LAB

Exercise 1

Dissolution

Results
Which substance(s) dissolved?
Which substance(s) did not?
How can you tell?

Filtration

Results
Which substances passed through the selectively permeable disk?
Which substances did not?
Why do some substances pass through the filter paper while other substances do not?
Diffusion
Methodology
Prepared 5% agar and poured onto 6 petri dishes in equivalent amounds
After agar was set, a well was created in the center of each petri dish
Prepared dyes
methylene blue
100%
50%
Congo red
100%
50%
Filled wells with dye
A
Petri dish 1: 100% methylene blue
Petri dish 2: 100% Congo red
B
Petri dish 3: 100% methylene blue
Petri dish 4: 100% Congo red
C
Petri dish 5: 50% methylene blue
Petri dish 6: 50% Congo red
Setup
A
Petri dish 1: Room temperature
Petri dish 2: Room temperature
B
Petri dish 3: Refrigerated
Petri dish 4: Refrigerated
C
Petri dish 5: Room temperature
Petri dish 6: Room temperature
After finishing set-up, time was recorded
Petri dishes were checked 10 times within 3 days to observe for diffusion and the following were
done in a table:
time and date were recorded
measured distance from the edge of well to the distance of the edge of diffusion (in mm)
Calculations and Analysis
Rate of linear diffusion was computed in mm per hour
Created line graph comparing the three setups
X-axis: time in hours
Y-axi: rate of linear diffusion
Results
Factors affecting diffusion
Temperature
increasing temperature increases movement, allowing diffusion to take place
all forms of motion is inuenced by heat energy
heat has the ability to cause random motion in microscopic particles such as
atoms and molecules
increase in temperature, increases rate of diffusion
Molecular Weight/Particle Size
Congo red (696.68) diffused faster than methylene blue (319.86)
methylene blue should have diffused faster because of its smaller molecular
weight and particle size
results reflect that factors other than molecular weight had more bearing on
diffusion (see membrane permeability)
increase in molecular weight, decreases rate of diffusion
Membrane Permeability
agar = 5% agar + 95% water
Congo red is water soluble
methylene blue is slightly soluble in water
Since agar is mostly water, and methylene is more hydrophobic, it would diffuse
slower despite having a smaller molecular weight
increase in membrane permeability, increases rate of diffusion
in this case, increase in water solubility, increases rate of diffusion
Concentration
100% concentrated dyes diffuse faster than less concentrated ones
100% concentrations of methylene blue and congo red have 0% water
5% agar has 95% water
dyes are more concentrated than agar (agar has more water)
well containing dye is concentrated so dye moved to regions with less
concentration and more water (moved away from the well and toward
the agar)
50% concentrated dyes contain 50% water
diffused slower into the 5% agar containing 95% water
increasing concentration, increases rate of diffusion

Osmosis
Concept of osmosis
diffusion of water through a semi-permeable membrane
water moves from lower solute concentration to higher solute concentration
water moves from higher solvent concentration to solvent water concentration
movement occurs until equilibrium is reached
solutions
hypotonic
solute concentration is higher outside
results in cell bursting
hypertonic
solute concentration is higher inside
results in cell shrinking
isotonic
solute concentration is equal
equilibrium
Methodology
cellophane represented the semipermeable membrane
Results
increase in solute concentratation, increases fluid displacement
increase in solute concentration is equivalent to the increase in concentration gradient
higher concentration gradient, slows down equilibrium

Hemolysis
Concepts
hypotonicity
higher concentration of solute outside cell, a lot of water inside cell
cell tends to burst
principle behind hemolysis
hemolysis
clear solution = complete hemolysis
isotonic coefficient
molarity at which non-electrolyte (glucose) completely hemolyzed rbc molarity at which
electrolyte (sodium chloride) completely hemolyzed rbc
amount of salts to be added to distilled water to make the solution isotonic for rbc
degree of dissociation
the fraction of the original solute that has dissociated
Methodology
Results
hemolysis was observed in setups with:
calcium chloride (CaCl2)
potassium chloride (KCl)
hemolysis decreases as dilution decreases
increase in dilution causes increase in hypotonicity, increasing hemolysis
2 ions completely dissociated when the isotonic coefficient is 3

Fermentation
Concept: The formation of gas by fermentation produces pressure in the tubes causing the liquid to be pushed.

