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Physiology laboratory
Exercises 1-4 and PhysioEx 1-4
Exercise 1: Cell Water Potential
Water Potential
 Tendency of water to move
into or within a system (animal
or plant tissue)
 Water moves from an area
that has high water potential
to an area with low water
potential
 The potential energy in water
as well as the difference
between the potential in a
given water sample and pure
water.
Two factors that affect water
potential:
1. Pressure
2. Amount of solute
Equation:
Ψsystem = Ψtotal = Ψs + Ψp + Ψm.
Ψs: solute or osmotic potential;
always a negative sign
Ψp: pressure potential; turgor
pressure; it may be -, 0 or +
Ψm : matric potential; due to water-
binding colloids in the cell; negative
in sign
 Solute potential (Ψs)
decreases with increasing
solute concentration; a
decrease in Ψs causes a
decrease in the total water
potential.
Determination of Water Potential
 When a tissue is placed in a
sugar solution, there is a net
movement of water
 Water readily moves across
the cell membrane through
protein-lined channels
 If the concentration of both
sides of the cell membrane
are not equal, there will be a
movement of water
molecules
1. Hypertonic
 Movement of water out of the
tissue
 Concentration of WATER is
greater INSIDE the cell
 Concentration of dissolved
solutes are greater outside
the cell
Animal cell (no cell wall):
Shrink or shrivel
Plant cell (with cell wall):
Plasmolysis
2. Hypotonic
 Movement of water into the
tissue
 Concentration of dissolved
solutes are less outside the
cell than inside
 Concentration of WATER is
more outside the cell
Animal cell (no cell wall):
Will burst
Plant cell (with cell wall):
Turgor pressure
3. Isotonic
 Equilibrium
 No net movement
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 Total molar concentration of
the dissolved solutes are
equal
 A 0.9% solution of NaCl
(saline) is isotonic to animal
cells.
 No effect on animal and
plant cells
Determination of solute potential
of extracted sap by cryoscopy
Cryoscopy
1. Examination of liquids based
on the principle that freezing
points vary according to the
amount and nature of the
solute
2. OSMOTIC POTENTIAL of a
solution (Based on osmotic
pressure and solute
concentration)
 The freezing point of an
aqueous solution decreases
with an increase in dissolved
solutes.
Equation:
Calculating for the solute potential:
Ψs = 1.22Tf
Ψs = solute potential in MPa at 0
Celsius
Tf= Freezing point of solution in
Celsius
Solute Potential of extracted sap at
room temperature
Ψs = 1.22Tc
c= the correction factor or room
temperature in K/s73K
Determination of water potential
of the plant sample
 The water potential of the
plant sample is the tendency
of the water to move in and
out of the plant sample
 Computed value
 Plant samples have cell walls
Exercise 2: Physiology of the
Nerve
The Nervous System
 Made up of the Central
Nervous system (Brain and
Spinal cord) and the
Peripheral Nervous System
 The main component or part
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of the nervous system is the
neuron or the nerve cell Three Phases of the Action
Parts of the Nervous Potential:
System 1. Resting
1. Cell Body 2. Action
2. Axon 3. Recovery
- Nerve impulses away
from the cell body (to the The resting potential is defined as
different parts of the body) the potential difference across the
membrane of the axon when there is
3. Dendrites no impulse is conducted; the action
- Nerve impulses toward potential is when an electric shock,
the cell body pH change, or a mechanical situation
 Dendrites and Axons are stimulates a nerve. The action
covered by the myelin sheath potential brings about nerve
 Secreted by the Schwann stimulation or a change in potential.
cells Thus in this phase, as the stimulus is
applied, an increase in the graph is
Types of Neurons shown. Depolarization occurs, as the
1. Affarent Neuron or Sensory inside of the axon is more positive at
Neuron this level. The recovery phase is
2. Efferent Neuron or Motor between the transmissions as K+
Neuron ions are returned to the inside and
3. Interneuron the Na+ ions are returned to the
outside, actively. It comes after the
The Action Potential repolarization of the action potential.

 The action potential is

generated by the opening and
subsequent inactivation of
voltage-gated sodium
channels and, with a slight
delay, the opening of voltage-
gated K channels.
 When inactivated, Na ions will
not be able to enter.
 These channels will continue
to be deactivated for a
specific period of time. Thus,
during that brief time of
inactivation, action potentials
cannot pass through. This
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period is called the absolute
refractory period.
 K channels help return the
voltage to the resting potential
after the action potential has
passed
 When restored, Na will be
able to go in.
 The period in which a stronger
stimulus must be applied in
order to produce an action
potential is called the relative
refectory period.
