zone of darker, basophilic matrix immediately adjacent to the rounded
lacunae; it represents the capsule of the lacunae.
This capsule is not a membrane; it is a basophilic rim of matrix rich in
glucosamino-glycans and poor in collagen. The matrix between the cells or
the groups of cells is called the interterritorial matrix.

Cells of cartilage:
 Chondroblasts: secrete collagen and the matrix
 Chondrocytes: secrete collagen and the matrix; they lie within
lacunae. The lacunae lying adjacent to perichondrium, are small and

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 flattened, while the lacunae deeper within the cartilage are larger and
rounded.
The chondrocytes occupying the flattened lacunae represent young cartilage
cells which have differentiated from the cellular perichondrium. This
represents appositional growth of cartilage.
Chondrocytes may appear in groups of up to 8 cells originating from
mitotic divisions of a single chondrocyte, and later the daughter cells
produce matrix material between themselves forming what is called
isogenous groups. This is the interstitial or internal method of cartilage
growth.
The cytoplasm of chondrocytes appears vacuolated due to loss of glycogen
during fixation; the nuclei appear quite dark and wrinkled.
The functions of chondrocytes are under hormonal control; synthesis of
ground substance is enhanced by growth hormone, thyroxine and
testosterone and slowed by cortisone. Cartilage growth is controlled by
somatotropin which acts on liver cells to synthesize somatomedin C which
in turn acts on cartilage cells to promote their growth.
Nutrition of cartilage:
Cartilage is avascular tissue; it gets its nutrients by diffusion from the blood
vessels in perichondrium.
Growth of cartilage:
Growth of cartilage depends on the growth hormone of pituitary
(hypophysis) which acts on liver cells to synthesize somatomedin C which
in turn acts on cartilage to promote its growth.
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The growth occurs in two ways:
1. Appositional growth, by mitosis of chondroblasts in cellular
perichondrium and adding new layers.
2. Interstitial growth, where growth occurs from inside the cartilage by
mitosis of the young chondrocytes.
Histogenesis of cartilage:
All types of cartilage develop from mesenchymal tissue as follows:
 The branched mesenchymal cells proliferate rapidly by mitotic
division and modify their shape to round cells giving mesenchymal
cell condensation.
 The condensed cells are differentiated in the tissue center into
chondroblasts with basophilic cytoplasm, very rich in ribosomes.
 Chondroblasts start synthesis of matrix and the cells become
separated from each other, and the cells in the center become
chondrocytes ying in lacunae.
 In the developing cartilage, the central cells are chondrocytes while
the peripheral cells are chondroblasts; the superficial layer of
mesenchymal tissue will differentiate into perichondrium.
Degeneration and regeneration of cartilage:
 In old age, hyaline cartilage shows degenerative changes and
calcification of its matrix.
 Regeneration of damaged cartilage takes place by the cellular layer
of perichondrium. In young age regeneration produces new

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 cartilage; while in adults, cartilage regenerates very poorly and heals
by thick scar of dense connective tissue.

Elastic cartilage
Elastic cartilage is similar to hyaline cartilage except for the elastic fibers in
its matrix; it is covered by perichondrium, made of outer fibrous and inner
cellular layers.
Matrix:
The matrix contains collagen fibers type II and elastic fibers which branch
interlace and run in all directions. The elastic fibers diverge around the
lacunae forming a network which provides the cartilage with flexibility.
Although not seen in the section, collagen fibers are present among the
elastic fibers.
Cells: Chondrocytes lie inside lacunae, and form cell nests.
Elastic cartilage is present in:
 Ear pinna
 Eustachian tubes
 Epiglottis
 Some cartilage of larynx
Fibrocartilage
Fibrocartlage has no perichondrium.
Matrix: The matrix is filled with parallel dense collagen fibers type I.
In H&E stain, collagen fibers stain acidophilic.

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Cells:
The chondrocytes lie single or in isogenous groups, in rows between heavy
bundles of collagenous fibers. The lacunae are considerably smaller than in
hyaline or elastic cartilage. It is important to remember that fibrocartilage
always blends into either hyaline cartilage or a dense fibrous connective
tissue. Sites:
 Intervertebral discs
 Pupic symphysis
Function: Resists stretching and provides restricted mobility
Srtucture of intervertebral disc:
Intervertebral disc is composed of:
 A central jelly-like region called nucleus pulposus which contains
collagen type II and it is not a cartilage. Its size decreases by age.
 A fibrous ring-like peripheral region called annulus fibrosus
surrounding the nucleus pulposus; composed of lamellae of
fibrocartilage made of many thick collagen fibers type I with few
chondrocytes in lacunae.
Disc prolapse or herniation:
There is herniation of nucleus pulposus through the torn annulus fibrosus
into the vertebral canal, causing compression on the spinal nerves and
severe pain.

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 BONE
Bone is a hard connective tissue in which the fibers are impregnated with
mineral substances mainly calcium phosphate and calcium carbonate. It is
covered by periosteum and lined by endosteum.
The bone is composed of matrix (containing ground substance and fibers) and
cells. Bone serves for support, attachment, leverage, protection and mineral
storage.

Periosteum:
Periosteum is a connective tissue sheath covering the outside of the bone;
bone interior is lined by endosteum.
Sharpey`s fibers are bundles of periosteal collagen fibers penetrating the
bone matrix and binding periosteum to bone.
Periosteum is composed of two layers:
 Inner cellular layer containg osteoprogenitor cells which are also
present in edosteum.
 Outer fibrous layer rich in collagen fibers, fibroblasts, nerves and
blood vessels.

Functions of periosteum:
1. Periosteum provides muscle attachment
2. It carries blood and nerve supply
3. Its osteogenic layer is important in bone growth and repair

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Endosteum:
Endosteum lines all the internal surfaces of bone cavities. It is very thin and
composed of a single layer of osteogenic cells supported by small amounts of
reticular connective tissue. In bone growth and repair, the osteogenic cells
differentiate into osteoblasts.

Bone Matrix:
The bone matrix is made of calcified bone lamellae; it is composed of organic
(osteoid= immature bone matrix without minerals) and inorganic (minerals)
components.
The organic part includes collagen type I, proteoglycans and glycoproteins:
 Highly ordered collagen fibers, type I which forms 90% of the organic
components
 Proteoglycans
 Bone specific proteins:
1. Osteocalcin: non-collagen organic material binds calcium
during mineralization
2. Osteonectin which serves bridging function between collagen
and minerals.
3. Sialoprotein, rich in sialic acid concentrated from plasma.

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The inorganic, minerals constitute about 50% of the bone dry weight; it is
composed of calcium and phosphorus mainly and small amounts of
carbonates, citrates, magnesiu, potassium and sodium.
 Calcium phosphate, an insoluble salt, forms rod-shaped
hydroxyapatite crystals. A layer of water and ions (hydration shell)
surrounds each hydroxyapatite crystal; this may facilitate the exchange
of ions between crystals and body fluids. Hydroxyapatite has high
affinity to heavy metals and radioactive substances.
 Pyrophosphate inhibits calcium deposition, important for controlling
mineralization of bone.

Bone Cells:
1. Osteogenic or osteoprogenitor cells: these are present in cellular layer
of periosteum and in endosteum; they are stem cells or precursors for
osteoblasts. They are fusiform with elongated pale staining nuclei.
2. Osteoblasts are differentiated cells arising from osteogenic cells; they
are polarized, located at the bone surface as one layer, side by side.
When they are active, during bone formation, they appear large
cuboidal to columnar cells with the ultrastructure of active cells in
protein synthesis for export. Their nuclei are large, vesicular, oval and
eccentric and their cytoplasm is deeply basophilic rich in phosphates,
ribosomes, RER, mitochondria and well-developed Golgi complex.
When they are less active, they attain flattened shape and less
basophilic cytoplasm, but they revert to the active state as needed.
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 When they become enveloped or trapped by their own secretion,
they transform into quiescent mature bone cells, osteocytes, lying
inside lacunae deeper in the matrix.
The functions of osteoblasts are:
 To form the bone non-calcified osteoid part of the matrix,
glycoproeins and proteoglycans.
 Synthesize type I collagen.
 Osteoblasts have surface receptors for parathyroid hormone
which activates the cells to produce the osteoclast stimulating
factor, cytokine.

3. Osteocytes are the main cells of adult bone; they are long-living cells
that reside within bony lacunae between the matrix lamellae, one
osteocyte in each lacuna.
Osteocytes are flat, almond-shape cells, smaller with darker nuclei
than osteoblasts. The cytoplasm is less basophilic, with less RER and
numerous long cytoplasmic processes. The processes are running in
bony canaliculi radiating from lacunae. The ends of the processes of
neighboring osteocytes meet within canaliculi and have gap junctions
at the site of contact through which osteocytes exchange molecules
and get nourished from blood capillaries.
Their main function is to maintain the bone matrix.

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4. Osteoclasts are multinucleated (5 - 50 nuclei) giant, branched, motile
cells. They are often located in shallow bony depressions caused by
enzymatic resorption, called Howship's lacunae.
Osteoclast cytoplasm is acidophilic, rich in lysosomes, mitochondria
and well-developed Golgi complex. When the cell is active, its side
facing the bone matrix is folded forming ruffled border.
Osteoclasts are derived from fusion of bone marrow-derived
mononucleated cells and they belong to mononuclear phagocyte
system. Between the bone and the ruffled surface of osteoclast, there
is a clear zone of cytoplasm, microenvironment, devoid of organelles
but rich in actin filaments (podosomes) serving as an attachment site
for osteclasts to the bone matrix to create a sealed environment for
bone resorption.
Osteoclasts secrete collagenase and other enzymes (cathepsin), and
pump protons into the zone between ruffles and the matrix lowering
the pH to suit lysosomal enzymes and help dissolve calcium crystals.
Osteoclasts are under the control of cytokines and calcitonin hormone
to which they have receptors but not for parathyroid hormones.
Their functions:
 Bone resorption
 Remodeling and shaping of bone
 Phagocytosis of damaged bone.

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Types of bone:
I. According to bone development and microscopic examination, there are
two types of bony tissue:
1. The Primary (immature or woven) bone tissue:
It is the first bone to develop in embryonic life and in repair after fractures
and bone damage. It has random deposition of coarse collagen fibers, lower
mineral contents and higher proportion of osteocytes than the secondary type.
It is temporary and is replaced by secondary bone in adults except in some
places near the sutures of the skull, tooth sockets and insertions of some
tendons.
2. The secondary bone tissue (lamellar or mature):
This type occurs in adults, where collagen fibers are arranged in lamellae,
parallel to each other and concentrically arranged around a vascular canal.

II. According to the size of spaces in the bone and its trabeculae, it is divided
into:
 Spongy
 Compact

Bone Preparation for Histological Examination:

Bone is prepared in two ways to be examined by the microscope.
a. Ground bone: small pieces of dry bone to be ground until become very
thin and transparent, and then mounted on histology slides without
staining. In this method, all organic substance and cells are dead. This
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 is useful to demonstrate lacunae, canaliculi, lamellae, haversian canals
and Volkmann`s canals
b. Decalcified bone: In this method, calcium salts are removed by acid
solutions like nitric acid. The bone becomes soft, prepared like other
organs then cut into sections and stained.

Anatomical parts of bone:

 Epiphysis: the bulbous end of a long bone, composed of spongy
covered with a thin layer of compact bone.
 Diaphysis: the cylindrical part of a long bone made of compact bone
with very small part of spongy bone around the bone marrow cavity.
 Plates or tables of skull made of compact bone.
 Diploё: spongy bones between the two tables of skull.

Compact Bone
Compct bone is thick solid mass present in the shaft of long bone and
covering the flat and irregular bones. It is covered by periosteum and its
cavity is lined with endosteum. It is composed of cylindrical subunits called
Haversian systems or osteons.

Osteons or Haversian systems:

An osteon consists of a central canal surrounded by concentric bone lamellae
and lacunae. The central canals run longitudinally in the bone and serve as

151
channels for blood vessels and nerves; they are lined by endosteum which is
not present in dried ground section.
The concentric bone lamellae are successive layers of bone which surround
the Haversian canal. Within the concentric lamellae of an osteon, lacunae are
seen connected to each other and to central canals by canaliculi. In living
state, osteocytes occupy these lacunae and their processes occupy the
canaleculi.
In dried ground sections, lacunae appear dark due to presence of trapped air
inside them. Cement lines are refractile lines which limit the boundaries of
each osteon.
Interstitial lamellae are irregular lamellae lying between haversian systems.
External circumferential lamellae are bone lamellae running close and
parallel to periosteum on the outer surface of bone.

Internal circumferential lamellae are bone lamellae lying close to

endosteum at the inner surface of bone.

Volkmann`s canal are transverse or oblique canals connecting haversian

canals together and to periosteum and marrow cavities. They are lined with
osteogenic cells and contain blood vessels.

Sharpey`s fibers are perforating collagen fibers connecting the fibrous layer
of periosteum to haversian systems.

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 Spongy (cancellous) Bone
Spongy bone is made of branching and interconnecting trabeculae with large
marrow spaces in between. There are no Haversian systems and the lamellae
are irregularly arranged.
The marrow spaces are full of active red bone marrow and lined with
endosteum. The lining is difficult to identify since it is very thin and consists
thin layer of osteogenic cells and reticular connective tissue.
The osteocytes are occupying the lacunae, whereas osteoblasts are occupying
the surface of trabeculae. Spongy bone is found at the ends of long bones and
in the center of flat and irregular bones.

Bone Ossification
Bone ossification is the process of bone formation; it starts in mesenchymal
tissue of the embryo by two ways:
 Intramembranous ossification where the bone matrix is secreted by
osteoblasts in mesenchymal membranes.
 Endochondoral or intracartilagenous ossification: by deposition of
bone matrix on a pre-existing cartilage model.
In both methods of ossification, the first bone to appear is primary and soon
replaced by secondary. Different stages of bone development appear side by
side.

Intramembranous ossification:

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By this method, the membranes of mesenchymal connective tissue are
transformed into spongy bone. This process occurs in flat bones (skull),
growth of short bones and thickening of long bones. It occurs as follows:
 The cells of mesenchymal connective tissue condensation divide and
differentiate into osteogenic cells which increase in size and
differentiate into osteoblasts. This starting point of ossification is
called primary ossification center.
 The osteoblasts synthesize the organic component of bone matrix,
osteoid, and the alkaline phosphatase.
 Deposition of calcium salts in the matrix results in trapping of
osteoblasts inside lacunae to become osteocytes. The scattered
developing bone areas are called spicules which appear as elongated
structures with cavities containing blood capillaries, bone marrow
cells and undifferentiated cells.
 The spicules fuse with each other giving rise to spongy bone.
 The remaining mesenchymal tissue between the specules is penetrated
by blood vessels and differentiated into red bone marrow.
 Several ossification centers fuse together and replace the original
connective tissue.
 The mesenchymal tissue at the surface of the membranes becomes
periosteum and endosteum.

Note that:

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  The fontanelles of newborn infants are non-ossified connective tissue
of the skull.
 The active metabolites of Vitamin D3 are involved in particular in
stimulating dietary calcium absorption through the small intestine.
Lack of vitamin D3 can result in improper calcification of bone tissue
and the development of rickets.

Endochondoral (intracartilagenous) ossification:

This type of ossification takes place in a cartilage model; it is responsible for
bone formation in long and short bones. It occurs in two phases:
1. Hypertrophy and destruction of chondrocytes in a cartilage model
leaving expanded spaces separated by septa of calcified cartilage.
2. Formation of osteogenic bud consisting of ostegenic cells and blood
capillaries invading the spaces left by degenerated chondrocytes.
The fetal skeleton including long bones is made of hyaline cartilage.
Each long bone is composed of a cylindrical shaft (diaphysis) and two
dilatations (epiphyses), one at each end of the shaft.

Formation of long bones takes place as follows:

1. The first step is the formation of bone by means of
intramembranous ossification in perichondrium surrounding hyaline
cartilage of diaphysis. Thus producing the bone collar in the deep
part of perichondrium.

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 2. The perichondrium opposite the bone collar is changed to
periosteum, because it covers the newly formed bone.
3. Chondrocytes in the cartilage model, begin to degenerate since
nutrient diffusion is prevented by the bone collar.
4. Calcium salts start to deposit in the cartilage matrix which become
calcified.
5. Periosteal blood vessels (osteogenic bud) and osteogenic cells
invade the spaces in calcified cartilage after the death of
chondrocytes.
6. Osteogenic cells proliferate to give osteoblasts which arrange
themselves along the marrow spaces and start bone matrix
formation.
7. Spongy bone will be formed in the center of diaphysis which is
called primary ossification center.
8. Marrow spaces fuse together forming a single bone marrow cavity
in the center of diaphysis.
9. A secondary ossification center arises later in embryonic
development, where the growth will be radially rather than
longitudinally.