Methods:
Fermentation tubes were given mixtures:
o A - 15 ml of 10% glucose solution
o B - 7.5 ml of 10% glucose solution and 7.5 ml yeast suspension
o C - 15 ml sucrose solution
o D - 7.5 ml of 10% sucrose solution and 7.5 ml yeast suspension
o E - 15 ml yeast suspension

The rate of fermentation was measured as the amount of gas displaced in the tube (measured by cm using
ruler) and were measured at different time intervals of 10 mins, 15 mins, 30 mins, 1 hr, 2 hrs, and 24 hrs.
o A and C did not ferment as there was no yeast
o Change in the pressure of the tubes results when yeast ferments sugars. Production of carbon dioxide pushes the
solution. Foaming was also produced due to carbon dioxide.
o E should not have fermented since supposedly no sugars should be found.
*Error: Residual sugars were found. Tubes were not cleaned properly
o B (containing glucose) fermented faster than D (containing sucrose) because yeast fermentation converts simple
sugars (glucose) to ethanol and carbon dioxide. A disaccharide, has to be first degraded into its monosaccharide
units before it proceeds to fermentation.
o The fast rate of fermentation of E must have occurred since E contained the most amount of yeast.

Additional test for acidity. 5 drops of phenol red was added then gently shook. The color change of the solution
was:
o A - Transparent yellow
o B - More opaque yellow separated from a darker brown liquid
o C - Transparent yellow
o D Most opaque yellow
o E - More opaque yellow separated from a darker brown liquid
Phenol red is a pH indicator that turns red at or above pH 7 (alkaline) and turns yellow at a pH lower than 7
(acidic). All setups produced a yellow color indicating their acidic nature. Carbon dioxide from fermentation reacted to
components of the solution which yield to acids such as carbonic acid.