A. Nerve Function
 For every stimulus introduced
there is a response. (The
greater the stimulus, the
greater the response)
 Increase in temperature
indicates an enhancement of
the twitch-force as muscles
are activated at higher
temperature, and also events
that occur at the
neuromuscular junction by an
action potential of the nerve
cell, are all sensitive to
temperature.
 More Cl- atoms, more
contraction occurs. (The
presence of chloride that
prevents the muscle from just
contracting and instead
respond only to conscious
control or spinal reflexes,
thus it is only when there is a
high amount of that there
is a response. The addition of
NaCl and HCl also show that
as more ions, the more
the muscle will contract in
response to conscious
contraction of the muscles.
*seen in osmotic and
chemical stimuli)
 Higher or greater voltage,
greater response. The
voltage represents the
neurotransmitters.
(Example: acetylcholine)
B. Signal Blocking
 Application of anesthetic and
pressure blocks the signal of
stimulus generating a SLOW
response or NO response.
 Anesthetic: The anesthetic
applied affects the secretion
of the neurotransmitter
acetylcholine!
Acetylcholine is found at
the neuromuscular
junction of the muscle that
causes it to contract.
 The anesthetic has an
INHIBITORY response.
 Antibiotic used in the
experiment: Lidocaine
 The application of pressure in
the middle causes the signal
to be blocked.
C. Functions of the Central
Nervous System
 The interaction between the
primary parts of the central
nervous system, the brain
and the spinal cord, and their
effects to the physiology of
the body is the main focus of
this experiment.
 Decrease in the
responsiveness as more
of the central nervous
system was destroyed.
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 There still remains little
movement due to the
spinal reflexes of the frog
left in the spinal cord even
after destroying the brain.
D. Reflex Action in the Frog
 The reflex indicated in this
example is the stretch
reflex. It is a stretch reflex
or a myotatic, knee-jerk
reflex.
 As the impulse is sent to
the spinal cord via the
sciatic nerve and
immediately responds to
contract the muscle.
E. Measurement of Compound
Action Potential
 The CAP is the summation
of all the axons firing.
However, the CAP has a
limit since there is only a
limited amount of axons per
neuron.
Determination of threshold
voltage and maximum Compound
Action Potential
 As seen in Figure 1, the CAP
amplitude failed to rise above
0mV when the stimulus
amplitude is less than 50mV.
 It was only when 50mV was
applied that the CAP amplitude
started to rise.
 The CAP amplitude continued to
rise as the stimulus amplitude
was increased.
 The maximum CAP amplitude
was acquired at 3.350mV when
170mV of stimulus was applied.
When the maximum CAP
amplitude was acquired,
increasing the stimulus amplitude
no longer went over the
maximum CAP amplitude value.
4
CAP Amplitude (mV)
3
2
1
0
0 200 400 600
-1
Stimulus Amplitude (mV)
Figure 1. CAP amplitude versus
stimulus intensity
 Nerves are governed by the all-
or-nothing law wherein there is
no median action potential. A
stimulus will either activate the
nerve or it will not. As seen in
figure 1, the nerve only began
conducting action potentials above
50mV. More and more axons were
activated as the strength of the
stimulus increased.
 However, at 170 mV, all of the
axons within the nerve were
already activated. Thus CAP did
not rise above 3.350mV even if
the stimulus amplitude was
increased. Thus, the threshold
voltage of the nerve is 50mV while
the maximum CAP is 3.350mV.
Determination of refractory period
Interval Stimulus Second Action
Between Voltage Potential?
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Stimuli (mV)  The inactivation of the Na
(msec) voltage-gated channels is
250 20 Yes observable when double pulses
125 20 Yes are applied to a nerve. As the
60 20 No interval is decreased, a stimulus
60 25 No higher than the threshold voltage
60 30 Yes
is needed to produce a second
30 30 No
action potential. A higher stimulus
30 35 No
is needed since most of the Na
30 40 No
30 45 Yes
channels are still recovering from
15 60 Yes the previous depolarization. The
7.5 60 Yes absolute refractory period was
3.75 60 No determined by applied the
maximum amount of stimulus
 The threshold voltage of the voltage at a 3.75msec interval.
nerve used in PhysioEx 9.0 was The absolute refractory period of
set at 20mV. With an interval of the examined nerve is 3.75msec.
250msec and 125msec, a second The nerve needs more than
action potential was produced. 3.75msec for the Na channels to
Threshold voltage: Starting return to a resting potential. The
voltage (rising from 0) relative refractory period
 However, when the interval was immediately follows the absolute
lowered to 60msec, a second refractory period. In the relative
action potential was not refractory period, a second action
observed. It was only when the potential can be produced at a
stimulus voltage was raised to higher stimulus.
30mV that a second action
potential was observed. When
the interval was further
decreased to 30msec, it needed
45mV stimulus to generate a
second potential.