The epiphyseal plate is made of hyaline cartilage and connects the diaphysis
to epiphysis. It is continuously replaced by bone matrix from secondary
ossification center.

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The central marrow cavity is the oldest part of developing bone.
During endochondoral ossification, the epiphyseal cartilage is divided into 5
zones:
1. The zone of reserve or resting cartilage which consists of typical
hyaline cartilage without morphologic changes. It occupies a large
area of epiphyseal plate.
2. The zone of proliferation, where the chondrocytes divide rapidly and
form columns or rows of stacked cells parallel to the long axis of bone.
3. The zone of maturation and hypertrophy, in which the cells and
lacunae are enlarged. The chondrocytes no longer divide; they contain
large amounts of glycogen and alkaline phosphatase.
4. The zone of calcification, which is a narrow zone of calcified cartilage
in which the matrix stains more basophilic or slightly darker than the
preceding zone. In fact, it is difficult to distinguish this zone from the
zone of hypertrophy, and sometimes considered the same zone.
5. The zone of ossification, where the calcified cartilage is resorbed and
replaced by bony tissue. Enlarged lacunae and spaces are invaded by
blood vessels and osteoprogenitor cells which differentiate into
osteoblasts, start the bone formation.
Bone Remodeling:
Remodeling of bone is carried out by phagocytic cells derived from blood
monocytes called osteoclasts. Osteoclasts are rich in lysosomes; they secrete
collaginase enzymes. As osteoclasts resorb bone, new bone is laid down by
osteoblasts.
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Growth of bone:
Bone growth occurs in two ways:
 Interstitial growth by increasing the length through ossification of
epiphyseal plates.
 Growth in width by increasing the diameter of long bones due to
continuous bone deposition under the periosteum.
Effect of hormones on bone growth:
1. Growth hormone stimulates proliferation and maturation of
chondrocytes in epiphyseal plate. Excess of the hormone leads to
gigantism (young) and acromegally (adults) while deficiency leads to
dwarfism.
2. Diminshed gonadal hormones in aging disturb the balance between
bone deposition and bone resorption leading to fragile bones.
3. Parathyroid hormones increase the number of osteoclasts promoting
bone resorption and increasing blood calcium levels.
4. Calcitonin hormone of thyroid parafollicular cells inhibits osteoclast
bone resorption, thus lowering blood calcium levels.
Bone Fracture Repair:
1. At the site of fracture, there is destruction of bone cells and matrix.
2. The damaged blood vessels produce blood clot.
3. During repair, the blood clot, the damaged bone cells and matrix are
removed by macrophages.

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 4. The osteogenic cells of periosteum and endosteum proliferate
intensely, forming cellular tissue around the fracture and extending
between the two ends of fracture.
5. A small fragment of cartilage is formed in the connective tissue of
fracture forming temporary callus.
6. The osteogenic cells are reactivated to form bony callus
(intramembranous) to replace the temporary callus after resorption.

Joints
Joints are regions where bones are joined together. There are two major
groups of joints, synarthroses and diarthroses.
Synarthroses:
In these types of joints, there is no or very limited bone movements. These
are divided into three types:
1. Synostosis (bony joints):
Bones are united by bony tissue (skull sutures); or by interosseous ligaments.
In this type of joints, there is very limited or no movements.
2. Synchondrosis:
Bones are joined by hyaline cartilage such as epiphyseal plate and the joint
between the first rib and sternum.
3. Syndesmosis:
This type permits certain amount of movement. The bones are joined by
interosseous ligaments (pubic symphysis, intervertebral joints).
Diarthroses:
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These are joints of long bones permitting great mobility such as elbow and
knee joints. They have ligaments and capsule to maintain the bone contact.
The capsule encloses articular cavity which contains clear viscous, synovial
fluid, which is a transudate of blood plasma with high concentration of
hyaluronic acid.
The ends of the bone are covered with articular cartilage (hyaline) with no
perichondrium. The articular capsule has two layers, an external fibrous
layer and an inner synovial layer.
The outer fibrous layer is made of dense connective tissue; it encloses the
ligaments and the tendons inserted near the joints.
The inner synovial layer of internal surface is lined by squamous or cuboidal
cells with thin layer of loose connective tissue underneath.
EM examination shows that, there are two types of cells in the synovial
membrane: one type is phagocytic and the other cell type resembles
fibroblasts.
Clinical Notes:
1. Calcium deficiency in children causes rickets (slowly growing
deformed bones).
2. Calcium deficiency in adults gives rise to osteomalacia, deficient
calcification of newly formed bones and delacification of already
calcified matrix.
3. Osteoporosis occurrs in females after menopause due to decreased
bone formation

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 Muscle
Muscle or muscular tissue is one of the four basic tissues. It is highly
specialized for contractility.
The contraction occurs when two types of filaments, actin and myosin
interact with each other and interdigitate over a large portion of their lengths,
and thus pulling the two ends of the fibers closer together.
There are three types of muscle:
 Smooth muscles.
 Skeletal muscles.
 Cardiac muscles.
The cardiac and skeletal muscles are striated due to presence of
microscopically cross striations on the fibrils. Only skeletal muscle is
voluntary.
Muscular tissue has its own characteristic terms such as:
Plasmalemma = Sarcolemma
Cytoplasm = Sarcoplasm
Endoplasmic reticulum = Sarcoplasmic reticulum
Mitochondria = Sarcosomes

Smooth Muscles
Smooth muscles are involuntary, non-striated, and present in the wall of the
viscera (hence called visceral muscles), like intestine, ureters, urinary

161
bladder, uterus, and blood vessels. They are controlled by the autonomic
nervous system.
In LM examination:
 Smooth muscles appear as long, non-striated, fusiform cells with
single, oval nuclei situated in the central wide part of the cell. The
nuclei appear wrinkled in the contracted muscles.
 Smooth muscle cells vary in length from 20 um in blood vessels to
500um in pregnant uterus.
 Each smooth muscle cell is enclosed by sarcolemma, a basal lamina
surrounded externally by a network of reticular fibers that binds
cellular units together to produce coordinated contraction force and
carry blood vessels and nerves.
 Smooth muscle cells are connected to each other via gap junctions.
By EM examination:
 Smooth muscle cells contain on both sides of the nucleus, Golgi
complex, mitochondria, RER, poorly developed sarcoplasmic
reticulum but no T tubules.
 Pinocytotic vesicles are present near the cell surface. Smooth muscle
cells are very rich in intermediate filaments, desmin in all smooth
muscles and vimentin is vascular smooth muscles.
 The sarcoplasm contains actin, tropomyosin and myosin filaments
arranged in a crisscross oblique manner forming fine network.

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  All filaments terminate in the dense bodies (cytoplasmic and
membrane-associated) which contain actin binding-proteins, α-actinin
and vinculin; these proteins maintain the longitudinal orientation of
the contractile filaments.
 Dense bodies transmit the contractile force to the adjacent smooth
muscle cells. The flask-shape infolding of plasma membrane are
called caveolae, they correspond to T tubules.

Contraction of smooth muscle occurs by sliding filament mechanism:

 Contraction is initiated by influx of Ca²⁺ and regulated by hormones
which act via cAMP.
 Calcium ions bind a calcium binding protein, calmodulin which
activates the myosin light-chain kinase, the enzyme responsible for
myosin phosphorylation.
 After phosphorylation, myosin interacts with actin resulting in muscle
contraction. Estrogen increases the cAMP which activates myosin
phosphorylation promoting muscle contraction. When cAMP
decreases by the effect of progesterone, smooth muscle relaxation will
occur.
Smooth muscles are present in the walls of viscera, like intestine, ureters,
urinary bladder, uterus, and blood vessels.
Innervarvation: smooth muscles are innervated by autonomic nervous
system (sympathetic and parasympathetic). They receive adrenergic and

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cholinergic nerve endings. The nerve supply modifies and does not initiate
the muscle activity.
Functions of smooth muscle:
 Regulation of lumen sizes in hollow organs
 Rhythmic contractions of GIT
 Synthesis of collagen, elastin and proteoglycans.

Skeletal Muscles
Skeletal muscles are composed of long, cylindrical, multinucleated
(syncytium) muscle fibers, formed by fusion of multiple myoblasts during
early embryonic life.
The skeletal muscle is surrounded by a thin layer of dense connective tissue
called epimysium. From the epimysium, connective tissue septa extend
inward to surround each of the muscle fasicles forming the perimysium, from
which, delicate network of reticular fibers envelop the individual muscle
fibers forming the endomysium.
The connective tissue septa carry blood vessels, nerves and lymphatics and
help transmit the mechanical force of contraction.
Skeletal muscles have very rich capillary network made of continous
capillaries.
The muscle plasmalemma (cell membrane) is called sarcolemma and its
cytoplasm is called sarcoplasm.
By light microscopic (LM) examination:

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  In longitudinal section, the muscle fibers are long cylindrical,
multinucleated with peripherally located nuclei.
 The muscle fibers are composed of bundles of subunits called
myofibrils running in parallel plane to one another and to the axis of
muscle fibers.
 The arrangement of contractile proteins in the sarcoplasm gives the
cross striations, alternating dark (A) and light (I) bands.
EM examination of muscle fibers shows:
 The sarcoplasm is filled with long cylindrical bundles of myofibrils
which are composed of bundles of myofilaments running parallel to
the long axis of the fibrils.
 There are two types of myofilaments, thin (actin) and thick (myosin)
filaments.
 Each I band is bisected by a dark transverse Z line or disk.

The segment between two successive Z lines is called sarcomere.

Sarcomere is the smallest contractile unit of skeletal muscle extending
between two Z lines; it includes an A-band and half of the two adjacent I-
bands.
The thick filaments occupy the central part of sarcomere, the A-band; while
thin filaments run parallel to, and in between the thick filaments and one of
their ends are attached to Z line.

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A band (anisotropic), broad and dark, consists mainly of thick filaments plus
some overlapping thin filaments. It remains constant in width during
contraction.
I band (isotropic), broad and light, consists only of thin filaments. It becomes
narrower during contraction.
Z line bisects the I band; Z lines are drawn together during contraction. It is
seen better at electron microscopic (EM) level.
H band (Hensen`s) is a lightly-stained region lying in the middle of A band in
resting muscles; it is composed only of thick filaments. H band is greatly
reduced in width during contraction state.
M line is a light band lying in the center of H band; it is the site of lateral
connection between the adjacent thick filaments.
At the site of overlapping of thick and thin filaments, one thick filament is
surrounded by six thin filaments in the form of hexagon.
The thin filaments are composed of the proteins, actin, troponin and
tropomyosin while thick filaments contain the protein myosin.

Muscle cells are rich in mitochondria, glycogen, myoglobin (oxygen storage

molecule) and Golgi complex.

Transverse (T) tubules:

These are tubular, finger-like invaginations of the sarcolemma that extend
deep into the substance of the muscle fibers. They form anastomosing
network of tubules that encircle the A-I junction of each sarcomere in every
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myofibril. They enter the fiber at the junction of A and I bands and branch
within the fiber in a transverse plane.
The main function of T tubules is to conduct the wave of depolarization into
all parts of the muscle fiber, hence provide a unified contraction. T tubules
are closely associated with sarcoplasmic reticulum.

Sarcoplasmic reticulum:
In skeletal muscles, the sarcoplasmic reticulum (SER) forms a unique self-
contained compartment of cytoplasm composed of flattened cisternae and
tubules surrounding each myofibril like a collar. On either side of each T
tubule, the sarcoplasmic reticulum expands forming terminal cisternae, one
on A band and one on I band. The function of sarcoplasmic reticulum is to
regulate the concentration of calcium ions within myofibrils.

Triad:
Each pair of terminal cisternae together with a T tubule forms a triad near the
junction of A and I bands of each sarcomere. Its main function is to transmit
the wave of depolarization to sarcoplasmic reticulum.

Types of skeletal muscle fibers: red, white and intermediate.

1. Red muscle fibers (type I): These are slow twitch fibers; they are
highly vascular and contain large amount of myoglobin, cytochromes,
mitochondria (aerobic metabolism), ATP and succenic dehydrogenase.
They are smaller in diameter than the white; they have slow continuous
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 contraction with less fatigability; they get their energy from oxidative
phosphorylation of fatty acids.
2. White muscle fibers (type II): These fibers have fast discontinuous
contraction; they have larger diameter, poor blood supply and contain less
myoglobin and cytochromes and fewer mitochondria (anaerobic
metabolism), rich in glycogen and glycolytic enzymes. They contract fast
and get fatigued quickly. There are 3 subtypes: type IIA, IIB (the fastest)
and IIC.
3. Intermediate fibers are of intermediate size and their contents of
myoglobin, mitochondria and cytochromes are between the previous two
types.

Components of myofibrils: myofibrils are made of thin (actin) and thick

(myosin) filaments
A thin filament is composed of 3 proteins: actin, tropomyosin and troponin.
 Actin is composed of F-actin, made of two strands of globular G-actin
twisted around each other in a helical structure. The actin filaments
are anchored to the protein α-actinin of the Z line. G-actin has binding
sites for myosin. [α-actinin and desmin help to keep myofibrils in
place].
 Tropomyosin molecule is thin, filamentous, running along the groove
of actin helix. It is bound to one troponin molecule and spans 7 G-
actin molecules.

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  Troponin molecule has a specific site on tropomyosin molecule and is
composed of three subunits: TnT binds to tropomyosin, TnC binds to
calcium ions, TnI inhibits actin-myosin interaction.

A thick filament is composed of the protein myosin only.

 Myosin molecule is a large complex molecule consisting of two light
chains of meromyosin and and two twisted heavy chains of
meromyosin with globular head at one end. The heads have binding
sites for actin and ATP.

Mechanism of contraction
The sliding filament theory:
In the resting straietd muscles, the thin and thick filaments are partially
overlapping. Both thin and thick filaments maintain their original length
during contraction. Striated muscle contraction depends on the sliding of thin
actin filaments deep into the A band, thus shortening the sarcomeres. High
concentration of calcium and ATP initiate sliding of thick and thin filaments
over one another.
During relaxation:
 Calcium ions are not available and hydrolysis of ATP by ATPase
enzyme into enrgy and ADP is very slow.

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  Myosin heads cannot interact with actin because troponin-tropomyosin
complex hides the myosin binding sites on G-actin molecules (on the
F-actin filaments).

During contraction:
 Sarcoplasmic reticulum releases calcium ions, which bind to troponin
TnC.
 Binding calcium to troponin will move tropomyosin molecules deep
into the groove of actin helix, thus exposing myosin binding sites and
allowing myosin-actin interaction between the myosin heads and G-
actin molecules.
 As a result of myosin-actin interaction, ATP molecules break down to
ADP and phosphorus ions releasing energy.
 The myosin heads bend and move and pull the actin molecules, thus
initiating sliding movement of actin, and the thin filaments are drawn
further into the A band.
 This process is repeated many times during a single contraction
leading to complete overlapping of actin and myosin filaments and
shortening of the whole muscle. The actin-myosin separates when
myosin binds a new molecule of ATP. In the absence of ATP, in case
of death, actin-myosin complex become stable and leads to sustained
muscle contraction and rigidity, a condition known as rigor mortis.
 I bands, sarcomeres and fibers as a whole become shortened.

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  When Ca²⁺ are removed (taken by SR), the troponin-tropomyosin
complex covers the myosin –binding sites and inhibits actin-myosin
interaction, thus relaxation takes place.

Repair and Healing:

Myofibers have limited regenerative abilities. After injury (trauma) of
skeletal muscle, if the basal lamina stays intact, the satellite cells differentiate
into myoblasts which fuse within the basal lamina to form myotubes which
mature later on to muscle fibers. On the other hand, injury of the basal
lamina will lead to activation of fibroblasts to produce connective tissue
fibers and repair the injured portion of the muscle by scar tissue.

Innervation of skeletal muscles:

A. Motor innervation comes to skeletal muscle fibers through myoneural
junction or motor end plate.
Near the motor end plate, the myelinated axon loses its myelin sheath and
branches to terminate on the muscle fibers forming a dilated termination
(terminal bouton). The terminal axon is covered by Schwann cell cytoplasm.
The terminal part of the axon contains numerous mitochondria and synaptic
vesicles containing neurotransmitter, acetylcholine.
Between the plasmalemma of the axon terminals and the muscle cell
membrane is a space called synaptic cleft containing the amorphous matrix of
basal lamina. At the junction, the axolemma (of axon) is called the

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presynaptic membrane and the sarcolemma (of muscle) is called the
postsynaptic membrane. The post-synaptic membrane (sarcolemma) is
thrown into deep junctional folds where the sarcoplasm contains numerous
nuclei, mitochondris, ribosomes and glycogen granules.
The motor unit consists of a single nerve fiber and all the muscle fibers it
innervates.
Denervation of muscle leads to atrophy and paralysis.