Exercise 2

Brainstem Reflexes
Methodology
Pupillary Reflex
shine light on one eye and observe pupil of that eye
Consensual Pupillary Reflex
shine light on one eye and observe pupil of that other eye
Ciliospinal Reflex
pinch neck and observe reaction of eyes
Corneal Reflex
suddenly touch cornea of eye and see what happens
Orbicularis Oculi Reflex
surprise person by flashing light on their eyes and observe reaction
Auditocephalogyric Reflex
surprise person with loud sound and observe reaction
Gag Reflex
prod pharyngeal region of neck and see what happens
Results
Pupillary Reflex
result: pupil constriction
pupils role is to allow light to enter retina
constriction happens when there is too much light to protect photoreceptors
anatomic components
CN II
Optic nerve
sensory nerve for eye region: responsible for detecting stimuli
CN III
Oculomotor nerve
motor nerve: responsible for effector
Consensual Pupillary Reflex
result: pupil constriction
though unexposed to light, also constricts so response is consensual
optic tract conducts visual information from both eyes and the pretectal area
projects bilaterally to both Edinger-Westphal nuclei
pretectal area
narrow, transversely oriented rostral zone of the
mesencephalic tectum
contains several nuclei that receive fibers from the optic tract
has bilateral efferent connections with the Edinger-Westphal
nucleus of the oculomotor nuclear complex by way of which it
mediates the pupillary light reflex
Edinger-Westphal nuclei
contains preganglionic parasympathetic (visceromotor)
neurons whose axons end in the ciliary ganglion
Input to the Edinger-Westphal nucleus arises from a cell group
called the pretectum, a cell complex that receives retinal input
and is part of the pathway involved in reducing the size of the
pupil upon light stimulation of the retina
anatomic components
CN II
Optic nerve
sensory nerve for eye region: responsible for detecting stimuli
CN III
Oculomotor nerve
motor nerve: responsible for effector
Ciliospinal Reflex
result: pupil dilation
eyes dilate in response to pain: defense against noxious stimuli
person becomes alert to sensations of pain in neck
anatomic components
CN II
Optic nerve
sensory nerve for eye region: responsible for detecting stimuli
CN III
Oculomotor nerve
motor nerve: responsible for effector
Corneal Reflex
result: blinked
blinked in response to tactile stimulation as defense mechanism
anatomic components
CN II
Optic nerve
sensory nerve for eye region: responsible for detecting stimuli
CN III
Oculomotor nerve
motor nerve: responsible for effector
Orbicularis Oculi Reflex
result: blinked
defense to noxious stimuli
anatomic components
CN II
Optic nerve
sensory nerve for eye region: responsible for detecting stimuli
CN III
Oculomotor nerve
motor nerve: responsible for effector
Auditocephalogyric Reflex
result: head and eyes turned toward direction of stimulus
defense mechanism: concept of arousal from psychology
arousal: type of bodily energy which primes or prepares us for emergency action
anatomic components
CN IV
Trochlear nerve
motor nerve: responsible for eye movement
directed eye to seek stimulus (loud sound)
CN VIII
Vestibulocochlear nerve
sensory nerve: for hearing and balance
CN XI
Spinal accessory nerve
motor nerve: responsible for head movement
Gag Reflex
result: gagged
defense mechanism: discourage organism from eating incompatible substances
(allergenic, toxigenic)
anatomic components
CN IX
Glossopharyngeal nerve
sensory nerve: responsible for detecting stimuli in pharyngeal area
CN X
Vagus nerve
sensorimotor nerve: responsible for sensory and motor functions in
viscera
triggers regurgitation

Stretch (Deep Tendon) Reflexes

Spinal Cord Reflexes

Reflex
basic unit for behavior is the reflex
generally occur quickly and are the result of excitation of
sensory receptors
afferent nerve fibers
integration by the central nervous system
transmission of motor impulses over efferent (motor) nerve fibers
excitation of an effector organ /tissue.
spinal cord may act as the primary integrator and source of a motor nerve response (spinal reflex) or as a
modulator of motor responses when the brain or higher regions of the spinal cord are involved
integrating and modulating activity on reflex activity is observed best when the controlling influences of the
brain are removed (why pithing is needed)

Single pithing
ablation of brain
insert the scissors so they are back against the joint between the upper and lower jaws before cutting
effectively breaks all connections between the brain and the spinal cord, it therefore eliminates any influence
the brain could have on reflex activity

Double pithing
ablation of spinal cord and brain
destroy the spinal cord by insertion of a mental dissection probe/needle into the open vertebral canal by the
single pith


Muscle Tonus

Electroencephalography (EEG)

Reflex and Reaction Time

Compound Action Potentials in Frog Sciatic Nerve
large peripheral nerves
bundles of thousands of individual axons encolosed in epinerium
ex. vagus, sciatic, ulnar nerves

epinerium
loose connective tissue sheath

fascicles
smaller bundles of axons within epinerium
each of which is also encased in perineurium

perineurium
more structured epithelial sheath

endoneurium
each individual axon is surrounded by a very thin individual connective tissue sheath

toads sciatic nerve
consists of only a single bundle of fibers, surrounded by the perineurium and loose epineurium

Compound nerve
ex. sciatic nerve
typically contains efferent ( and motor axons, and post-ganglionic autonomic axons) and afferent
(sensory) axons
individual axons vary in diameter, myelination, excitability, threshold and conduction speed.