 To determine the maximum Determination of nerve
refractory period of the nerve, the conduction velocity
maximum amount of stimulus,
60mV, was applied. A 60mV Axon Myeli- Distance Time Conduction
stimulus generated a second Type nation from R1 between Velocity
to R2 APs (m/sec)
potential when the intervals were (m) (sec)
lowered to 15msec and 7.5msec. A
However, a 60mV stimulus failed Heavy 0.1 0.002 50
fiber
to produce a second potential at B
Light 0.1 0.01 10
a 3.75msec interval. fiber
C None 0.1 0.1 1
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fiber
 A fiber was heavily myelinated
and had a large diameter.
 B fiber had a medium diameter
and was lightly myelinated.
 C fiber was thin and
unmyelinated.
 A fiber needed the least time to
travel 0.1 meters with a velocity
of 50m/sec while C fiber took the
longest in conduction and has the
slowest conduction velocity of
1m/sec.
 The conduction velocity is
heavily dependent on the
amount of myelination and
diameter of the axon.
 Since myelin sheaths lack
voltage gated channels, they
function by salutatory conduction,
allowing the action potentials to
occur at only unmyelinated nodes
of Ranvier. Action potentials
‘jump’ when passing through
myelin sheaths and generate a
potential only on the nodes of
Ranvier in between sheaths. This
process further accelerates the
movement of electric signals.
Somatic Senses
Tactile Localization
Part of Body Localization
Difference
Arm 20 mm, 15 mm
Palm 8 mm, 4 mm
Over the palm 21 mm, 5 mm
Neck 15 mm, 4 mm
Leg 16 mm, 4 mm
Cheek 10 mm, 3 mm
Forehead 5 mm, 5 mm
Nose 10 mm, 3 mm
 Tactile localization is the
stimulus perception at a specific
part of the body (Crosskey,
Elithorn & Piercy, 1953). In this
part of the experiment, the result
shows that all first trial has
higher localization difference
than the second one. This is
possibly because precision of
locating the origin of the stimulus
is connected to the stimulus
intensity and also the receptor
density where the stimulus is
located. Also, the receptors in the
dermis are known to be not
distributed evenly, hence, the
different parts of the body such
as the arm and neck, can have
different ability to ascertain a
particular stimulus (Cliffors,
Harris & Karloy, 2006).
 Localization is greater the first
time the stimulus is generated
Touch Receptors Adaptation
No. of Coins Time of
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Perception
1 42.29 s
2 5.49 s
3 14.12 s
 As seen in table 3, the
number of coins that have the
highest time of perception is 1
with 42.29 seconds and 3 coins
being the lowest time of
perception, 5.49 seconds. The
results show phenomenon called
sensory adaptation. Since most
sense receptors strongly respond
to sensitive changes in its
surroundings, it stops reacting
when a particular stimuli become
steady or constant (Lumpkin,
Nelson, & Marshall, 2010). For
instance, when a person enters a
room he/she quickly adapts to the
odor of the room or touch
receptors stops in giving
information of our clothing until
change in stimuli.
Temperature Receptors
previous sensory adaptation
(Paxinos, 2012).
 Moreover, a process when an
area of the body receives painful
stimulus but the other area is the
one that perceived it is called
referred pain (Evans-Martin &
Cooley, 2004). In the results,
hands soak in the ice water felt a
sense of numbness which
eventually becomes a stinging
sensation while for the hands in
the 45°C water it stings at first but
stop. The feeling of pain was
send to the other area.
Hearing
Watch tick test for auditory acuity
Distanc Distanc Differen
e from e ce
ear to where
point subject
where can
subject hear
cannot again
hear

Water Sensation
 Since the left hand was Temperature
placed in iced water and the right Ice Water Sense of
hand in water at 45°C, the numbness
strength of stimulus slowly which
decreases until both temperature eventually
receptors have adjusted to their develops into a
new environment. stinging
 Change in baseline of the sensation
receptors can be observed if both 45°C At first, there
hands were place in average was a stinging
temperature water. sensation which
 This shows that temperature is eventually stops
thus not absolute but relative Righ 140 120 70
to the baseline created by the t
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Left 185 110 75
 The watch tick test was done to
analyze whether an individual
has the inability to hear high
pitched sounds. The subject
was observed to stop hearing
the ticking watch from the right
ear at the 140cm mark and was
able to hear it again at the
120cm mark. From the left ear,
the subject stopped detecting
the sound at 185cm and was
able to hear it again at 110. The
differences of the results from
the right and left ear were 70
and 75 respectively. The
closeness of the differences
can indicate that the subject
does not have loss of hearing
high frequency sounds.