Clinical notes:
In myasthenia gravis, there is a progressive muscle weakness due to
decreased number of acetylecholine receptors at the myoneural junction.

B. Sensory innervation:
Skeletal muscle is supplied by afferent (sensory) nerve endings sensitive to
stretch located in tendons or muscle fibers (proprioceptive
mechanoreceptors). They are present within groups of specialized, spindle-
shaped muscle fibers or tendons called muscle spindles or Golgi tendon
organs.
The muscle spindles consist of modified striated muscle fibers and their
associated nerve fibers; they are most numerous in muscles of extremities and
least in extraocular muscles.
Muscle fibers lying inside the spindles are called intrafusal; they are smaller
than the ordinary muscle fibers and innervated by gamma efferent fibers
(motor).
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The spindles fibers are two types:
a. The nuclear bag fibers containing a group of nuclei in the
expanded mid region, with an afferent primary nerve ending coiled
around the midregion (Annulo-spiral ending).
b. The nuclear chain in which the nuclei are arranged in a row or
chain with a primary nerve ending (annulospiral) coiled around the
midregion plus two secondary endings, one on each side, called flower
spray endings.
The main function of muscle spindle is to detect the changes in muscle
tension and to convey the information to CNS (spinal cord); hence they are
important in maintaining the posture and regulation of the voluntary
movements.

The Golgi tendon organs are proprioceptive sensory nerve endings present
in tendons near their insertions where sensory nerves penetrate large
encapsulated bundles of collagen fibers. They detect the tension changes in
tendons and help to regulate the effort needed to conduct specific movement.

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 Cardiac Muscle
Cardiac muscle is the muscles of the heart; it develops from splanchnic
mesoderm. The muscle fibers are striated and involuntary.
Cardiac muscle is characterized by the following:
 The cardiac muscle fibers run spirally, within the walls of the heart.
 The cardiac muscle fibers appear enclosed by loose connective tissue
called endomysium containing numerous blood capillaries (high
vascularity).
 The fibers appear branching and anastomosing.
 The fibers show cross striations identical to those of skeletal muscles
 Each cardiac muscle cell has one (sometimes two) large, oval,
centrally located nucleus with a clear area at either ends. The atrial
muscle cells are smaller than the ventricular.
 Intercalated discs extend either straight across the fibers or in a step-
wise pattern at irregular intervals.
 The sarcoplasm contains the organelles: myofilaments, mitochondria,
Golgi complex and sarcoplasmic reticulum.
 Sarcoplasm contains the inclusions: myoglobin, glycogen (high
content) and lipofuscin.
 Cardiac T tubules are found at the level of Z band rather than at A-I
junction as in skeletal muscles.

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  Cardiac muscle has diads composed of one T tubules and one cisterna
of sarcoplasmic reticulum. Their number is less in cardiac atrial
muscle fibers.
 Cardia muscle fibers are rich in mitochondria occupying about 40% of
cytoplasmic volume compared to 2% in skeletal muscle.
 Atrial muscle cells contain numerous membrane-limited granules
containing the hormone atrial natriuretic factor (auriculin or
atriopeptin). This hormone acts on the kidney and causes sodium and
water loss, opposing the action of aldosterone and antidiuretic
hormones.

Intercalated disks:
 They are unique distinguishing characteristics to cardiac muscles.
 They are transverse darkly stained lines crossing the muscle fibers at
irregular intervals.
 They represent the junctional complexes between the adjacent cardiac
muscle cells.
 They might be straight or step-like pattern composed of transverse and
lateral parts.
 There are three types of junctions within the disk.
1. Fascia adherents (resemble zonula adherens of epithelium), the most
common, lies in the transverse part; it serves as anchoring (insertion)

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 sites for actin filaments for the terminal sarcomere and represents a
hemi-Z band. It transmits contractility to the adjacent cell.
2. Desmosomes lie in the transverse part and bind the cardiac cells
together to prevent their pulling apart during contraction, and act as
anchoring sites for intermediate filaments.
3. Gap (nexus) junctions lying on the lateral sides of the disk, to provide
ionic continuity between the adjacent cells allowing contractile signals
to pass from cell to cell.

Cardiac conducting system:

The conducting system of the heart is composed of:
 Sinoatrial (S-A) node
 Atrioventricular (A-V) node
 Atrioventricular (A-V) bundle
 Purkinje fibers.

Purkinje fibers (cells) are modified, specialized cardiac muscle fibers; they
are larger than the ventricular muscle fibers, lightly stained and contain fewer
myofibrils which are limited to the periphery of the fiber. They have more
sarcoplasmic spaces surrounding each nucleus, full of glycogen. Nuclei are
rounded and often appear irregular.

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Purkinje fibers extend in the interventricular septum to the papillary muscles
and the lateral wall of the ventricles supplying the apex of the heart before
they proceed to its base.

Hypertrophy, injury and regeneration of cardiac tissue

The cardiac muscle fibers do not regenerate. Any injury of cardiac muscle
cells as in myocardial infarction will result in death of cardiac muscle cells.
Cardiac muscle injury is repaired by fibrous connective tissue (fibrosis). As a
result of the infarct, the cardiac functions at the site of fibrosis are lost, and
the remaining heart muscles undergo compensatory hypertrophy.

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 NERVOUS TISSUE
Nervous tissue is highly specialized for the properties of irritability and
conductivity.
It is organized to form the nervous system which is divided into two major
divisions, the central and the peripheral nervous system.

The central nervous system (CNS) develops from the neural tube ectoderm;
it consists of:
 Cerebrum
 Cerebellum
 Brain stem
 Spinal cord

The peripheral nervous system (PNS) develops from the ectoderm of neural
crest; it consists of all the nervous tissue lying outside of the brain and spinal
cord. It consists of:
 12 pairs of cranial nerves
 31 pairs of spinal nerves
 Ganglia
 Nerve endings
The nerves of the peripheral nervous system are widely distributed
throughout the body.

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Other derivatives of neural crest are chromaffin cells of adrenal medulla,
melanocytes of skin, odontoblasts, cells of pia and arachnoid, neurons of
cranial and dorsal root ganglia, postganglionic neurons of autonomic
(sympathetic and parasympathetic) ganglia, Schwann and satellite cells of
peripheral ganglia.
The autonomic nervous system is composed of sympathetic and
parasympathetic components. This system gives efferent innervation to
viscera, circulatory system and to exocrine glands.

The neuron is the structural and functional unit of the nervous system; it is
defined as the nerve cell body and all of its processes. Nerve cell bodies are
located within the CNS and in the various ganglia outside the CNS. The
processes are dendrites and axons.
Neurons are excitable, meaning that, they respond to external stimuli and
change the electrical potentials of their cell membrane.
Structurally, neurons are classified as multipolar, bipolar and unipolar
according to the number of processes arising from the cell body.

A nerve is an aggregation of the nerve fibers outside the CNS, surrounded by

a sheath of connective tissue. A nerve fiber consists of an axonal process
surrounded by myelin sheath and or Schwann sheath.
A tract is an aggregation of the nerve fibers inside the CNS.
A ganglion is a group of nerve cell bodies located outside the CNS. These
ganglia are cranial ganglia, dorsal root ganglia, sympathetic ganglia and
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parasympathetic ganglia. A group of nerve cell bodies located inside the
CNS is called a nucleus.
Neurons
A neuron is the structural and functional unit of the nervous system which
consists of the nerve cell body and all its processes (the axon and the
dendrites). The neurons are classified on the structural, size and functional
bases.

A. According to number of processes:

1. Unipolar neurons have only one process, e.g. neuroblasts.
2. Pseudounipolar neurons have one process which branch shortly into two
processes, e.g. cells of dorsal-root ganglia and cranial ganglia.
3. Bipolar neurons have two processes, e.g. cells of retina, olfactory mucosa
and spiral ganglia (receptors of sight, smell and balance).
4. Multipolar neurons have many processes; they are the most common,
present in the anterior horn of spinal cord, pyramidal cells of cerebral cortex
and Purkinje cells of the cerebellum.

B. According to function:
1. Motor neurons in anterior horns of spinal cord, which carry motor
impulses from CNS to organs.
2. Sensory neurons which receive sensory impulses from different organs
and send those impulses to CNS, e.g. sensory neurons of spinal ganglia.

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3. Interneurons which act as a connection between motor and sensory
neurons.

C. According to size:
1. Golgi type I, these are large neurons, called principal cells or projecting
neurons; they have long axons carrying information from one part to other
parts of CNS.
2. Golgi type II, they are local circuit nonprojecting neurons, also called
interneurons; they are present in large numbers in CNS.

A neuron consists of cell body (perikaryon), an axon and dendrites.

Perikaryon (soma or cell body):
It is the part of the neuron cotaining the nucleus and cytoplasm. It acts as a
trophic center for the nerve cell, as well as receiving stimuli.
Nucleus:
The nucleus is usually large, rounded, euchromatic with prominent nucleolus.
Binucleated neurons might be seen in sympathetic and sensory ganglia.
RER:
The nerve cell body is rich in RER and polyribosomes which synthesize
structural proteins and proteins for transport. Basophilic aggregates of RER
and free ribosome (polysomes) appear in LM as basophilic granules called
Nissl bodies. Large neurons such as motor neurons contain large number of
Nissl granules.

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Golgi complex: It is present only in the perikaryon; it is composed of several
parallel arrays of cisternae, vacuoles and transfer vesicles. It is important in
packaging of neurotransmitters.
Mitochondria:
Mitochondria are present in perikaryon and in large number especially in the
axon, axon terminals and dendrites.
Neurofibrils or neurofilaments:
These are collectively called neurofibrils, stain brown to black with silver
stains (argyrophilic); they are composed of:
 Neurofilaments (intermediate filaments), provide structural support
 Microfilaments (thin actin filaments) are important in moving the
vesicles towards the cell membrane.
 Neurotubules (microtubules): important in axonal transport of
neurotransmitters and enzymes

The neurofibrils are present in all parts of the neuron. Microtubules of the
perikaryon are in continuous turning over. They are important in
cytoskeleton and cellular transport.
The microtubules of the axon are especially important because they help in
transport of secretory vesicles to nerve endings.
In Alzheimer disease, in aged people, there is accumulation and then
degeneration of neurofilaments in perikaryon of neurons with subsequent loss
of memory and then dementia.

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Inclusions:
It is uncommon that neurons contain inclusions, the old neurons contain
lipofuscin (aging pigment) and melanin pigments in the neurons of substantia
nigra.
Dendrites:
The dendrites are cell processes arising from the soma or neuron cell body.
Most neurons have large number of dendrites to increase their surface area to
receive impulses from other neurons to transmit to nerve cell body.
Dendrites are short; they branch and subdivide repeatedly into smaller and
finer branches like a tree (arborizations). They receive large number of axon
terminals from other neurons. The cytoplasm in the dendrites is similar to
that of cell body, but devoid of Golgi complex.
Dendrite surfaces are covered with large number of small projections called
spines or gemmules which serve as sites for synaptic contacts.
Axons:
The axon is a cylindrical process; it is also called axis cylinder. The axon
diameter and length vary from one neuron to another but it retains its
diameter along most of its length.
Larger neurons have thicker and longer axons.
Axons arise from a short conical shaped region called axon hillock which
arises from the perikaryon. The axon hillock and axon do not contain rough
endoplasmic reticulum and ribosomes.

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The plasma membrane of the axon is called axolemma and its cytoplasm is
called axoplasm. In myelinated axons, the segment lying between the axon
hillock and the start of myelination is called the initial segment.
The axons give collateral branches soon after leaving the cell body. The
collaterals return to cell body. The axon distal end usually gives fine
branches called axonal terminal arborizations which end by forming
synapses.
Functions:
 Axons convey impulses away from the nerve cell body (efferent).
 Anterograde flow: transport of macromolecules along the axon from
the cell body to its terminals, promoted by a microtubule-activated
ATPase kinesin.
 Retrograde flow: axonal transport of materials taken up by
endocytosis from terminals to the cell body, promoted by the motor
protein dynein.

Synapses
A synapse is the site of contact (intercommunication) occurring between
neurons or between adjacent neurons and other effector cells like muscle or
glandular cells.
Types of synapses:
1. Axo-dendritic: between an axon and dendrites
2. Axo-somatic: between the axon and the cell body

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 3. Axoaxonic: between 2 axons
4. Dendro-dendretic between dendrites
Axodendritic and axosomatic are common in CNS.
In PNS, synapses are also present between neurons and muscles, and
glandular cells.

The structure (anatomy) of synapses:

Each synapse is composed of: terminal boutons, presynaptic membrane, and
synaptic cleft and postsynaptic membrane.
1. Bouton terminaux (terminal boutons): at the synapse, the axon terminates
in the form of bulb expansions, basket-like or club-shaped terminals. In case
the axon makes contacts along its passage, an enlargement out of the synapse
protrudes called boutons en passage. The terminal boutons contain
mitochondria, neurotubules, neurofibrilas and synaptic vesicles. The synaptic
vesicles contain chemical substances, responsible for impulse transmission
across the synapse, called neurotransmitters. They bind to specific
receptors on the postsynaptic membrane. The main types of
neurotransmitters are:
 Acetyle choline
 Catecholamines (epinephrine and norepinephrine)
 Dopamine
 Neuroactive peptides like substance P, enkephalin and endorphin.

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2. Presynaptic membrane: It is the part of cell membrane covering the end
bulb at the synapse. It appears electron dense and thicker than the rest of the
cell membrane due to accumulation of cytoplasmic proteins and filaments.
On stimulation, synaptic vesicles fuse with presynaptic membrane releasing
the neurotransmitter into the synaptic cleft.
3. Synaptic cleft: is a slender extra-cellular space, 20 - 30 nm wide, between
the pre-synaptic and post-synaptic membranes, contains hydrolytic and
oxidative enzymes.
4. Post-synaptic membrane: It is the membrane of the target neuron; it
appears denser and thicker than the rest of cell membrane. Sometimes pre-
and postsynaptic membranes are bound by bridges.
Types and functions of synapses:
1. Chemical synapses which use chemical messengers,
neurotransmitters, most common.
2. Electrical synapses which conduct neuronal signals directly, by
transmitting ionic signals through gap junctions in pre- and
postsynaptic membranes.
The arrival of an impulse at an excitatory synapse locally depolarizes the
postsynaptic membrane, whereas arriving of an impulse at an inhibitory
synapse causes local hyperpolarization with no transmission of nerve
impulse.
Excitability: change in membrane permeability in response to stimuli
Depolarization: revrsal of ionic gradient across the membrane
Action potential: wave of depolarization spread along plasma membrane
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Repolarization: plasma membrane re-establishes resting potential
Saltatory conduction: travel of action potential by jumping (faster)
Myelin Sheath:
It is a highly refractile sheath, consists of several layers of modified
plasmalemma, with higher proportion of lipids (cholesterol, phospholipids
and glycolipids). In myelinated peripheral nerve fibers, the cell membrane or
plasmalemma of the covering Schwann cells winds around the axons. The
layers of Schwann cell membrane unite to form myelin.
Formation of myelin sheath:
 The axon initially lies in a deep recess in the surface of Schwann cells
 The axon becomes enclosed by cytoplasmic process of Schwann cell
membrane, whose ends come into contact at the mesaxon
 Further development of myelin sheath will result in lengthening of
mesaxon and wrapping around the axon in a spiral manner.
 The cytoplasm between the successive turns is excluded completely,
and the inner layers of plasmalemma come into contact and finally
fuse.
 In cross section in EM, the fused leaflets of plasmalemma form the
major dense lines alternating with thin intraperiod lines.
 Mesaxon is the zone of apposition of plasmalemma of Schwann cells
The myelin sheath is interrupted at intervals by short gaps called nodes of
Ranvier. The space between two nodes is called an internode; it represents
the length of one Schwann cell.

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At the node, Schwann cells send small processes which interdigitate but do
not seal the axolemma completely. In longitudinal sections, myelin sheath
shows oblique pale areas called Schmidt-Lantermann clefts. These
represent small amount of cytoplasm trapped between the two layers of major
dense line, and may provide a path for metabolites.
In CNS, myelin sheath is produced by oligodendrocytes. Each
oligodendrocyte encloses several axons.
In H&E stains, myelin sheath appears as an empty space surrounding the
axon, since it dissolves during processing with lipid solvents. It is
demonstrated very well with osmic acid stain.

Unmyelinated nerve fibers:

CNS is very rich in unmyelinated nerve fibers which are not ensheathed
(naked).
In PNS, the unmylinated nerve fibers neurolemmal sheath by Schwann cells.
Each Schwann cell (neurolemmcyte) encloses many unmyelinated axons
without wrapping around them. Unmyelinated nerve fibers do not have
nodes of Ranvier.