Compound Action Potential
algebraic sum of many individual all-or-none action potentials arising more or less simultaneously in a
large number of individual axons in a large compound nerve does not occur naturally
elicited experimentally or clinically by stimulating the whole nerve with extracellular stimulating electrodes
and is recorded by means of extracellular recording electrodes, which measure the summed electrical
response of all the excited axons in the nerve
properties of the CAP: threshold, amplitude, duration, conduction velocity are determined by the type and
number of individual axons which are recruited (excited) by the stimulus
number and type of axons excited depend on the intensity of the stimulus
demonstrated by extracellular recording from many axons, is a graded response whose magnitude
increases with the intensity of stimulation because different axons have different thresholds of excitation
largest axons have the lowest threshold of excitation i.e., they are the most excitable. (Thus, in terms of
excitability, A axons>A>A>A>B>C)
at low stimulus intensities, only the largest axons are activated, but as the stimulus intensity is raised in
steps, more and more smaller axons are progressively recruited
shape has the biphasic and unsymmetrical characteristics
biphasic: each has a positive and negative component
biphasic nature of the CAP is due to the fact that the CAP is recorded with a pair of extracellular
electrodes (bipolar recording)
when the nerve is inactive, there is no potential difference between the electrodes, and the trace is
at baseline
When the CAP arrives at the indifferent electrode (1), the electrode becomes transiently negative to
the recording electrode (2), the potential difference between the two is detected and amplified by
the differential amplifier, and the polarity is stated in terms of the sign of charges in the recording
electrode (3), and the trace is displayed as an upward deflection on the screen
When the CAP lies between the indifferent and recording electrodes there is no potential difference
between the two electrodes and the trace returns to baseline
As the CAP moves down the nerve, the recording electrode becomes relatively negative and the
potential difference between the electrodes is now seen as a downward deflection on the screen.

Single Action Potential
demonstrated by intracellular recording from a single axon
all-or-none response: that is, under normal circumstances, the amplitude of the response does not change
with stimulus intensity (above threshold)
biphasic: each has a positive and negative component
The classic biphasic action potential recorded inside a single axon with a single intracellular
electrode consists of an initial (positive) depolarization due to a transient increase in Na+
permeability, followed by a negative phase of hyperpolarization
hyperpolarization is due to a prolonged increase in K+ permeability
positive and negative phases of the intracellular action potential are thus generated by a sequence
of selective membrane permeability changes, which result in depolarization and hyperpolarization
of the cell membrane.
conduction velocity of individual action potentials increases with the axon diameter
action potentials of the largest axons will reach the recording electrodes first

Stimulus artifact
display initially will show only a brief biphasic deflection.
results from the instantaneous spread of passive electrical current from stimulating to recording electrodes
by the electrolyte on the surface of the nerve

Initially, at low stimulating voltages, there will be no CAP.
CAP: as stimulus strength (voltage) is gradually increased a small deflection appears to the right of the SA

Threshold Stimulus Voltage
by carefully raising and lowering the stimulating voltage, find the smallest discernable deflection

Maximal Stimulus Voltage
Continue to increase the stimulating voltage, observing the changes in shape and magnitude of the CAP
until further voltage increase no longer increases the amplitude of the CAP

Refractory Period
during the action potential, a second stimulus will not produce a second action potential.
we can give the sciatic nerve two stimuli successively, and the intensity of the two stimuli is identical
If the first stimulus can produce an action potential, then the second stimulus can also produce action
potential, if the interval of the two stimuli exceeds the refractory period of the action potential arising from the
first stimulus
At the beginning, the interval of the tow stimuli is very long and exceeds the refractory period of the
first action potential arising from the first stimulus considerably. Under this circumstance, both the
two stimuli can produce action potential
Then, we minish the interval gradually, at first, we can record two action potential
When the internal is short enough that it equals the refractory period of the first action potential, the
second stimulus cant produce action potential, and the interval of this two stimuli is the refractory
period of the action potential.

Conduction Velocity
d/t --where d = the conduction distance; --and t = the conduction time.


