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Physiology of the Nerve

Astrid Amador, Mark Joseph Amancio, Mariah Jemm Amurao, and Rex Augustus Artuz
4Biology4 Group 1
Department of Biological Sciences
College of Science
University of Santo Tomas

Abstract

The nervous system is a complex collection of nerves and specialized cells known
as neurons that transmit signals between different parts of the body. It is
essentially the bodys electrical wiring. Four activities were done to understand
Keywords:
the physiology of the nerve. Twitching was only observed through electrical
Nerve function, effect among others in testing the function of nerves. Applying anesthesia and
signal blocking, blocking the pressure increases the threshold voltage needed to elicit response
reflex action
as time goes by and had a minimum stimulus of 3 volts. The damaged brain and
spinal cord of the frog showed minimal to no response in comparison to the nonpithed frog. For the action reflex and reflex time, it was observed that the 10%
HCl gave the fastest reflex time averaging 0.56 sec.
Introduction
The nervous system consists of the brain, the
spinal cord and the nerves that conduct
impulses to and from the Central Nervous
System (CNS). The nervous system consists
of billions of neurons and neuroglial cells.
These neurons are able to be stimulated by
touch, light, sound and other stimuli, they
also conduct impulses and communicate with
each other. When the neurons receive
electrical or chemical stimulation they
produce an action potential. This study aims
to determine the effect of mechanical,
thermal, osmotic, and other factors on nerve
conduction, the effect of single and double
pithing on the different reflexes of a frog, to
determine the response of the nerves that are
exposed and not exposed to anesthetics, and
to test the time of reflex action of the frog in
different concentrations of acid.
Materials and Methods
Double-Pithing
The frog was grasped using the left hand and
the head was bended over the middle finger
with the index finger. The foramen magnum
was then located using dissection needle. The

dissection needle was pushed cranially into


the foramen magnum to destroy brain. The
dissection needle was then pushed caudally to
destroy the spinal cord.
Nerve Dissection
The skin from legs and abdomen of the
double-pithed frog was removed by cutting
around the abdomen and peeling the skin
downward and off the animal. The frog was
kept moist using Ringers solution. The
urostyle was grasped using forceps and was
cut. The sciatic nerve was then located and
lifted using a glass hook. The nerve from the
spinal cord was cut and was deflected back
onto the animals leg. A piece of thread was
tied around the free end of the nerve. The
nerve was severed from the gastrocnemius
muscle. The nerve was placed in a petri dish
containing frog Ringers solution.
A. Nerve Function
A muscle nerve preparation was made. The
nerve was stimulated by pinching
(mechanical), touching it with a hot glass rod
(thermal), applying a few crystals of NaCl to

the fresh cut (osmotic), dipping the free end


of the nerve in HCL (chemical), and applying
a mild electrical shock using stimulator
(electrical)
B. Signal Blocking
B1. Anesthetic solution
The minimum stimulus that will elicit a
response on the gastrocnemius muscle was
determined by touching one end of the nerve
with a stimulating electrode. Two cottons
with anesthetic were placed between the
middle portions of the nerve. This was
covered with filter paper moistened with
ringers solution. The free end of the nerve
was then stimulated using the minimal
stimulus previously determined. This was
repeated with two-minute interval and the
threshold stimulus was determined at each
stage. The cotton was moistened with
anesthesia periodically. The anesthetic was
washed away with amphibian ringers
solution and the minimum stimulus was
determined at two-minute intervals.
B2. Pressure Blocking
The minimum stimulus that will elicit a
response was determined. Pressure was
applied gradually using prongs of forceps
with parafilm on the middle portion of the
nerve, which was then released immediately.
The minimal stimulus was again determined
and the same procedure was repeated at 1minute intervals. The results were tabulated.
C. Functions of the Central Nervous
System
Nine physiological tests were performed for
a non-pithed, single-pithed and double-pithed
frog. It was first placed in a dissecting pan,
the position of the head, eyes and legs were
observed. The pulsation in the throat area was
noted. Next, in the dissecting pan, the animal
was placed on its back and the righting
position of the frog as the pan is revolved
horizontally was observed. The frog was then
placed in a pail of water and the pulsation in