Peripheral Nerve Fibers (Nerve trunk):

A nerve trunk is composed of bundles of nerve fibers. The entire nerve
(nerve trunk) is covered by tough fibrous connective tissue called
epineurium, which fills the spaces and sends extensions between the

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fascicles (interfasicular connective tissue). Each bundle is surrounded by the
perineurium.
Endoneurium is a thin layer of reticular fibers enclosing the ensheathed
axons.
The nerves function as pathways of communication between centers in CNS
and the rest of the body. The nerves that carry information from the body to
CNS is called afferent (sensory), and nerves which carry impulses from
CNS to tissues and organs of the body are called efferent (motor).
The peripheral nerves usually are mixed containing both motor and sensory.

Injuries to peripheral nerves:

Peripheral nerves might be crushed, compressed or transected, degenerative
changes followed by regenerative process take place
The nerve portion distal to the transection or crushing undergoes Wallerian or
anterograde degeneration. The proximal portion remaining attached to the
cell body undergoes regeneration. However the proximal portion shows
initially a process called retrograde degeneration.
In retrograde degeneration, the following events occur:
 Chromatolysis (dissolution of Nissl substance)
 Swelling of the perikaryon
 Movement of the cell nucleus to a peripheral position.
 Transneuronal degeneration occurs in the neurons functionally
connected to the dead neurons.

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  The axon of the proximal stump of the injured axon starts growth into
tubes of Schwann cells distal to the injury site.
 Regeneration occurs at a rate of 1 - 2 mm per day.

GANGLIA
Ganglia are aggregates of nerve cell bodies with supporting cells and
connective tissue outside the CNS.
Types of ganglia:
 Sensory (spinal)
 Autonomic (sympathetic and parasympathetic)
Spinal (dorsal root) ganglia:
These ganglia are made up of general sensory neurons and their supporting
connective tissue.
The neurons are pseudo-uni-polar with rounded or oval cell bodies of various
sizes and a single process, an axon, which bifurcates into two processes
shortly after leaving the cell body. The axons become nerve fibers either
myelinated or non-myelinated. The peripheral process joins the spinal nerve
and terminates on a sensory end organ or as a naked nerve ending. These
peripheral processes are somatic and visceral afferent fibers. The central
process joins the dorsal root and enters the cord to synapse in a specific
region.
By LM examination:

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  The ganglion appears ovoid and covered by epineurium forming a
capsule over the ganglion and blends with epineurium and
perineurium of the nerve leaving the ganglion.
 The neurons are rounded in shape with central, rounded, large and pale
staining nucleus with prominent nucleolus. The cytoplasm is rich with
fine Nissl granules.
 Each nerve cell body is surrounded by small supporting cells, called
satellite cells.
 The neurons are arranged in rows or groups at the periphery of the
ganglion, with myelinated nerve fibers running in between the cell
groups.

Autonomic ganglia:
1. Sympathetic ganglia: These ganglia are found in the sympathetic chain
alongside the vertebral column. They are made of post-ganglionic visceral
efferent neurons. Axons of neurons in the lateral sympathetic nucleus of
thoracic cord had come to the sympathetic chain via the white rami
communicantes, and many of them synapse on these ganglion cells.
In LM examination:
 The ganglion is covered with ill-defined connective tissue capsule, the
epineurium.
 The ganglion cells are more loosely arranged than in spinal ganglion.

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  The neurons are all multipolar neurons and they are more or less
uniform in size.
 The nuclei of the nerve cell bodies may be eccentrically located.
 Occasionally, binucleated, and few of them show yellow pigments.
 The satellite cells form an incomplete layer around the neurons.
 The nerve fibers in the stroma are mostly unmyelinated.

2. Parasympathetic ganglia: these ganglia lie in the walls of the organs.

Pre-ganglionic parasympathetic fibers arise from the cell bodies of the brain
stem or sacral spinal cord and go to the organs to be innervated and synapse
on the small multipolar neurons in these ganglia (postganglionic neurons).
Post-ganglionic fibers (axons) of these post-ganglionic neurons are then
distributed to smooth muscles and glands.

Glial cells (neuroglia)

Neuroglial cells are special types of cells associated with neurons. Generally,
neuroglial cells have supporting function, form myelin and have trophic and
phagocytic functions
In peripheral nervous system, the glial cells are:
 Schwann cells
 Satellite cells
In the CNS, the glial cells are:
 Astrocytes

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  Oligodendrocytes
 Microglia
 Ependymal cells
1. Astrocytes are star-shaped with numerous processes lying in cerebral
cortex. They contain numerous processes which expand over the cerebral
blood vessels giving the perivascular end feet. There are two types of
astrocytes, protoplasmic and fibrous; both can be demonstrated with silver
stain.
 The protoplasmic astrocytes possess abundant granular cytoplasm
and few microfilaments. They are found in large numbers in grey
matter; they have numerous thick cytoplasmic processes
 The fibrous astrocytes are more numerous in the white matter. They
have long slender cell processes and many microfilaments.
Both astrocytes are in direct contact with each other through the gap junctios.
The main functions of astrocytes:
 Repair and formation of scar tissue to surround the destroyed tissues.
 Structural support
 Share in blood-brain barrier (perivascular feet)
 Metabolic exchange
 Regulation of extracellular environment by absorption the excess of
neurotransmitters and releasing of enkephalins and somatostatin.
2. Oligodendrocytes are small cells with few slender processes, and small,
darkly stained nuclei. They are found in grey matter, close to perikaryons of

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pyramidal cells of cerebral cortex, as well as in white matter, where they can
be seen in rows among the myelinated nerve fibers. Their function is to
produce myelin sheath in CNS and electric insulation.
3. Microglia are small, oval cells with elongated darkly stained nuclei and
numerous short processes. They are the only CNS cells of mesodermal
origin. Their main function is phagocytosis.
4. Ependymal cells: These cells line the cavities of the brain and spinal cord
and cover the choroids plexuses. They are cuboidal to columnar in shape;
most of the cells have motile cilia on the free surfaces to create movement of
the CSF. The bases of the cells are either flat or having long processes
(tanycytes) that extend deep into the neural tissue. Tanycytes are present
mainly in the floor of third ventricle.
5. Schwann cells (neurolemmocyte): these are the supporting cells of the
PNS. They form the neurolemmal sheath of unmyelinated nerve fibers, or the
myelin sheath in myelinated nerve fibers. Their function is to produce
myelin sheath and electric insulation in PNS.
6. Satellite cells are found around the nerve cell bodies of the ganglia of PNS
such as spinal or sympathetic ganglia.

The Central Nervous System

The central nervous system is composed of gray and white matters. The gray
matter contains the cell bodies of neuron, neuroglia, capillaries and
unmyelinated nerve fibers. The grey matter is present in the central H-shaped
area of spinal cord and cerebral and cerebellar cortices.
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The white matter of CNS contains myelinated axons arranged in tracts,
neuroglia (oligodendroglia and fibrous astrocytes), and few capillaries but no
cell bodies of neurons.
The CNS is surrounded by an outer dense fibrous dura matter, middle
arachnoid and an innermost pia matter. The arachnoid sends trabeculae
connecting with pia mater enclosing the subarachnoid spaces filled with
cerebrospinal fluid (CSF). In some locations, the arachnoid perforates the
dura forming protrusion which terminate in venous sinuses of the dura and
covered by endothelial cells of the veins. These protrusions are called
arachnoid villi. The CSF filling the CNS cavities is produced by the choroids
plexus.
Spinal Cord
The spinal cord is composed of an H-shaped central core of gray matter
surrounded by an outer layer of white matter. The proportion of gray to
white matter differs according to the level of the cord.
The gray matter is composed of two large lateral masses connected by the
gray commissure, in which lies the central canal which is largely obliterated
in adults. Each lateral gray mass has a dorsal gray horn or column, a ventral
gray horn or column and an intermediate gray area. The shape of gray matter
varies in different parts; the ventral horns are widest in the cervical and
lumbar enlargements in which the multipolar anterior horn cells innervating
the upper and lower limbs are located. The ventral horns are narrowest in
thoracic segments. The gray matter contains:
 neuron cell bodies together with their dendrites
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  the initial segments of the neurons axons
 unmyelinated nerve fibers
 the termination of myelinated nerve fiber tracts
 numerous blood vessels
The white matter is composed of:
 nerve fibers (mostly myelinated)
 neuroglia
 connective tissue septa from pia matter
 blood vessels coming in connective tissue septa
Dorsal roots (incoming sensory fibers) are attached to the dorsolateral surface
of the cord (dorsal sulcus), while the ventral roots are attached to the
ventrolateral surface (ventrolateral sulcus). In relation to these sulci, the
white matter is divided into dorsal white columns, lateral white columns, and
ventral white columns.
The spinal cord is covered by meninges; dura matter is the outermost,
followed by the arachnoid then the pia matter which is the innermost.
Pia matter is a fibrous membrane attached to spinal cord, from which
connective tissue septa carrying blood vessels penetrate the white and gray
matter.
When the dorsal root fibers enter the cord and the ventral root fibers leave the
cord, they lie in the subarachnoid space.

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When the root fibers exit through the intervertebral foramina, the arachnoid
matter disappears quickly, while the dura matter continues over them as
epineurium.
The dorsal gray column contains a large pale area (substantia gelatinosa), and
the entrance of incoming sensory fibers into the dorsal gray column.
Thoracic segments contain lateral extension of the intermediate gray
containing small multipolar neurons (sympathetic nuclei).

Cerebellum
The cerebellum is a highly folded structure with pia matter extending down
between the folds. Each fold has a central core of white matter made of nerve
fibers surrounded by a cortex of grey matter which is made of three layers:
1. Outer molecular layer, which is largely made of nerve fibers
with scattered small neuron cell bodies
2. Middle or Purkinje cell layer.
The Purkinje cells are multipolar neurons which have flask-shaped cell
bodies and highly developed dendrites which occupy most of the
molecular layer.
All the incoming fibers to cerebellar cortex synapse indirectly on
Purkinje cells.
The axons of Purkinje cells extend down through the granular layer
into the white matter and terminate in the central cerebellar nuclei.
These are the efferent fibers of the cerebellum.

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 3. Inner granular layer which is composed of small, densely
packed, atypical neuron cell bodies, with darkly-stained nuclei
and very little cytoplasm.

Cerebrum
The gray matter of the cerebral cortex consists of a number of layers of
neuron cell bodies of different shapes and sizes. These layers are named
according to the appearance of the cells. Most of these cells are association
cells, they receive fibers from various parts of the cortex and their axons pass
to other parts of the cortex.
Axons of the large pyramidal cells of the motor cortex (upper motor neurons)
terminate indirectly on the motor neurons of the brain stem and on the
anterior horn cells in the spinal cord.
The surface of cerebral cortex is covered with pia-arachnoid.
The cerebral cortex is made of the following layers from superficial to deep:
1. Molecular layer is made of small horizontal cells of Cajal and the
dendrites of neurons in the deeper layers.
2. Outer (external) granular made of small pyramidal cells. They have short
axons that pass to the deep cortical layers.
3. Outer (external) pyramidal layer made of medium-sized pyramidal cells
4. Inner granular layer made of stellate cells.
5. Inner pyramidal layer is made of large pyramidal cells of Betz in motor
cortex. The apices of pyramidal cells are directed towards the surface. They
have basal and apical dendrites, with spines, which spread laterally.
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6. Polymorph (multiform) layer made of irregularly shaped neurons. Most
of cells are fusiform that have basal short dendrites and long apical dendrites
directed to the surface.

Choroid Plexus
The central canal and the ventricles of the CNS are lined with ependymal
cells. These cells are covered externally with pia matter and constitute the
vascular tela choroidae. From the vascular tela choroidae, complex
evaginations project into the lumen of the ventricles forming the choroid
plexus.
The core of the evagination is highly vascularized connective tissue
containing many capillaries and small blood vessels. The covering
epithelium is modified ependymal cells, cuboidal or low columnar cells with
very granular cytoplasm.
The cerebrospinal fluid (CSF) is secreted by choroids plexus and ventricular
ependyma. CSF is a colorless, clear fluid containing small amount of protein,
few cells, glucose and sodium and potassium chloride. CSF circulates
through the brain ventricular system to subarachnoid space. Impairment of
CSF normal flow leads to its accumulation and hydrocephalus.

Blood-Brain Barrier:
It is a functional barrier which restricts the passage of circulating substances
from the blood to the brain for protection. However, blood-brain barrier is
lacking in some areas of the brain (choroid plexus, pituitary and pineal glands
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and vomiting center of hypothalamus). The blood-brain barrier is composed
of:
 Occluding tight junctions between the endothelial cells of blood
capillaries, the main component
 Continuous basal lamina of capillary endothelium within the brain
(non-fenestrated) and luminal neuroactive humoral substances which
destroys neurotoxic metabolites.
 Few or no pinocytotic vesicles to mininmize transcapillary transport.
 Perivascular end-feet of astrocytes

Nerve endings or nerve terminals

Nerve endings are classified into afferent (sensory) or efferent (effector)
types.
I. The sensory nerve endings:
Functionally the sensory nerve endings might be classified into special (taste,
vision and olfaction), and general. The general type is classified into
exteroceptors (in skin), interoceptors (in viscera) and proprioceptors (in
muscles, joints and tendons).
Structurally, sensory nerve endings may be classified into non-encapsulated
and encapsulated.
A. Non-encapsulated or unencapsulated nerve endings or free nerve
endings which consists of the terminal branches of sensory nerve fibers lying

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freely in skin and corneal epithelia, as well as dermis, fascia, tendons,
ligaments and muscles.
Myelination terminates prior to nerve endings and the axon:
 May terminate in the squamous epithelium of skin (pain fibers)
 May terminate as cup-like endings in contact with Merkel`s cells
(Merkel`s tactile disc), for touch.
 May coil around the hair follicles forming peritrichial plexus, for
touch.

B. Encapsulated nerve endings: where the nerve endings is covered by a

thin or thick capsule.
1. Meissner`s corpuscles are best seen within the dermal papillae of thick
skin. They are composed of an inner core of Schwann cells and nerve
terminals surrounded by thin collagen bundles and fibroblasts. They are
receptors for discriminatory tactile touch.
2. End bulb of Krause is spherical corpuscle, surrounded by a thin capsule,
larger than Meissner`s, and present in the connective tissue of mucous
membranes. They are stimulated by cold temperatures.
3. Ruffini`s corpuscles are spindle-shaped, encapsulated nerve endings
found in joints and dermis. They are mechanoreceptors responding to joint
movements.

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4. Pacinian corpuscles are formed of small core where the terminal axon is
located. The core is surrounded by concentric layers of fibroblasts alternating
with thin collagen fibers giving the appearance of sliced onion.
Function: proprioception for deep pressure and vibration sense.
5. Muscle spindles: Skeletal muscle is supplied by afferent (sensory) nerve
endings sensitive to stretch located in tendons or muscle fibers
(proprioceptive mechanoreceptors). They are present within groups of
specialized, spindle-shaped muscle fibers or tendons called muscle spindles
or tendon spindles. These muscle spindles consist of modified striated
muscle fibers and their associated nerve fibers; they are most numerous in
muscles of extremities and least in extraocular muscles.
Muscle fibers lying inside the spindles are called intrafusal; they are smaller
than the extrafusal (ordinary) muscle fibers and innervated by gamma
efferent fibers (motor).
The muscle spindles are of two types:
a. The nuclear bag fibers containing a group of nuclei in the
expanded, non-striated mid region, with an afferent primary nerve
ending coiled around the midregion (Annulo-spiral ending).
b. The nuclear row or chain in which the nuclei are arranged in a
row or chain with a primary nerve ending (annulospiral) coiled around
the midregion plus two secondary endings, one on each side, called
flower spray endings.

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Function of muscle spindle is to detect the changes in muscle tension and to
convey the information to CNS (spinal cord); important in maintaining the
posture and regulation of the voluntary movements.

Taste buds:
Taste buds are present in circumvallate, foliate (rabbit), and fungiform
papillae. The taste bud looks barrel-shaped, consists mainly of three types of
cells.
 Supporting (sustentacular) cells: slender, spindle-shaped cells, thin
darkly-stained ovoid nuclei; they also have microvilli; they are
peripherally located.
 Gustatory or taste receptor cells: more numerous light-staining, round
cells (neuroepithelial cells) with euchromatic nuclei. These cells have
microvilli and their bases are surrounded by fine nerve fibers carrying
taste sensation. The taste pore opens in the trenches around the
circumvallate papillae.
 Basal cells: they are short cells, can differentiate to other types of
cells.
 By EM, taste buds are composed of four types of cells, type I, type II,
type III and the fourth type which is made of undifferentiated basal
cells.