Study Questions
1. How does a CAP differ from a single action potential?
A compound action potential is the sum of multiple axons in a nerve firing, while a single action potential is
generated by just one axon.
single action potential (recording from a single axon) is an all or none response
under normal circumstances, the amplitude of the response does not change with stimulus intensity
conduction velocity of individual action potentials increases with axon diameter
action potentials of the largest axons will reach the recording electrodes first

compound action potential (recording from many axons) is a graded response
magnitude increases with intensity of stimulation
this is because different axons have different thresholds of excitation
largest axons have the lowest thresholds of excitation and are the most excitable
at low stimulus intensities, only the largest axons are activated
as stimulus intensity is raised in steps, more and more smaller axons are progressively recruited


2. Action potentials are said to be all or none responses. Why does the frog sciatic nerve give a graded
response?
The frog sciatic nerve is made up of different axons, each with their own threshold. Some are higher than
others while some are lower. Therefore, as you increase the voltage, more and more of them fire, giving a
seemingly graded response
This graded response phenomenon illustrates the differences in threshold that exists among the different
sizes of fibers that make up the nerve. Remember, you are recording from a nerve, a large bundle of
neurons, each with a different threshold. If the stimulus voltage is increased slowly and smoothly, you may
observe discrete jumps in the amplitude of the compound action potential as different threshold classes of
nerve fibers are recruited. As you increase the amplitude more neurons reach their threshold and
contribute to the increase in size of the compound action potential. Eventually, as the stimulus voltage is
increased, a point will be reached when the wave form of the action potential stops changing. At this point all
the fibers in the nerve able to respond to the stimulus are being stimulated. This is a maximal response.

3. What was the smallest voltage required to produce the maximum (largest) CAP? What proportion of the
nerve fibers were excited to produce this maximal response?
To produce a maximum response, the whole nerve must be stimulated.
If the stimulus voltage is increased slowly and smoothly, you may observe discrete jumps in the amplitude of
the compound action potential as different threshold classes of nerve fibers are recruited. As you increase
the amplitude more neurons reach their threshold and contribute to the increase in size of the compound
action potential. Eventually, as the stimulus voltage is increased, a point will be reached when the wave form
of the action potential stops changing. At this point all the fibers in the nerve able to respond to the stimulus
are being stimulated. This is a maximal response.

4. In this exercise, you examined the effect of increasing stimulus intensity on the nerve. What other stimulus
parameter might also affect the nerves tendency to generate a CAP?
Frequency and duration can affect the nerves tendency to generate a CAP; frequency in that if theres the
right amount of time between them, their effects can be additive and trigger a CAP; duration in that even a
weak stimulus can potentially trigger a CAP if kept up long enough

5. Explain the difference between the relative and absolute refractory periods.
No action potentials can be fired during the absolute refractory period since too many Na+ channels are
voltage inactivated from the last action potential being fired. However, during the relative refractory period,
enough Na+ channels have opened back up to allow another action potential to be fired, though a stronger
stimulus than normal will be required.

6. Briefly describe the cellular events responsible for the refractory period. (Hint: Discuss the mechanism of
repolarization.)
After an action potential is fired, a large fraction of the cells Na+ channels are voltage inactivated,
preventing a second action potential from being fired. This is the absolute refractory period. The cell then
releases K+, repolarizing the membrane and causing the Na+ channels to open back up. Between the
absolute refractory period and reaching the normal resting membrane potential comes the relative refractory
period, which is explained above.

7. Based on your calculation for CAP conduction velocity, how long would it take the CAP to travel the length of
the sciatic nerve? Assume a total length of 10 cm.
The conduction velocity of the action potential is determined by measuring the distance traveled (length of
the nerve in m) and dividing by the time (sec) taken to complete the reflex arc, also called the latency.
Conduction velocity = distance (m)/time (sec).
0.1 m / t = 155.4 m/s
t = 0.1 m / 155.4 m/s = 6.44x10
- 4
seconds


Exercise 3

Prelab Study Questions
1. Where is the gastrocnemius muscle and sciatic nerve in the frog and in you?
Gastrocnemius
largest and most superficial of calf muscles
part of Triceps Surae
main propellant in walking and running
Location in Human