Considered as a bedside
hearing test, the watch tick test
does not provide accurate
information on what type of
hearing damage a certain
patient has developed. This
method however is a useful test
of acuity that could allow an
individual to assess himself if
he has problems in hearing
(Dillon 2006).
Auditory adaptation
Sensation
Normal Same sound for
both ears;
Scramble sound
Rubber Tubing Louder sound
Pressed
 The test in auditory adaptation
is done to observe the process
in which changes in the
sensitivity of sensory
receptors occur in relation to
the stimulus. The subject in the
experiment was observed to
hear a louder sound after the
release of the pressure in one
of the tubes of the stethoscope.
The ears adapt to loud
sounds as it hits the bones
located in the inner ear.
 This makes the inner bones
contract and causes the
reduction of transmission of
sound vibrations to the inner
ear. (Nevid 2014)
 In the case in the experiment,
the subject heard a louder
sound on the ear where the
tube was pressed than the
other ear where the sound went
without any obstruction. It can
be deduced that the ear
connected to the tube without
any obstruction had adapted to
the loud sound. This results to
the bones in the inner ear to
contract reducing the sound
transmission. The other ear
however, did not have the
bones in the inner ear contract
due to the prevention of sound
to reach the ear. The release of
the pressure then gave the
impression that one ear is
hearing louder than the other
because of different
contractions on each ear.
Vision
Binocular Vision and Space
Perception
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The coin was initially seen at the middle of
the view during the beginning of the
experiment. When the left eye was closed,
the coin could only be seen at the right
portion and at the corner of the view. When
the right eye was closed the coin could be
viewed at the left portion of the whole view.
 The test in binocular vision
and space perception was to
observe the concept of having
two eyes producing two different
images but only processing one
image in the brain. The observer
in the experiment sees the coin in
two different places depending on
which eye is closed and sees it
as one object in one place when
both eyes are open. Sensory
correspondence explains that
binocular vision is the unification
of visual excitations from
corresponding retinal images into
a single visual image. An object
localized in one and the same
visual direction by stimulation of
the two retinas will appear as one.
Meaning the stimulus to sensory
fusion is the excitation of
corresponding elements. (Von
Noorden 1996).
Visual Acuity
As the size of the letter decreases in the
chart, the edge of each letter also gets
blurry. Specific letter look similar to each
other like letter E and F.
 The visual acuity test is used
to determine the smallest letters
one can read on the Snellen’s
chart. It is commonly used as a
routine part of an eye
examination. Visual acuity is
expressed as a fraction. The
numerator corresponds to the
distance between the chart and
the observer (usually 20 feet).
The denominator indicates the
distance at which a person with
normal eyesight could read the
same line you correctly read. The
subjects in the experiment stated
that as the size of the letter gets
smaller, the edges of each letter
also gets blurry. Mistakes in a
line on one or two letters can still
indicate that the subject still has
vision equal to that line. (Tasman
& Jaeger 2012).
Exercise 4: Physiology of Skeletal
Muscles
 The basis of locomotion is
dependent to the overall
physiological principles of the
skeletal muscles.
 Its activities are controlled
voluntarily, but regulated
subconsciously, wherein the
stimulus is supplied by a nerve
impulse.
 Skeletal muscles are made up of
long bundles of muscle fibers,
which are large, elongated and
multinucleate cells. Muscle fibers
are comprised of sarcomeres,
which are contractile units,
responsible for the relaxation and
contraction of the muscles.
 The mechanism of muscular
function is initiated by the action
potential that is elicited from the
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contact of the motor neuron and Continuous Stimulus
the muscle fiber at a common  With increasing stimulus
point called the neuromuscular amplitude, there are plenty of
junction. This interaction triggers conditions that could be observed
the release of acetylcholine, depending on the intervals of the
which diffuses across the stimulus.
sarcolemma and eventually binds  It could be unfused or fused
to the motor end plate, causing tetanus. Unfused or incomplete
the shift in the permeability of tetanus is produced when partial
ions and the graded response of relaxation is evident.
the muscle.  Fused or complete tetanus is
 A twitch is defined as a muscle produced, when partial relaxation
contraction responding to a single is not present.
and rapid stimulus from an
isolated action potential induced
by muscle fibers.
 Muscle fatigue is a condition
that the physical performance of
muscles can temporarily decline
due to a wide range of
physiological impairments such
as the generation of insufficient
motor command and the
accumulation of specific
metabolites in myofibers.
 Multiple contractions of a muscle
produce a summation of all the
tensions that is significantly
higher than the value of a single
twitch. This summation is termed
as the tetanic contraction or
tetanus.

Pulse Stimulus
 Increase in stimulus amplitude
(V)
 An increase in V leads to a an
increase in number of motor units
thus the contraction increases.
 This shows a positive relationship
between the contraction force
and the stimulus amplitude