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The olfactory mucosa:
The olfactory mucosa present at the roof of nasal cavity, and the upper part of
the nasal septum. It is made of pseudostratified columnar ciliated epithelium;
when examined by LM three types of cells can be distinguished:
1. Olfactory cells: they are bipolar neurons which contain the smell receptors.
They have non-motile cilia which posses the receptors for smell.
2. The supporting cells are tall with apical nuclei.
3. The basal cells are short and round located at the basal lamina of the
epithelium.

II. The efferent terminals:

The motor end-plate (myoneural junction):
On the striated muscle fibers near the motor end plate, the myelinated axon
loses its myelin sheath and branches to terminate on the muscle fibers
forming a dilated termination (terminal bouton). The terminal axon is
covered by Schwann cell cytoplasm. The terminal part of the axon contains
numerous mitochondria and synaptic vesicles containing neurotransmitter,
acetylcholine. Between the axon terminals and the muscle cell membrane is
a space called synaptic cleft containing the matrix of the basal lamina. At the
junction, the axolemma is called the presynaptic membrane and the
sarcolemma is called the postsynaptic membrane.

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 Circulatory System
The circulatory system consists of cardiovascular and lymphatic vascular
systems.
The cardiovascular system consists of the heart, arteries, capillaries and
veins.
The lymphatic vascular system consists of lymphatic capillaries and
lymphatic ducts of different sizes.
The circulatory system is divided into two components:
a. Macrovasculature includes vessels with diameter more than 0.1
mm (large arterioles, muscular arteries and veins and elastic
arteries.
b. Microvasculature includes vessels with diameter less than 0.1
mm such as arterioles, capillaries and postcapillary venules.
Microvasculature is important in interchange of gas and
nutrients between tissue and blood.
Basically, the walls of blood vessels (except the smallest ones) are composed
of three coats or tunics with a specific thickness for a particular size vessel,
with variable amount of connective tissue and muscle according to the size
and function of the vessel. The three coats are:
 Tunica intima: endothelium with basement membrane and delicate
collagen fibers
 Tuniaca media: smooth muscule fibers, elastic fibers, may contain
vasa vasora extending from adventitia. The media has the main effect

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 on blood flow. The amount of elastic fibers decreases and that of
smooth muscle increases with decreasing diameter of arteries.
 Tunica adventitia: it is a supporting layer, rich in collagen fibers to
prevent over-extension and permit some flexibility; it contains vasa
vasorum
For teaching purposes, blood vessels are classified according to their
diameters into small, medium-sized and large.

Vasa vasora:
Vasa vasora means vessels of the vessel. Vasa vasora are small
vessels present in the adventitia and the outer part of the media of
large blood vessels. They supply oxygen and metabolites to the outer
layers of large blood vessels (media and adventitia) which cannot be
nourished by diffusion from the circulating blood.

Nerve supply of blood vessels (innervations):

Blood vessels are richly supplied by networks of unmyelinated sympathetic
(adrenergic vasomotor) nerve fibers causing vessel constriction. In skeletal
muscles, arteries receive an additional cholinergic vasodilator nerve supply.

Blood Capillaries
Blood capillaries are thin-walled tubes whose walls consist of rolled up
endothelial cells and intermittent strands of connective tissue referred to as

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adventitia. Their length may reach up to 50 µm, and their diameter might be
7 – 9 µm; slightly wider than that of the red blood cell.
Structure:
The capillary walls consist of a single layer of endothelial cells whose
external surfaces rest on basal lamina and enclose pericytes; the basal lamina
produced by the endothelial cells. The advetitia is made of intermittent
strands of connective tissue. No media.
The endothelial cells are elongated with large nucleus bulging into capillary
lumen. Their cytoplasm contains small Golgi complex, ribosomes,
mitochondria, some RER, intermediate filaments and microfilaments. Zonula
occludens are present between endothelial cells.
Pericytes are undiffrentiated, of mesenchymal origin, pale staining cells with
nuclei bulging outwards, associated with the endothelium of blood capillaries
and postcapillary venules. Pericytes are enclosed within their own basal
lamina which fuses with endothelial cells basal lamina. Pericytes are
contractile since they have myosin, actin and tropomyosin filaments in their
cytoplasm, and their slender cytoplasmic processes wrap around the
associated capillary to assist constricting its lumen.

Functions of blood capillaries:

 Gas exchange between blood and tissues
 Metabolic exchange between blood and the surrounding tissues.
 Transport of hormones from cells to target tissues

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  Anti-thrombogenic effect by preventing the contact between
thrombocytes and subendothelial connective tissue on endothelial cells
desquamation.
Besides these functions, there are other functions performed by endothelial
cells such as:
1. Convert angiotensin I to angiotensin II.
2. Inactivate certain biological compounds such as bradykinin, serotonin,
norepinephrine, thrombin and prostaglandins.
3. Lipolysis of lipoproteins into triglycerides and cholesterol.
4. Production of vasoactive factors such as endothelins (vasoconstrictor) and
nitric oxide (vasorelaxant).
5. Produce vascular endothelial growth factors important for vessels
formation during embryonic life and well being of vessels in adulthood.

Types of capillaries:
 Continuous capillaries (somatic): the endothelial cells rest on
uninterrupted well-developed basal lamina, with no fenestration and
their edges are held together by tight junctions. Endothelial cells
(except in nervous system) show numerous pinocytotic vesicles on
both surfaces and in the cytoplasm sometimes fuse to form
transendothelial channels for macromolecule transport. This type of
capillaries is most numerous, present in muscles, connective tissue,
exocrine glands and nervous system (blood-brain barrier).

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  Fenestrated capillaries (visceral): the wall of these capillaries
exhibits fenestrations (60 -80 nm in diameter) in the endothelia closed
by thin diaphragms (except in renal glomeruli). These diaphragms are
thinner than the cell membrane and do not have the trilaminar
structure. The endothelial basal lamina is continuous. This type of
capillaries is present in kidney, small intestine and endocrine glands.
Pericytes are rare.
 Discontinuous sinusoidal capillaries: the walls of these capillaries
are discontinuous with multiple fenestrations in endothelial cells with
no diaphragms. The endothelial cells are widely separated from each
other and the basal lamina is discontinuous. Capillary diameters are
greatly enlarged 30 – 40 um, to help slowing blood circulation.
Macrophages are present among the endothelial cells.
This type of capillaries is present in liver, bone marrow and spleen.
The main function of sinusoidal capillaries is to enhance the exchange
between blood and tissues.

Arteriovenous Anastomoses
Arteriovenous anastomoses (A-V shunts) are direct connections between
arterial and venous systems that divert blood from the capillaries. The
contraction of smooth muscle in the arterioles of A-V shunt directs the blood
to capillary bed, and relaxation sends blood to venules bypassing capillaries.
(Some authors say that capillary sphincter conctraction directs blood through
the shunt). The A-V shunts are present in skin of fingertips, nose, and lips
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and erectile tissue of penis and clitoris. In ears, fingerpads and fingernail
beds and toes, the arterial side of the anastomosis forms a special organ called
glomus or glomera (plural), where the arteriole coils into a ball-like structure
and its media becomes very thick.
Function: thermoregulation, and regulation of the blood flow and blood
pressue.
Metarterioles
Metarterioles arise from arterioles just before giving origin to capillary beds,
where a discontinuous layer of smooth muscles surrounds the endothelial
lining. However, just before the beginning of capillary bed at the arteriolar-
capillar junction, metarterioles exhibits a circular layer of smooth muscle,
called precapillary sphincter which controls the blood flow within the
capillary beds.
Arterioles
Arterioles are the terminal branches of arterial tree; they impart peripheral
resistance that regulates the blood pressure and the blood flow to capillary
beds. Arterioles diameter is generally less than 0.5 mm (40 to 400 um).
Their walls are composed of:
 Tunica intima is very thin, made of endothelium and very thin layer
of subendothelial reticular fibers. The internal elastic membrane is
absent in the small arterioles, but starts to appear as thin intermittent
in the large arterioles.

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  Tunica media is made of one or two layers of smooth muscle fibers
circularly arranged (may reach six layers in large arterioles). No
external elastic membrane.
 Tunica adventitia is thin, contains collagen fibers and blends in with
the surrounding connective tissue.

The Weibel-Palade granules:

The Weible-Palade granules are rod-shaped granules found in the endothelial
cells of vessels larger than capillaries. These granules contain a protein of the
blood coagulation mechanism, factor VIII, also called Von Willebrand factor.
Deficiency of this factor leads to prolonged bleeding time and haemophilia.

Medium-sized arteries
Medium-sized arteries are also called muscular arteries because their media is
formed mainly of smooth muscle cells. Most of the named distributing
arteries are of medium-sized type such as: radial, femoral, cerebral and
coronary arteries. Their function, denoted by their names, is to distribute
blood to various parts of the body.
Their wall is composed of three tunics:
1. Tunica intima is composed of:
 A single layer of simple squamous epithelium called
endothelium.
 A very thin layer of subendothelial connective tissue.

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  Prominent internal elastic membrane (lamina)
separates tunica intima from tunica media.

2. Tunica media is made of compact circumferentially arranged smooth

muscle cells (up to 40 layers) intermingled with elastic fibers, reticular fibers
and proteoglycans, all secreted by smooth muscle fibers. Prominent external
elastic membrane or lamina is present between the media and advetitia.

3. Tunica adventitia is thinner than the media; it consists of collagen and

elastic fibers, few fibroblasts, lymphatics, nerves and vasa vasora (in large
muscular arteries).
Some arteries are modified according to location and function such as:
cerebral arteries have thin wall, coronary arteries have thick walls and large
amount of elastic tissue, reduced muscle and elastic fibers in pulmonary
arteries, very thick media in umbilical arteries.

Large artery (elastic)

The large elastic arteries include the aorta arising from left ventricle, the
pulmonary trunk arising from the right ventricle and their major branches.
The high content of elastic lamellae (elastin) in their media gives them the
yellowish color in fresh state and is responsible for the phenomenon of elastic
recoil of these arteries which is important for maintenance of the arterial
blood pressure during diastole. The wall has 3 coats:
1. Tunica intima consists of:
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  Endothelium of simple squamous type containing rod-shaped
cytoplasmic inclusions called Weibel-Palade bodies containing von
Willebrand factor (coagulating factor VIII).
 Thick layer of subendothelial connective tissue containing
longitudinally oriented collagen fibers, fibroblasts and smooth muscle
cells; both cells secrete extracellular matrix.
 Internal elastic membrane is present but hard to distinguish from the
media elastic lamellae.
2. Tunica media is very thick, comprises almost the whole wall; it is
composed of large number (40 in newborns and 70 in adults) of fenestrated
concentric elastic lamellae with relatively few smooth muscle fibers, reticular
fibers in between, proteoglycans and glycoproteins. The external elastic
membrane is ill-defined. The outer half of adventitia is penetrated by vasa
vsora.
3. Tunica adventitia is underdeveloped, contains collagen and elastic fibers,
nerves, lymphatics and small blood vessels for nourishment called vasa
vasorum which send capillary branches to the media.

Venules
Venules are thin-walled vessels slightly larger than capillaries; they drain the
capillary networks. Transition of capillaries to venules is gradual and gives
rise to postcapillary or pericytic venules with a diameter of 0.1 to 0.5 mm.

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Their intima consists of loosely arranged endothelium and the media contains
only pericytes occasionally enclosed within the basal lamina.
The media of large venules contains few smooth muscle fibers, but small
venules do not have media.
The adventitia is thin composed of collagen and few elastic fibers.
Their function, resembles those of blood capillaries, is to exchange
metabolites between blood and tissues and to produce local swelling in acute
inflammation.
Medium-sized (muscular) veins
Medium-sized veins are thin-walled vessels with diameter of 1 – 9 mm. The
wall is made of:
 Tunical intima is made of endothelium and thin subendotleial layer
which is absent sometimes. Poorly developed or absent elastica
interna (internal elastic membrane).
 Tunica media consists of small bundles of smooth muscle fibers with
some reticular and elastic fibers. It is much thinner and less compact
than an artery of the same size.
 Tunica adventitia is well developed, thick and made of longitudinally
oriented collagen and elastic fibers, and fibroblasts.
Differences between the medium-sized arteries and veins:
1. Arteries have patent rounded lumen while the veins usually have wider
lumen which tends to be collapsed.

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2. The three coats are well distinct in arteries, and less distinct in veins.
Arteries have much thicker and compact tunica media compared to that of the
veins. The media of the veins is thinner and the muscle fibers are loosely
arranged.
3. The veins lack both the internal and external elastic membranes which are
well developed in the arteries.
4. The tunica adventitia of the veins is thicker than its media, compared to
that of the arteries.
5. The walls of the veins, in general, have more connective tissue and more
loosely arranged than those of the arteries.
6. Veins have valves and arteries do not.

Large veins
The large veins include vena cava (superior and inferior), azygos, splenic,
portal and external iliacs. The wall structure is:
1. The tunica intima is well developed and made up of endothelium and
subendothelial connective tissue.
2. The tunica media is very thin and made up of few layers of smooth muscle
cells and abundant connective tissue.
3. Tunica adventitia is a wide area and well developed. It contains collagen
and elastic fibers and longitudinal bundles of smooth muscles. The adventitia
of vena cava and pulmonary veins contains some cardiac muscle fibers for a
short segment before entering the heart. Vasa vasorum are also present.

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Large and medium-sized muscular veins have semilunar valves at their
interior to prevent the retrograde flow of blood. These valves consist of
elastic tissue and are lined with endothelium on both sides and project into
the vein lumen.
The Heart
The heart is a muscular organ which pumps blood into the circulatory system
by rhythmic contractions. The heart walls consist of three tunics (coats), an
inner endocardium, middle myocardium and an outer epicardium.
The fibrous skeleton of the heart lies in the central region and serves as the
base for valves and the site of attachment for cardiac muscle fibers.
1. The endocardium is equivalent to tunica intima; it lines the heart
chambers and valves. It is composed of:
 Endothelium: single layer of simple squamous epithelium
 Subendothelial layer of loose connective tissue containing elastic and
collagen fibers as well as smooth muscle fibers.
 Subendocardial supporting layer: contains dense connective tissue and
merges with the connective tissue surrounding the bundles of cardiac
muscle fibers (perimysium). It contains the branches of the heart
impulse conducting system, veins and nerves.
2. The myocardium is the thickest, comprising the whole mass of the heart;
it corresponds to tunica media in blood vessels. It is composed of cardiac
muscle cells arranged in a complex spiral manner. Loose connective tissue
invests the muscle fibers (endomysium) rich in blood capillaries and

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lymphatics. The cardiac muscle cells are joined end to end by intercalated
discs, desmosomes at the transverse part of the disc and gap junctions at the
longitudinal part. The desmosomes prevent separation of cells during
contraction and the gap junctions help to spread the wave of excitation along
the entire myocardium.
3. The epicardium is the visceral layer of pericardium, a serous coat covered
externally by simple squamous epithelium (mesothelium). It corresponds to
tunica adventitia of blood vessels; it is composed of:
 Inner layer of fibroelastic connective tissue containing blood vessels
of the heart (coronaries), nerves, ganglia, lymphatics and adipose
tissue. This layer blends with the connective tissue covering the
bundles of heart muscle.
 Outer or superficial mesothelial layer made of simple squamous
epithelium and thin submesothelial layer of connective tissue.

The Fibrous Skeleton of the Heart

The heart skeleton is composed of dense connective tissue (in some areas
fibrocartilage) arranged into annuli fibrosi, trigona fibrosa and septum
membranaceum.
The annuli fibrosi are rings of dense connective tissue around the
atrioventricular orifices and those of the origin of aorta and pulmonary artery.
The rings around the aortic and pulmonary artery origin give attachment to
atrial cardiac muscles; the rings around the A-V orifices give attachment to
the ventricular cardiac muscles.
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The septum membranaceum lies at the upper part of interventricular septum.

The Heart Valves

The heart valves are located at the atrioventricular (A-V) orifices and those
between the heart great vessels (tricuspid, pulmonary, aortic and mitral).
The valves are folds of endocardium, composed of two or three leaflets; each
of which consists of a central core of dense connective tissue of collagen and
elastic fibers and lined on both sides by endothelium.
Valve bases are attached to annuli fibrosi of cardiac skeleton and their
ventricular surface is connected to chordae tendineae of papillary muscles.

The Cardiac Conducting System

This system consists of modified cardiac muscle fibers to initiate impulses
and conduct them rapidly throughout the heart. It is composed of S–A node,
A–V node, A–V bundle of His and Purkinje fibers.

The Sinoatrial Node (S - A node):

S-A node is the pacemaker of the heart; it is located near the junction of
superior vena cava with the right atrium. It is composed of modified small
cardiac muscle cells with few myofibrils (nodal cells). The nodal cells are
arranged around a nodal artery.
Contraction of cardiac muscles is initiated at S-A node, spreads through the
atria, and then propagated to A-V node.
S-A node is supplied by autonomic nerve fibers.
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The atrioventricular node (A-V node):
It is located in the interatrial septum near the opening of coronary sinus. Like
the S-A node, it consists of modified cardiac muscle fibers with cytoplasmic
projections in various directions forming networks.
A-V node has nodal artery and is supplied by autonomic nerve fibers.
A-V node conducts impulses to ventricular myocardium through the A-V
bundles.