Location in Frog

Sciatic Nerve
main nerve traveling down the leg
major branch of sacral plexus
innervates most of the hind limb
mixed-function nerve: made up of axons of sensory and motor neurons
Location in Human

Location in Frog



2. How does an action potential move in a bundle of neurons like the sciatic nerve?
compound action potential (recording from many axons) is a graded response
magnitude increases with intensity of stimulation
this is because different axons have different thresholds of excitation
largest axons have the lowest thresholds of excitation and are the most excitable
at low stimulus intensities, only the largest axons are activated
as stimulus intensity is raised in steps, more and more smaller axons are progressively recruited

3. What is the function of the PowerLab hardware and software?
Components of the PowerLab recording system.
Hardware
Transducer
Transforms the energy of a physiological event into electrical energy.
Amplifier
Amplifies the electrical signals from the transducer.
Data acquisition unit
Converts analog signals from the transducer or amplifier into digital information.
Computer
Records, displays, stores, and analyzes data.
Chart resembles a chart recording device.
Scope resembles an oscilloscope.
Software
LabTutor software
offers complete life science experiments, in one easy-to-follow web browser
interface including:
experiment background information
set up protocols
real-time data acquisition
laboratory reports

4. What type of signal does the Powerlab hardware receive?

5. What is tetanus?
infection of the nervous system with the potentially deadly bacteria Clostridium tetani (C. tetani)
infection begins when the spores enter the body through an injury or wound
often begins with mild spasms in the jaw muscles (lockjaw)
tetany: sudden, powerful, and painful contractions of muscle groups caused by prolonged muscular action;
may lead to muscle tears and fractures

Physiology of the amphibian skeletal muscle
Graded Response
Effect of Load on Contraction Force
Effect of Pulse Frequency on Contraction Force

Tetanus

Relationship between stimulus voltage and contraction strength
effect of stimulus strength
threshold stimulus, and maximal response at the lowest stimulus strength that results in some contraction
(i.e. Threshold), only a few motor units are stimulated
by increasing the strength of stimulus, an increased number of motor units can be "recruited" to increase
contraction force, and a greater displacement (muscle contraction) is recorded on the graph.

effect of stimulus frequency on skeletal contraction

muscle contraction
results in development of tension or force usually measured in grams
muscles will shorten if they develop more force than the force that is opposing them
ex: contracting muscles in our arm will shorten and allow us to lift a book if the force developed by the
muscles is greater than the weight (force) of the book.

no tetanus
stimuli are applied at a frequency where the time interval between stimuli is longer than the time it takes for
the muscle fibers to completely contract and relax, then no tetanus (no temporal summation) is observed

partial/incomplete tetanus
stimulus frequency was increased until the muscle fibers no longer had time to completely relax
partial relaxation between muscle twitches
most sustained voluntary skeletal muscle contractions are incomplete tetanic contractions with different
motor units stimulated at different times (asynchronous contractions)
asynchronous contractions delay muscle fatigue, which is an inability to contract caused by long periods of
muscle contraction
stimuli is applied at a frequency where the time interval between stimuli is shorter than the time it takes for
the muscle fibers to completely relax, then partial (incomplete) tetanus is observed
type of wave summation with partial relaxation observed between twitches


fused/complete tetanus
stimulus frequency was increased further so that the muscle fiber could not even begin to relax
sustained contraction with no relaxation observed between twitches
stimuli are applied at a frequency where the time interval is shorter than the time it takes for the muscle
fibers to even begin to relax, then fused (complete) tetanus is observed
type of wave summation with no observable relaxation between twitches

twitch contraction
type of muscle contraction, but not normal muscle contractions
single, brief stimulus is applied to a muscle fiber, either naturally in the form of a nerve impulse, or artificially
in the form of an electrical stimulus
quick shortening observed in a skeletal muscle when a single action potential traveling down a motor neuron
stimulates the skeletal muscle fibers of the motor unit to contract
single contractile event in response to single action potential