The atrioventricular bundle of His:

The A-V bundle originates from the A-V node and is composed of similar
cells; it penetrates the fibrous ring at the atrioventricular orifice and divides
into right and left bundle branches which run distally on either side of the
interventricular septum beneath the endocardium.
The two bundle branches are made of larger distinctive cardiac muscle cells
called Purkinje fibers.

Purkinje fibers:
These are modified cardiac muscle fibers specialized in impulse conduction.
They appear larger than the ventricular fibers with fewer myofibrils at the
periphery. They have central nuclei (may be binucleated) surrounded by
clear area. The cytoplasm is rich in glycogen and mitochondria.
Purkinje fibers supply the apex of the heart before its base, and hence
ventricular contraction starts at the apex then spreads towards the base.

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Innervarvation:
Heart is supplied by autonomic nervous system, both sympathetic and
parasympathetic.
Stimulation of parasympathetic division (vagus nerve) slows the reart rate,
while stimulation of sympathetic increases the heart rate.
Free nerve endings for pain sensation are present between the cardiac muscle
fibers.
Lymphatic vascular system
It is made of thin-walled channels, lined with endothelium. It collects lymph
from the tissue spaces and returns it to the circulation. Due to presence of
many valves in the lymph vessels, lymph circulates only in one direction
towards the heart by the action of smooth muscles in the vessel wall and is
aided by contraction of the surrounding skeletal muscles.

The lymphatic capillaries:

They are thin-walled, blind-ended vessels which consist of single layer of
endothelium. They have large diameter, have no fenestrations in their
endothelium, no zonula occludens between the cells and no basal lamina.
Their lumens are kept opened by numerous microfibrils.

Lymphatic ducts:
These occur by the union of small, thin lymphatic vessels. The end result is
the formation of thoracic duct and the right lymphatic duct which empty in

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the large veins. Their structure is similar to that of the veins. They have
three coats.
1. Tunica intima is made of endothelial cells and a thin layer of connective
tissue.
2. Tunica media is made up of made of bundles of smooth muscle fibers
arranged longitudinally as well as circularity.
3. Tunica adventitia is underdeveloped and is made of connective tissue with
some smooth muscle fibers, nerves and vasa vasora.

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 Immune System and Lymphoid Organs
The immune system is a system of cells which imparts immunity to the body
in order to protect it from invasion by microorganisms and foreign
substances. Its function is closely associated with lymphatic system.
The immune system is composed mainly of:
 Cells present in blood, lymph, epithelium and connective tissue (T and
B lymphocytes, macrophages, dendritic-antigen presenting cells and
eosinophils).
 Central lymphatic organs: site of proliferation and differentiation of T
and B lymphocytes
 Peripheral lymphatic organs: site of lymphocyte proliferation and
exposure to antigens
 Lymphoid nodules (MALT) present in the mucosa of digestive system,
respiratory system, reproductive and urinary system.

An antigen means any foreign substance recognized as non-self by the body

immune system; when introduced into the body, it produces a specific
immune response. The cells of immune system recognize small particles of
the antigen molecules called epitopes or antigen determinants.

Antibodies:
The substance produced specifically in response against the antigen is a
special kind of protein called antibody.

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Antibodies are made of glycoproteins and react specifically with antigen
epitopes. Antibodies could be free circulating molecules or could be integral
proteins in the membranes of lymphocytes.
An antibody molecule is a Y-shape molecule composed of two identical
heavy chains and two identical light chains connected by disulfide bonds.
The isolated shaft of the Y is composed only of heavy chains and forms the
FC segmenet of the molecule which binds to receptors present on the cell
membranes of several cells. The amino terminal ends (NH2) of the
molecules (made by light and heavy chains) are variable and compose the
antigen binding sites of the molecule.
Cells of the immune system are able to distinguish self (body own tissue)
from non-self (foreign) substances. In autoimmune disease (AIDS), the body
reacts against its own self tissue considering it as an antigen or foreign tissue.

Actions of antibodies:
 Neutralization: To precipitate the soluble antigens and neutralize
their harmful effect on the body.
 Opsonization: To enhance (stimulate) phagocytosis by opsonization
= coating the antigens or the invading micro-organisms by antibodies
produced against them. The surface of phagocytic cells contains
receptors for the Fc segment of IgG molecules. Phagocytosis is
mediated by interaction between the Fc segments of the antibody and
the Fc receptors on the membranes of phagocytic cells.

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  Activation of complement system: Some antibodies initiate cell lysis
of the invading micro-organism by acting on its cell membrane. The
surface of phagocytic cells contains receptors for the Fc segment of
C3 proteins of the complement system which opsonize pathogens to
enhance phagocytosis.

Classes or types of immunoglobulins (antibodies):

 IgG is most abundant, it forms 75% of serum immunoglobulins. It is
the only antibody that crosses the placental barrier to protect the fetus
and the newborn against infection, since it is very effective in killing
microorganisms. Macrophages, neutrophils and eosinophils have
receptors on their surfaces for its Fc segment.
 IgA present in small amount, mainly in tears, colostrum, saliva,
nasal, bronchial, intestinal and prostatic secretion and vaginal fluid as
a dimer or trimer joined by protein J. It prevents proliferation of
microorganisms and protects mucous surfaces.
 IgM (~ 10%) found as pentamer on the surface of B-lymphocytes,
the first antibody to be produced in the primary immune response. It
acts as a specific receptor for antigens and activates the complement
system.
 IgD found on the surface of B-lymphocytes, serves as receptor for
specific antigens.

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  IgE has a great affinity for receptors on plasma membranes of mast
cells and basophils; it participates in allergic reactions and killing the
parasitic infestation.

Types of immune resposes:

There are two types of immune responses, innate and adaptive.
 Innate or natural: non-specific, fast, older than the other type; it
occurs through the cells neutrophils, macrophages, mast cells, and
natural killer cells.
 Adaptive or acquired: slower than innate, specific, depens on
recognizing antigens by B and T lymphocytes and produces memory
cells. This response is more recent than the innate, and has two
subtypes:
1. Humoral immune response which results due to production
of antibodies by plasma cells derived from B lymphocytes.
This response is mediated by B lymphocytes and plasma cells
and may require macrophages and helper T- lymphocytes. The
plasma cells that develop as a result of immune response secrete
antigen-specific circulating immunoglobulins or humoral
antibodies. B lymphocytes that did not differentiate into plasma
cells remain as memory cells
2. Cellular immune response is mediated by T-lymphocytes
which:

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  Act as cytotoxic cells directly by killing the abnormal cells
(jnfected by virus or bacteria and tumor cell) which have foreign
epitopes on their surfaces.
 T lymphocytes secrete cytokines (lymphokines) which act on B
and T lymphocytes, macrophages, and neutrophils.
 T lymphocytes can only recognize antigen bound to major
histocopmpatability complex or protein (MHC). This response is
mediated by T helper and T suppressor; it creates antigen specific
killer cells. This response is responsible for transplant rejection.

Cells of the immune system:

The cells of the immune system are lymphocytes, plasma cells, mast cells,
neutrophils, eosinophils, macrophages and dendritic cells. These cells are
distributed as isolated cells throughout the different tissues, or as solitary
lymphoid nodules in different organs or present in the lymphoid organs such
as lymph nodes, spleen, bone marrow and MALT.
Lymphocytes are the only cells which are able selectively to recognize
antigens due to presence of specific receptors on their surfaces. There are
two types of lymphocytes: B and T lymphocytes. Although the short
microvilli are more numerous on B lymphocytes than on T lymphocytes,
morphologically, it is hard to differentiate between B and T cells either by
LM or EM. They can be differentiated by immunocytochemical methods due
to presence of different surface protein markers.

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1. B lymphocytes
All lymphocytes originate in the bone marrow. B-lymphocytes differentiate
in the bone marrow into immunocompetent cells (can respond to antigen),
and then migrate to secondary lymphoid organs as mature cells. B is an
abbreviation for bursa of Fabricius in birds where their B lymphocytes
mature. In humans, bone marrow is equivelant to bursa of Fabricius.
B lymphocytes constitute the minority (5-10%) of the circulating
lymphocytes. They have numerous short microvilli and surface
immunoglobulins as well as large number of receptors for specific antigens.
B lymphocytes recognize the soluble and the surface antigens.
Most circulating B lymphocytes are naive cells which have not been exposed
to antigens. In lymphoid organs, when B-lymphocytes are exposed to a
foreign antigen, they are activated to proliferate and differentiate into plasma
cells which secrete antibodies against that antigen. Activation of B
lymphocyte is assisted by T-helper lymphocytes.
Some B-lymphocytes do not differentiate into plasma cells but they stay as
long-lived B memory lymphocytes which react quickly and strongly to a
second exposure to the same antigen.

2. T-lymphocytes form the majority (65-75%) of the circulating

lymphocytes. They originate in bone marrow and migrate to thymus
gland where they mature and proliferate, and acquire the surface markers:
CD2, CD3 and TCR (T cell receptors).

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Lymphocyes then are carried to thymus-dependant zones of other lymphoid
organs by blood stream (naive lymphocytes). The T cell markers and
receptors enable T lymphocytes to recognize the epitopes, small molecular
domains of the antigen, such as those of the major histocompetability
complex (MHC).

Memory T cells persist as long-lived cells after an infection has resolved.

Memory T cells increase in number after exposure to the same antigen; They
might be either CD4 (helper T) or CD8 (cytotoxic) cell.
There are four populations of T-lymphocytes:
 Helper T cells (CD4 cells): secrete interlukins (cytokines) and
stimulate the differentiation of B-lymphocytes into plasma cells,
activate B lymphocytes to produce antibodies, regulate cytotoxic T
cell function and activate macrophages (delayed hypersensitivity). T
helper cells are killed by HIV virus.
 Regulatory T cells, formerly known as suppressor T cells: regulate
cellular and humoral immunity, and inhibit the action of helper and
cytotoxic T cells i,e they depress the antibody production by
depressing the differentiation of B lymphocytes into plasma cells.
 Cytotoxic T cells (CD 8 cells): kill malignant or virus-infected cells by
two ways: 1. produce the protein perforin which causes holes in cell
membrane then lysis. 2. They activate the genes responsible for
programmed cell death (apoptosis). The cytotoxic T cells are

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 responsible for tissue grafts and transplant rejection and cell-mediated
immunity.
 Gamma/delta (γδ) T lymphocytes: these are small group of
lymphocytes (5% of circulating T cells) that lack CD4 and CD8.
They develop in thymus gland then migrate to epithelial tissue of the
intestine, skin or vagina. They function as the first line of defence
against the invading organisms.
3. Natural killer cells (NK):
Unlike B and T lymphocytes, the NK cells lack the surface markers on their
cell membrane; they constitute about 10 – 15% of the circulating
lymphocytes. Once these cells activated, they can perform the functions of T
helper and cytotoxic T cells. They have cytoplasmic azurophilic granules
containing acid hydrolases.
Natural killer cells are able to attack and kill certain types of target cells, such
as: virus-infected cells, cancer cells, and transplanted cells without previous
stimulation.
N.B. Some researchers consider the natural killer cells as cytotoxic
lymphocytes, independent of thymus gland; they respond to foreign cells by
developing into lymphokine activated killer cells (LAK cells).

Major Histocompatibility Complex (MHC):

The major histocompatibility complex is a group of genes on the short arm of
chromosome 6 that code for several proteins located on cell surfaces to help

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the immune system to recognize the foreign substances. In humans, MHC is
also called human leukocyte antigen (HLA) system.
There are two general classes of the MHC, class I and class II. Both classes I
and II are integral membrane proteins present on the cell surface; they are
synthesized on polysomes and inserted on RER membranes. Proteins inside
the cell are broken into short fragments and displayed as peptide antigens by
MHC proteins on the surface. This helps the immune system to distinguish
between normal self and non-self foreign antigens.
Class I proteins are found in all nucleated cells and their function is to present
fragments of proteins that are synthesized inside the cell. Class I proteins join
the cytosolic proteins (virus infected cell) in RER to form class I MHC-
antigen complex. The complex is transferred through Golgi vesicles to the
surface of antigen presenting cells.
The peptide antigens presented in this manner are checked by killer T-cells,
which have receptors for the class I MHC proteins to identify the malignant
or infected cells to be attacked and destroyed.
Class II MHC proteins are found only in antigen presenting cells such as
macrophages that engulf foreign particles (bacteria). Class II MHC is
synthesized in RER, transferred to Glogi complex then to vesicles where they
fuse with endosomes containing engulfed antigens to produce class II MHC-
antigen complex to be exposed to the cell surface. These cells present peptide
antigens derived from such digested particles to helper T-cells, which have
receptors for class, II MHC proteins. The purpose of this process is to help
the immune system to stay in control and stop attacking the body’s own cells.
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Antigen-presenting cells (APCs):
These are a group of cells which are present in most tissues; they include
dendritic cells, macrophages, Langerhans cells, B-lymphocytes and
epithelial reticular cells of thymus. These cells have class II MHC
molecules on their surfaces; they are able to process antigen by digesting it
into small peptides or small molecular domains (epitopes) and present it on
the cell surface to be recognized by CD4 T-helper lymphocytes (CD8 T cells
interacts with class I MHC).
Dendritic cells arise from monocytes in the bone marrow; they are abundant
in T-lymphocyte zones of lymphoid tissue and in epidermis where they are
called Langerhans cells. In germinal centers of lymphoid follicles, there are
follicular dendritic cells with many processes that bind antigens on their
surfaces to be presented to B-lymphocytes.

Lymphoid Tissue
Lymphatic tissue occurs in many regions of the body as diffuse, dense or
nodular collections of lymphocytes (lymphatic nodules), or as lymphatic
organs covered by a definite capsule. Lymphatic tissue is involved with
lymphocyte production and immune responses.
The mucosa associated lymphoid tissue (MALT) is a non-encapsulated
aggregation of lymphoid tissue present in the mucosa of digestive (Peyer’s
Patches of the ileum and the appendix), respiratory and urinary systems.
Sometimes, the gut-associated lymphoid tissue is called GALT, and the
bronchiolar-associated lymphoid tissue is referred to as BALT.
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Newborn individuals, or animals reared in sterile conditions, possess very
few lymphatic nodules confirming that the lymphatic nodules develop in
response to foreign antigens.
Lymphatic organs are divided into:
 Primary or central: thymus and bone marrow
 Secondary or peripheral: lymph nodes, spleen, tonsils, solitary lymph
nodules, appendix and Peyer`s patches of ileum.
Lymphatic tissue consists of supportive reticular (collagen III) connective
tissue infiltrated with lymphocytes.
In lymphoid organs, except thymus, reticular fibrils are produced by
fibroblast-like cells with cytoplasmic processes called reticular cells. Small
and large lymphocytes are present in peripheral blood as well as in lymph.
The small lymphocytes re-circulate back and forth between blood and lymph,
bringing them into direct contact with foreign bodies to which they respond.
The immune response against lymph-borne antigen occurs in lymph nodes,
while the immune response to blood-borne antigen occurs in spleen.

Lymphatic Nodules:
Lymphatic nodules are found in all lymphatic organs (except thymus), and
in the loose connective tissue of many organs and in particular in the
lamina propria of the digestive tract, upper respiratory tract and urinary
passages.

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 Nodules are spherical structures (0.2-1mm diameter) without a
connective tissue capsule.
 Nodules are mainly composed of dense aggregates of small B-
lymphocytes.
 The cells at the nodule center are larger, with more cytoplasm, and
lightly-stained central area is known as the germinal or
germinative center.
 Nodules with a germinal center are sometimes described as
secondary nodules.
 The ring of small B-lymphocytes surrounding the germinal center is
referred to as the corona or mantle zone. If these small peripheral
lymphocytes are aggregated mainly at one edge, they are referred to
as a cap.
 The germinal center is the site of proliferation and differentiation
of B-lymphocytes; it includes activated B-lymphocytes (B-
immunoblasts), mitotic cells, plasma cells, dendritic reticular cells and
macrophages.

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 Thymus
Thymus gland is a lymphoid (lymphoepithelial) organ situated in the superior
mediastinum behind the upper part of the sternum. It is most active in young
people (at puberty) and involutes in old age but continues to produce
lymphocytes.
Unlike the rest of lymphoid organs which develop from mesoderm, thymus
gland has a dual (two) origin:
 Mesoderm (lymphocytes)
 Endoderm of third and fourth pharyngeal pouches (epithelial reticular
cells).
Thus thymus has unique features; it differs from the other lymphatic organs
in the following aspects:
1. Its stroma consists of a framework of connecting epithelial reticular cells
derived from endoderm, while typical reticular connective tissue present in
other lymphatic organs is mesodermal in origin.
2. Thymus gland has no lymphatic nodules; instead it is divided into
incomplete lobules whose lymphatic tissue is organized into cortex and
medulla.
3. Thymus gland does not have afferent lymphatic vessels; it contains efferent
lymphatic vessels in the walls of blood vessels and connective tissue septa.
4. There are no lymph sinuses in thymus.
5. Thymus does not filter the lymph; instead it is the site of T-lymphocytes
maturation.

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Structure of thymus gland:
Thymus gland is composed of stroma and parenchyma.
I. The stroma:
The gland is surrounded by a thin connective tissue capsule which sends
septa extending inwards dividing the gland into incomplete lobules and
forming short interlobular septa which end at the cortico-medullary junction.
These interlobular septa carry blood vessels which radiate into the substance
and branch at the cortico-medullary junction.
II. The parenchyma:
Thymus parenchyma is composed of lobules; each lobule consists of darkly-
stained cortex and lightly-stained medulla (it is not a germinal center).
The cortex:
The cortex is composed mostly of T-lymphocytes (called thymocytes), few
macrophages and epithelial reticular cells.
Just beneath the capsule, there is a continuous layer of epithelial reticular
cells. Deeper to this layer, a meshwork made by stellate reticular cells
which have branching stellate shape, acidophilic cytoplasm containing
bundles of tonofilaments, with large pale-staining oval nuclei ; their tapering
processes are joined to each other by desmosomes.
Some of the epithelial reticular cells contain large number (20-100) of
maturing T lymphocytes in their cytoplasm, and thus these epithelial reticular
cells are called thymic nurse cells (TNCs).
CD4ˉ and CD8ˉ cells, T cell precursors with no surface receptors, migrate
from bone marrow (liver in intrauterine life) and reside in the cortex where
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they are presented to self antigens bound to class I and class II MHC
molecules on APCs surface.
About 95% of the cortical T-lymphocytes that react to self antigens are
eliminated by the process of apoptosis or programmed cell death. If the self-
antigen reacting lymphocytes are not removed, an auto-immune disease will
develop.
The rest (small number) of T-lymphocytes matures, attains T cell receptors,
migrates to the medulla, enters the blood stream through the venules walls
and goes to nonthymic lymphoid organs where they reside in thymus-
dependant zones of the secondary lymphoid organs.

The medulla:
The medulla stains lighter because it contains more epithelial reticular cells
and few lymphocytes. It contains the characteristic acidophilic-stained
thymic or Hassall's corpuscles; they consist of concentrically arranged
epithelial reticular cells with keratin filaments in their cytoplasm. Epithelial
reticular cells degenerate and sometimes calcify; their function is unknown.
In young thymus, thymic corpuscles are small in size, whereas in old thymus,
the corpuscles increase in size and number.
Adrenocorticotropic hormones of anterior pituitary, adrenal cortex hormones,
male and female hormones, all accelerate thymus involution in experimental
animals.

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Blood supply of thymus:
1. Arteries enter the thymus through connective tissue capsule; arterial
branches follow the connective tissue septa deep into the gland.
2. Arterioles leave the septa to enter the parenchyma at the cortico-medullary
junction, break into capillaries which branch and anastomose forming
arcades which turn back and enter the medulla.
The capillaries, which supply the cortex (cortical) are impermeable to
proteins and prevent the circulating antigens to reach the site of T
lymphocytes maturation, forming blood-thymus barrier. This barrier is
composed of:
 The thymic cortical capillaries have nonfenestrated endothelial cells
with occluding junctions in between.
 Very thick basal lamina of thymic capillaries
This barrier receives an extra help from the continuous sheath of reticular
cells and the well-developed external lamina of reticular cells.
3. Post-capillary venules (high endothelial venules) in cortico-medullary
region with specialized cuboidal epithelium allowing lymphocytes in and out
of thymus.
4. The medulla is supplied by capillary branches of the arterioles at the
medulla-cortical border. The medullary (and septal) capillaries are
fenestrated and highly permeable.
5. Medulla is drained by venules which join forming veins.
6. Medullary veins pass through the septa and finally leave the gland through
the capsule.
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Functions of thymus gland:
1. Thymus is the site of production and maturation of T lymphocytes in an
antigen free environment.
2. The thymus reticular cells secrete thymolin which stimulates synthesis of
surface markers by immature lymphocytes.
3. Seceretion of thymic humoral factor, thymosin and thymopoietin which
promote differentiation of T-lymphocytes.
4. Overall control of immunity mechanisms.

Lymph Nodes
Lymph nodes are ovoid or bean-shaped organs, widely distributed in the
body. Important groups of lymph nodes occur in chains along the course of
lymph vessels in regions such as neck, axilla, groin, lung hila and para-aortic
areas. Lymph nodes are the only lymphatic organs to have an afferent
lymphatic supply. Lymph nodes are important for lymph filtration,
phagocytosis of particulate matters, production of lymphocytes, plasma cells
and antibodies which are added to the lymph leaving the nodes by the way of
efferent lymph vessels.

Structure of lymph node:

1. The hilum: Each lymph node has a hilum (hilus), which is an indentation
in the node through which arteries and nerves enter and veins and efferent
lymphatic vessels leave. The hilum may not be seen in every section.

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2. The capsule is made of dense connective tissue surrounding the lymph
node, and send trabeculae to the interior of lymph node.
3. The afferent lymph vessels:
 They penetrate the capsule at the convex side with an angle to enter
the subcapsular sinus.
 They are thin-walled lined with endothelium and contain valves to
prevent the backflow of lymph.

4. The cortex: is divided into outer and inner parts.

The outer part of the cortex lies immediately under the capsule; it contains
subcapsular sinus, diffuse lymphoid tissue, lymphatic nodules or follicles,
and trabecular sinuses.
The diffuse lymphoid tissue is mainly made up of T-lymphocytes, reticular
cells, macrophages and APCs.
The lymphatic nodules (primary nodules) are small round aggregations of
densely packed, small B-lymphocytes.
After exposure to antigen or infection, some nodules (secondary nodules)
develop light central areas called the germinal centers which show mitotic
figures. The germinal centers contain immature B-lymphocytes (B-
lymphoblast), dendritic follicular cells, plasma cells, and many macrophages.
Macrophages have acidophilic, vacuolated cytoplasm and an eccentrically
located nucleus. Sometimes, macrophages are filled with dark particulate
matters (dust or carbon).

239
The inner part of the cortex (deep cortex or paracortical area) is continuous
with its outer part. This inner part of the cortex is the thymus-dependant
zone and contains plenty of T-lymphocytes.

5. The medulla:
It is the inner zone of lymph node, radiating from the hilum. It contains the
medullary cords extending from the lymphatic nodules of the cortex and
composed of B lymphocytes, plasma cells and macrophages. The cords
branch and anastomose with each other; they are separated from each other
by dilated capillary-like structures called medullary lymphoid sinuses. These
sinuses are lined partially by reticular cells and macrophages. Reticular
fibers and cells cross the medullary sinuses giving the appearance of loose
networks.

Circulation of Lymph:
1. The lymph reaches the node through the afferent lymphatic vessels on the
convex side. Afferent vessels have valves which allow the passage of
lymph into one direction to the subcapsular sinuses.
2. From subcapsular sinuses, lymph passes to the intermediate sinuses
which run parallel to the trabeculae, into the interior of the node.
3. The lymph reaches the medullary sinuses where it slows down to allow
the uptake of foreign substances by macrophages and APCs.
4. The lymph is collected at the hilum by efferent lymphatic vessels which
have valves to prevent the backflow of lymph into the node.
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Blood supply of lymph node:
Lymph node is supplied by small arteries that enter the lymph node through
the hilum.
The arteries break into capillaries in the nodules where postcapillary venules
are formed and then small veins leave the node at the hilum. Postcapillary
venules or high endothelial venules (HEVs), in the paracortex, are special
types of vessels, lined with tall cuboidal cells rather than simple squamous
cells. Long-lived lymphocytes can recirculate many times by traveling
between the high cuboidal cells. The process by which lymphocytes return to
the lymph node through the wall of high endothelial venules is called
homing.

Functions of lymph node:

1. Clearing the lymph circulation from foreign particles because lymph must
cross at least one lymph node before entering the blood.
2. Filtering the lymph from particulate matters and antigens by macrophages
or APCs; all antigens are presented to T and B cells.
3. Formation of new lymphocytes (B cells and plasma cells in germinal
centers).
4. Production of immunoglobulins making the lymph leaving the lymph
nodes to the circulation is very rich with antibodies.

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 Spleen
The spleen is an organ which both forms and destroys blood cells; it is the
largest lymphoid organ.
General Structure:
The spleen is composed of stroma and parenchyma; its outer surface is
covered by mesothelium of visceral layer of peritoneum which is closely
attached to the capsule.
I. The stroma is composed of dense connective tissue capsule which sends
trabeculae into the parenchyma; it contains few smooth muscle fibers, elastic
fibers and a network of reticular tissue. At the hilum, the capsule sends
large connective tissue trabeculae carrying nerves, lymphatics and blood
vessels to and from the spleen.
II. The parenchyma (splenic pulp) consists of large number of lymphocytes,
reticular cells, blood cells, macrophages and APCs residing in the reticular
network; the cells are arranged as white and red splenic pulps.

The white pulp:

The white pulp consists of lymphatic sheaths around blood vessels and
lymphatic nodules. These nodules appear as white spots on the cut section of
a fresh spleen.
The lymphoid tissue surrounding the central arteries is called periarterial
lymphatic sheath (PALS); this is the thymus-dependant zone, and it is
composed mainly of T-lymphocytes.

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The lymphoid nodules and their germinal centers consist of B-lymphocytes
which are continuously exposed to blood-borne antigens.

The marginal zone is a transitional region surrounding the lymphoid nodule

and lies between the red and the white pulps; it includes splenic blood sinuses
with abundant blood antigens and loose lymphoid tissue containing B-
lymphocytes, plasma cells and many macrophages. In the marginal zone the
PALS becomes thinner and the central arteriole branches into straight
penicillar arterioles which have at their terminations thicker sheath of
reticular cells, lymphocytes and macrophages.

The red pulp:

The red pulp is composed of splenic blood sinusoids and splenic cords of
Billroth and loose network of stellate reticular cells and fibers. Its red color
is due to presence of large number of erythrocytes in blood sinusoids. The
splenic cords of Billroth occupy the spaces between the sinusoids and consist
of networks of reticular cells and fibers of collagen type III, T and B
lymphocytes, plasma cells, macrophages, platelets, erythrocytes and
granulocytes.

In the splenic sinusoids, the lining endothelial cells are unusually long
narrow and fusiform, with their long axes parallel to that of the sinusoids
which have discontinuous basal lamina. The endothelial cells are separated
by intercellular clefts and supported by transverse reticular fibers that encircle
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the blood sinusoids like hoops on a barrel without closing the endothelial
slits. This allows the blood cells to re-enter the circulation through these slits
and gives the spleen a unique filtering ability.
Functions of the red pulp:
 To produce immunoglobulins by plasma cells
 To filter and remove the worn-out cells and platelts by macrophages
 It acts as reservoir of erythrocytes and platelets

Blood circulation:
The spleen is supplied by the splenic artery which enters the spleen through
the hilum. Splenic artery branches into trabecular arteries which follow the
connective tissue trabeculae where they give smaller branches, and then leave
the trabeculae as central arteries to enter the parenchyma. These central
arteries are immediately ensheathed by T-lymphocytes, PALS.
Central arteries or the white pulp arteries are also referred as follicular
arteries; they have peripheral position in the follicle.
Sometimes the central arteries become arterioles, but still are called central
arteries; they give several branches to supply the surrounding lymphoid
tissue.
On leaving the white pulp, the central artery divides into numerous small
straight vessels called penicillar arterioles. These divide into smaller
terminal branches (capillaries) which are ensheathed by reticular cells,
lymphocytes and two or three layers of macrophages (ellipsoid or

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macrophage sheath). The mode of blood flow from the rterminal capillaries
into splenic sinuses is explained according two theories:
1. The closed circulation suggests that the blood remains always inside the
vessels and the terminal arterial capillaries open directly into the sinusoids
of the red pulp.
2. The open circulation suggests that the terminal arterial capillaries open at
their ends into the extracellular spaces of the red pulp allowing the blood
to circulate between the cells of the red pulp to reach the sinusoids. This
type of circulation is recommended by most researchers.
From sinuses, the blood drains into the pulp veins which enter the trabeculae
as trabecular veins. These trabecular veins drain into larger veins which, at
the hilum drain into the splenic vein.

Functions of the spleen:

 The spleen is highly active in production of antibodies
 It acts as filtering organ against microorganisms due to the reticular
structure of red pulp and the presence of macrophages
 It serves as temporary storage of platelets and erythrocytes. One third
of the body platelets are stored in the spleen and released as needed.
 The site of destruction of aged platelets and erythrocytes.
Macrophages are responsible for phagocytosis of worn-out
erythrocytes (may be called hemophages). Most of the iron

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 phagocytosed is restored to the circulation to be reused in production
of the new erythrocytes in the bone marrow.

** In the embryo, the spleen is a fully hemopoietic organ making all the
kinds of blood cells. In late fetal life, the spleen stops producing erythrocytes
and granulocytes, but continues to produce lymphocytes and monocytes.
Myeloid metaplasia or hemopoiesis in red pulp is a pathological condition
occuring in severe cases of anemia and leukemia.

Tonsils
The tonsils are aggregations of lymphatic nodules, incompletely encapsulated
and associated with epithelium.
The palatine, pharyngeal and lingual tonsils form a ring of lymphatic tissue
around the entrance to the throat. This protective tonsillar ring lies at the
oropharyngeal isthmus.

A. Palatine tonsils:
There are two tonsils; each of them is located at the lateral wall of
oropharynx. Palatine tonsils are covered on one side by stratified squamous
epithelium, nonkeratinized.
Ten to twenty tonsillar crypts extend from the surface of the tonsil into its
substance; their epithelial lining is continuous with the surface epithelium of
oropharynx. The crypts lumens contain desquamated cells, bacteria, live and

246
dead lymphocytes; the crypts appear as whitish purulent spots during
tonsillitis.
On the other side of the tonsil and opposite the surface epithelium, there is a
capsule of dense irregular connective tissue which serves to attach the tonsil
to the wall of oropharynx and prevents the spread of infection.
The capsule sends connective tissue septa into the tonsil separating the crypts
from one another and acts as barrier to prevent the spread of tonsillar
infections. Deep to the capsule, there are bundles of skeletal muscle fibers
and mucous glands.
The lymphatic nodules of the tonsil usually lie as a single layer just beneath
the epithelium surrounding each crypt.
Many of the nodules have germinal centers.
The afferent lymph vessels and lymph sinuses are absent and no cortex and
medulla pattern.

B. Pharyngeal tonsil:
Pharyngeal tonsil is a single one that lies in the superior posterior wall of the
nasopharynx. The covering epithelium is pseudostratified columnar ciliated
with goblet cells and some areas of stratified epithelium non-keratinized. In
the atrophic condition of the tonsil in adults, the ciliated epithelium is
replaced by stratified epithelium. It contains lymphoid nodules with germinal
centers, very thin capsule and has no crypts.
Enlargement or hypertrophy of pharyngeal tonsils results in obstruction of
nasal breathing and it is called adenoids.
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C. Lingual tonsils:
Lingual tonsils are located at the base of the tongue beneath the epithelium.
They are composed of small aggregations of lymphatic nodules covered by
stratified squamous epithelium some of them have germinal centers. Each
tonsil has a single crypt.
D. Aggregated nodules of Peyer`s patches:
The aggregated nodules of Peyer`s patches belong to a group of mucosa-
associated lymphoid tissue or Gut-Associated-Lymphoid Tissue (GALT).
They are located in the lamina propria and submucosa of small intestine
specially, opposite the attachment of the mesentery; the ileum has most of
the Peyer`s patches.
The lymphatic nodules appear by naked eye as dome-shaped area with no
villi.
 One Peyer`s patch may consist of 10-70 lymphatic nodules.
 The nodules originate in the lamina propria but may extend into the
submucosa disrupting the muscularis mucosa.
 When the nodules project into the luminal surface, the villi become
absent and the covering epithelium consists of M cells instead of
absorptive cells.
 The aggregated nodules are best developed in young children
occurring mainly in the lower part of the ileum.
 In adulthood and old age, the nodules undergo regression and
involution.

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 The Integumentary Syustem
The skin and its appendages that cover and protect the outer surface of the
body comprise the integumentary system.