three phases of twitch contraction

latent period
lasts about 2 msec (milliseconds)
time between stimulation of muscle cells and force generation

contraction period
lasts about 10100 msec
period during which force (measured in grams) is increasing

relaxation period
which lasts 10100 msec
period when force is decreasing
phase of contraction; period during which more crossbridges detach than reattach to thin filaments

normal muscle contractions
not twitch contractions
sustained contractions of varying force

threshold stimulus
minimal stimulus that results in a muscle twitch

maximal stimulus
stimulus that produces maximal force is called the maximal stimulus
stimulus to the muscle greater than maximal does not produce a greater force

wave summation
if the muscle fibers of a motor unit are stimulated before the relaxation phase of a muscle twitch is complete,
then the next contraction will produce a greater force
how to increase force generated
increasing the frequency of muscle stimulation produces sustained force generation
increasing the number of motor units contracting at the same time, motor unit recruitment increases force
generated

maximal force development
occurs when all motor units of a muscle are stimulated and all muscle fibers are contracting

fatigue
an inability to contract caused by long periods of muscle contraction

wave/temporal summation
force of contraction seems to increase like the sum of the individual contraction waves

load
force that the muscle is contracting against.

displacement
strength of contraction
sum of the force exerted by all the motor units that are excited

recruitment
multiple motor unit summation
increasing the number of motor units that are stimulated to contract

motor unit
motor neuron and all the muscle fibers it innervate


Fatigue

Electromyography

Smooth muscle

Postlab Study Questions
1. In light of the all or none law of muscle contraction, how can you explain the graded response?
The frog sciatic nerve is made up of different axons, each with their own threshold. Some are higher than
others while some are lower. Therefore, as you increase the voltage, more and more of them fire, giving a
seemingly graded response
This graded response phenomenon illustrates the differences in threshold that exists among the different
sizes of fibers that make up the nerve. Remember, you are recording from a nerve, a large bundle of
neurons, each with a different threshold. If the stimulus voltage is increased slowly and smoothly, you may
observe discrete jumps in the amplitude of the compound action potential as different threshold classes of
nerve fibers are recruited. As you increase the amplitude more neurons reach their threshold and
contribute to the increase in size of the compound action potential. Eventually, as the stimulus voltage is
increased, a point will be reached when the wave form of the action potential stops changing. At this point all
the fibers in the nerve able to respond to the stimulus are being stimulated. This is a maximal response.

2. What effect does stretching the muscle have on contraction strength? Is this effect linear? What preload
force resulted in the highest contraction force?

3. What effect does varying the stimulation frequency have on contraction force? Which stimulus interval
caused the greatest contraction force?

4. Define tetanus. At which stimulus interval did you observe tetanus?
Definition
infection of the nervous system with the potentially deadly bacteria Clostridium tetani (C. tetani)
infection begins when the spores enter the body through an injury or wound
often begins with mild spasms in the jaw muscles (lockjaw)
tetany: sudden, powerful, and painful contractions of muscle groups caused by prolonged muscular
action; may lead to muscle tears and fractures
what stimulus interval
partial stimulus
stimulus frequency was increased until muscle had no time to completely relax
partial relaxation between muscle twitches
stimuli are applied at a frequency where the time interval between stimuli is shorter than
the time it takes for the muscle fibers to completely relax
fused/complete tetanus
stimulus frequency was increased further until muscle could not even begin to relax
sustained contraction wherein no relaxation observed between twitches
stimuli are applied at a frequency where the time interval is shorter than the time it takes
for the muscle fibers to even begin to relax

5. At what time point did your muscle begin to fatigue? Calculate the % decrease in contraction force by
comparing the force at the end of the experiment with the maximal contraction force.

6. In your own words, explain a possible mechanism for why the muscle was unable to maintain a prolonged
contraction in this experiment. .