The Skin
The skin is one of the largest organs in the body as it forms about 16% of the
body weight. It functions as a protective barrier against injury and
microorganism's invasion; it also protects against UV light by producing
melanin pigments. Skin prevents water loss; controls body temperature,
sensory perception through the receptors and formation of vitamin D3 by the
effect of the sun.
Skin is covered by keratinized stratified squamous epithelium, containing
keratinocytes (keratin-producing), melanocytes, Langerhans cells and
Merkel`s cells.
According to the thickness of the epidermis, skin might be divided into:
 Thick skin is found in soles and palms, where epidermis is 400
– 600 μm thick.
 Thin skin covers the body surface except soles and palms,
where epidermis is 75 – 150 μm thick.
In general, skin consists of two layers:
 Epidermis is an ectoderm-derived layer of stratified squamous
epithelium.

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  Dermis is the underlying thick layer of supporting connective tissue,
developed from mesoderm, highly vascular and contains sensory
receptors.
Subcutaneous tissue or hypodermis (beneath the dermis) does not make part
of the skin; it is made of loose connective and adipose tissue and called
superficial fascia. Hypodermis binds skin to the underlying tissue.

Thick Skin
Thick skin is present in the palms of hands and soles of feet. It is
characterized by having a thick layer of stratum corneum. The skin of the
finger tips shows distinctive pattern of arches and loops of ridges and
grooves, called dermatoglyphics. These patterns make the bases of specific
individuality of fingerprints.
Epidermis
Epidermis of thick skin shows downward folds called epidermal ridges or
rete ridges lying between the dermal pegs or papillae of the dermis. It
consists of 5 layers of cells called keratinocytes which are responsible for
keratin formation and 3 less abundant cell types (melanocytes, Langerhans
cells and Merkel`s cells).
I. Keratinocytes form the keratinized stratified squamous epithelium and are
arranged in 5 layers:
1. The stratum basale is a single layer of simple columnar or cuboidal cells
resting on the basement membrane at the junction of dermis with epidermis.
The lateral and apical sides of the cells are joined by desmosomes.
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Hemidesmosomes are present between the cell bases and the basal lamina.
The cells are basophilic, show mitotic figures and contain intermediate
keratin filaments which bincrease in number as the cells move upwards.
Together with the stratum spinosum, cells of stratum basale (stem cells) are
responsible for epidermal rgeneration or renewal (every 15-30 days).
Melanocytes are located in this layer; they have pale or clear cytoplasm and
round dark nuclei.
2. The stratum spinosum or prickle cell layer consists of several layers of
cuboidal to polyhedral cells lying immediately above the stratum basale.
The cells show basophilic cytoplasm, central round nuclei with prominent
nucleoli and fine processes filled with keratin filaments. These processes
are firmly bound together by desmosomes which appear on the cell surface
as spines or prickles. The keratin filaments appear under the light
microscope as tonofilaments which maintain the cell cohesion and resist the
mechanical forces.
Malpighian layer consists of stratum basale and stratum spinosum, it is
responsible for skin renewal by epidermal stem cells.
3. The stratum granulosum consists of 3 - 5 layers of flattened polygonal
cells. Their cytoplasm exhibits coarse basophilic keratohyaline granules
made of proteins rich in phosphorylated histidine and are not limited by
membranes.
By EM, the granulosa cells appear to have lamellar granules which consist of
alternating electron-dense and electron lucent lamellae of lipid bilayers. The

251
granulosum cells release their contents by exocytosis into the intercellular
spaces, to form a sealing barrier against microorganisms' penetration.
4. The stratum lucidum is a thin translucent layer, made of flattened, very
thin, pale staining eosinophilic cells. This layer is present only in thick skin;
it lies between the stratum granulosum and stratum corneum. Nuclei and all
cell organelles are not seen; the cytoplasm is filled with densely-packed
keratin filaments.
5. The stratum corneum consists of 15 - 20 layers of flattened non-nucleated
keratinized cells whose cytoplasm is filled with keratin filaments. After
kiratinization, the cells consist of amorphous substance and thick plasma
membrane and are called horny cells. The dead cells are continuously shed
off and replaced from below.
The cornified layer of epidermis is composed of soft keratin, in contrast to
hard keratin found in nails and hair.
In psoriasis, there is high regeneration rate and increased number of the cells
of stratum basale and stratum spinosum resulting in great thickness of
epidermis.

II. Melanocytes:
Melanocytes are melanin producing cells. Melanin is one of the major
pigments, beside carotene and hemoglobin affecting skin color.
Melanocytes develop from neural crest (ectoderm) of the embryo; they are
located between the cells of stratum basale and basement membrane and in
the hair follicles.
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The cell bodies of melanocytes are rounded with radiating branching
processes called dendrites (extentions), running between keratinocytes of
stratum basale and spinosum. The processes end by invaginations between
the cells. By EM, the cells appear pale with numerous mitochondria, few
RER, well-developed Golgi complex and hemidesmosomes between the cells
and basal lamina.
Function of melanocytes: Melanin synthesis.

Melanin synthesis:
Tyrosinase Tyrosinase Tyrosinase
Tyrosine →→→ DOPA →→→ DOPA quinine →→→ Melanin
1. Melanin synthesis starts by making electron-dense fine protein granules at
the periphery of a small vesicle, by the activity of tyrosinase enzyme on the
amino acid tyrosine resulting in formation of dihydroxyphenylalanine
(DOPA). Dopaquinone is formed by the action of tyrosinase enzyme on
DOPA.
2. Melanin is deposited in the vesicles which are called melanosomes.
3. Melanosomes show periodicity which disappears by increased formation of
melanin.
4. The mature melanin granule is ellipsoid in shape, 1x0.4 μm, fills the
vesicle completely.
Melanin is synthesized in melanocytes, migrates to its processes and then
transferred to keratinocytes cytoplasm of stratum basale and spinosum which
contain more melanin pigments than melanocytes.

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Melanin synthesis is promoted by melanocyte stimulating hormone (MSH) of
pituitary gland.
Exposure to sun helps to increase the already present pigment and sitimulate
melanocytes to synthesize extra melanin.
Pheomelanin is the pigment present in red hair, while eumelanin is the dark
brown pigment of the dark hair.
The pattern of distribution of melanocytes among keratinocytes is called
epidermal-melanin unit i, e the number of melanocytes per unit area.
Malignant melanoma is highly invasive malignant tumor of melanocytes

III. Langerhans cells:

Langerhans cells are stellate cells with dark staining nuclei, present mainly in
the stratum spinosum. They lack the bundles of keratin filaments and have
no desmosomes on their surface. They originate in bone marrow and carried
to the skin by blood. They are APCs as they take antigen and present it to T-
lymphocytes; hence they play an important role in immune responses.

IV. Merkel`s cells:

Merkel cells are present in epidermis of thick skin. Their cytoplasm contains
small dense granules.
The free nerve endings terminate at the bases of these cells by making disc-
shape expansion. They function as mechanoreceptors for touch sensation.

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 Dermis
The dermis is made of connective tissue that supports the epidermis and binds
it to hypodermis (subcutaneous tissue). It is rich in elstic fibers, blood and
lymph supply. In some areas, blood can pass directly through arteriovenous
shunts (anastomoses).
The dermis contains the epidermal deravatives such as hair follicles, sweat
and sebaceous glands.
The dermis is composed of papillary and reticular layers.
1. The papillary layer has many projection (dermal papillae) which
interdigitate with the epidermal pegs or ridges to strengthen dermal-
epidermal junction.
This layer is composed of loose connective tissue; it contains connective
tissue cells, collagen and elastic fibers, blood and lymphatic vessels, and
tactile corpuscles of Meissner. Fine collagen fibrils, anchoring fibrils,
binding the dermis to basal lamina of stratum germinativum of epidermis
2. The reticular layer contains dense irregular connective tissue (collagen
type I), blood vessels, nerves and sensory receptors (Krause end bulb, Ruffini
and Pacinian corpuscles), and ducts of merocrine (eccrine) sweat glands.

Thin Skin
Thin skin covers the whole body except palms and soles.
In thin skin, epidermis in general, the stratum corneum, granulosum and
spinosum are thinner than those of thick skin. No stratum lucidum in thin
skin.
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 Skin appendages
All skin appendages are derived from epidermis; they are:
 Hairs
 Nails
 Eccrine (merocrine) and apocrine sweat glands
 Sebaceous glands which open into hair follicles
Hairs:
The hairs are keratinized structures developing from hair follicles which are
epidermal derivatives; they develop by downgrowth of epidermis into the
dermis. Hair might be terminal (scalp and male beard hairs) or vellus, short
fine hairs covering the whole body surface. Hairs present all over the body
skin except:
 Palms  Glans penis
 Soles  Clitoris
 Lips  Labia minora
Hair color and texture differ according to melanin content, the age, sex, race,
and the area of the body.
The hair is composed of the bulb, the root (the part of the embedded in
skin) and the hair shaft (the part protruding above the skin surface).
The hair follicle has a bulb-like lower expansion, with a dermal papilla at
its base. The papilla contains networks of capillaries which nourish the hair
follicle, and it is covered by cells which form the hair root which develops
into the hair shaft.

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The epithelial cells of the bulb act as the stratum basale (germinativum) of
the skin; its central cells, differentiate into large vaculated moderately
keratinized cells forming the medulla of the hair.
The cells at the lateral side of the papilla differentiate into heavily keratinized
hair cortex, and the next layer of cells differentiates into the cuticle of the hair
which is made of cuboidal cells that change into columnar cells. High up in
the hair, the cuticle cells become flattened and heavily keratinized.
The outermost layer of cells, the peripheral epithelial layer, differentiates into
internal and external root sheath.
The internal root sheath covers the beginning of the hair shaft then
degenerates at the level of opening of the sebaceous glands.
The external root sheath is thin at the level of dermal papilla; it is composed
of one layer of cells which corresponds to stratum basale cells. Near the
surface of the skin, the external root sheath is contineous with the layers of
epidermis.
Melanocytes are located between the papilla and the epithelial cells of the
hair root; they are responsible for the hair color.
The hair follicles are separated from the dermis by a noncellular hyaline layer
called glassy membrane, formed by thickening of the basal lamina. The
dermis around the hair follicle forms a dense connective tissue sheath which
is connected to the papillary layer of the dermis by smooth muscle bundles
called arrector pili muscle.
Contraction of arrector pili muscle causes erection of the hair shaft and
depression on the skin surface (goose flesh).
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Keratinization of the hair follicle is different than that of epidermis in the
following:
1. Keratin of epidermis is soft and loosely attached and desquamates
continuously, while the hairs keratin is hard and compact.
2. Keratinization of epidermis is continuously over the entire surface, while
in the hair keratinization is intermittent and involves the hair root only.
3. Cell differentiation in epidermis results in one layer allover the skin
surface, while in the hair follicle, differentiation results in different cell types.
Cell division and cell differentiation in the hair follicles are under the effect
of undrogens.

Hair growth and replacement:

Hair growth is not a continuous process; it passes through three stages.
Anagen is the stage of growth, catagen is the stage of cessation of growth and
telogen when the hair follicle atrophies and finally is lost. There is a constant
gradual hair loss and replacement.
Scalp hair has the longest duration, two to five years, while eyebrows last
three to five months. When the hair is about to shed, proliferation of the
cells above the papilla slows down and then stops.
The bulb develops into solid club-shaped mass which fuses with the cells of
the inner root sheath. Finally, the club hair becomes detached and slowly
shifted towards the surface. Formation of new hair starts with proliferation of
the cells of the external root sheath.

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Nails:
The nail is resting on the dorsal surface of the fingertip. It is composed of a
root (containing the nail matrix) and a plate of keratinized epithelial cells
resting on the nail bed.
The proximal part of the nail is called the root or matrix and is covered with
skin, called nail fold (eponychium is formed of stratum corneum of the free
edge). Nail matrix is responsible for the growth of the nail plate.
The skin lying under the nail plate is called the nail bed and composed of
stratum basale and stratum spinosum only.
The nail plate is composed by the division of cells of the matrix; it is made
of hard keratin and its free end is worn out.
Lunula is white opaque crescent present at the nail proximal end.
Sebacious Glands:
Sebacious glands are simple alveolar, holocrine exceretory glands. They are
associated with hair follicles; their ducts open on the upper part of the hair
except in certain areas like glans penis, clitoris and lips where they open
directly on the skin surface. Their secretion is called sebum, an oily
substance that keeps the hair shiny, and might have an antifungal and
antibacterial effect but does not prevent water loss.
Sebacious glands begin functioning at puberty, controlled by testosterone in
man and by ovarian and adrenal androgens in women.
Structure:
The secretory part of a sebacious gland consists of several acini which open
into a short duct. The acini consist of a basal layer of undifferentiated
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flattened epithelial cells which rest on the basal lamina. The acinar cells
contain fat droplets in their cytoplasm with small gradually shrinking nuclei
as the fat droplets fill the cytoplasm. When the cells burst, sebum is
produced on the skin surface and along the hair follicles. Sebum is a
complex mixture of lipids including triglycerides, waxes, squalene,
cholesterol and remnants of dead cells (holocrine).
Sebacious glands start to function at puberty under the effect of androgens.
Acne occurs at puberty due to chronic inflammation of obstructed sebacious
glands.

Sweat Glands:
They are widely spread in both thick and thin skin except in some areas such
as nail bed, ear drum and glans penis.
They are simple coiled tubular glands, located deeply in the dermis.
Sweat glands are of two types:
1. Eccrine (merocrine) sweat glands have cholinergic innervation
2. Apocrine sweat glands have adrenergic innervation

The merocrine (eccreine) sweat glands:

They are present allover body except ear drum, glans penis and nail bed.
They are simple coiled tubular glands whose ducts open at the skin surface
and are smaller in diameter than their secretory parts.
The secretory portion has large diameter (0.4 mm) and is made of acini
which are surrounded by myoepithelial cells which help to discharge
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secretion when they contract. The cells lining the secretory portion (acinus)
are of two types:
1. Dark cells are pyramidal in shape lining most of the lumen with
glycoprotein secretory granules at their apices. The basal surface of these
cells does not reach the basal lamina.
2. Clear cells have no secretory granules; their cell membrane has numerous
invaginations characteristics to ion-transport cells.
The excretory ducts are coiled and lined by two rows of darkly stained
stratified cuboidal epithelium. They open on the skin surface.
Function: Secretion of sweat and elimination of metabolism byproducts.
The secreted sweat is not a viscous solution and contains very little amount of
prteins. Sweat is an ultrfiltrate of bood plasma, made maily of water, sodium
chloride, urea, ammonia and uric acid. The duct cells are responsible for
sodium reabsorption to prevet its excessive loss.

The apocrine sweat glands:

These glands are present in certain areas such as: axilla, areola of breast,
pubic and anal regions; they start functioning at puberty due to sex hormone
influence. They are wider than the eccrine sweat glands (3 -5 mm in
diameter). They are located deep in the dremis and hypodermis and their
ducts open into the hair follicle.
Apocrine sweat glands secrete viscous, odorless secretions which later
acquire a bad smell due to bacterial decomposition.

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** The glands of Moll of the eyelids and ceruminous glands of the ear are
modified sweat glands.

The blood and lymphatic supply of the skin:

Skin is a highly vascular organ; it receives its arterial supply through two
plexuses:
 Subpapillary plexus is located between the papillary and
reticular layer of dermis
 Cutaneous plexus lies between the dermis and hypodermis.

The skin has three venous plexuses, the subpapillary, and the cutaneous and
a third venous plexus lying in the middle of the dermis.
Beside arterial and venous plexuses, skin is rich in arteriovenous anastomses
and glomera (plural of glomus, a special type of A-V anastomosis found in
finger nail beds and ears).
Lymphatic vessels start as blind sacs in the dermal papillae and converge
into two plexuses which accompany the arterial cutaneous and subpapillary
plexuses.

Nerve supply of the skin:

Skin is very rich in sensory innervation to receive sensory stimuli from the
environment.
A. Non-encapsulated nerve emdings:

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  Free nerve endings for pain, temperature and touch, these are
found in the epithelium.
 Peritrichial nerve plexus around the hair follicles
 Merkel's disk: for touch, present in the epithelium.
B. Encapsulatednerve endings:
 Tactile corpuscles of Meissner present in dermal papillae
 Krause end bulb present in dermis
 Ruffini corpuscles present in hypodermis.
 Pacinian corpuscles present in hypodermis, most numerous in
palms and soles. They are deep pressure receptors.

Functions of the skin:

1. Protection against trauma, microorganisms, water loss and ultraviolet rays
2. Regulation of body temperature and blood pressure
3. Sensory perception of tactile, thermal and painful stimuli
4. Excretion of water, fat and metabolic byproducts
5. Formation of Vitamin D3 by the effect of sunshine or ultraviolet light on
the provitamin 7-dehydrocholesterol present in the skin.

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