the throat area was observed. The frog was


placed on the table and a loud noise was
made. The reaction of the frog was observed.
After that, the firmness and flaccidness of the
frogs legs was observed. Also, the hind leg
of the frog was pulled and was observed if a
resistance is present. Next, the toe of the frog
was pinched and its reaction was noted. Then,
the eyes of the frog were touched with a piece
of cotton or tissue paper. Lastly, the toes of
the right foot of the frog were soaked in
heated water in a beaker. All the procedures
were repeated and results were recorded.
D. Reflex Action in the Frog
The frog was single-pithed following the
pithing procedure. The frog was suspended
by the jaw with a wire hook attached to a
clamp on a stand. The long toe of the frog was
immersed with 10% sodium bicarbonate then
tap water and it was subsequently immersed
in 10% HCl. The time before the reflex action
obtained without the toe touching any part of
the beaker was noted. This procedure was
repeated after 2 minutes but using different
concentration of the acids.
Results and Discussions
Before examining the physiology of the
nerve, double-pithing procedure was done.
Double-pithing procedure is a known method
to immobilized amphibians for experimental
purposes. This is used to reduce the pain and
bleeding of the frog (Suckow, Murthy &
Silverman, 2007). After the double pithing,
brain was destroyed therefore perception of
sensory phenomena is impossible. Also,
spinal cord was destroyed which makes the
frogs legs limp. However, the frog remained
alive due to cutaneous respiration (Biopac,
2001).
After the pithing procedure, nerve dissection
procedure was performed. During this
procedure, the frog should be kept moist by
Ringers solution. Ringers solution is used to
create isotonic solution relative to bodily

fluids of animal. It is a solution that typically


contains sodium chloride, potassium
chloride, calcium chloride dissolved in water
(Adams, 2016). It is important to keep the
frog moist because the frog is still breathing
through the skin. After the nerve was
dissected, metal tools or fingers must not
touch it because this will cause the
depolarization of membrane, which can
affect future experiments. The nerve must
also be kept moist by ringers solution so the
nerve stays flexible. In addition, if it is dry,
the nerve becomes brittle and hence, will
break easily. (Biopac, 2001)
Table 2.1: Nerve Function

Stimulus
Mechanical
Thermal
Osmotic
Chemical
Electrical

Observation
No twitching
No twitching
No twitching
No twitching
Twitching
was observed

Expected
Twitching
of muscle

After dissection of the nerve, its function was


examined through mechanical, thermal,
osmotic, chemical and electrical stimulation.
Most of the stimulus given to the frog elicited
no response except for electrical stimuli as
opposed to the expected observation (Table
2.1). There should have been a response for
all the stimuli because frogs have sensory
receptors
i.e.
mechanoreceptors,
thermoreceptors,
nociceptors
and
photoreceptors that recognizes stimuli and
initiates sensory transduction. Sensory
transduction is a process that converts the
stimuli recognized by the sensory receptors to
neural signals so that an electrical response in
the cell membrane is elicited (McMillan,
2014).
Initially, a neuron is at its resting membrane
potential. i.e, the inside is negative relative to
the inside due to the presence of protein ions
with negative charge. In addition, during

resting membrane potential, most ion


channels are closed. However, some
potassium channels are open therefore
potassium can freely diffuse out of the cell.
The resting membrane potential of a neuron
is about -70 mV which means that the inside
of a neuron is 70 mV less than the outside.
When a nerve is stimulated, the sodium
channels are opened causing an influx of
sodium ions into the cell membrane making
the cell membrane more positive relative to
the outside. This is called the depolarization
state. When depolarization reaches about -55
mV, a neuron will fire an action potential
(threshold). If the neuron does not reach this
threshold level, no action potential is fired.
After a cell has been depolarized, sodium ion
channel begins to close. Meanwhile, the
sodium potassium pump also pumps three
sodium ions outside and two potassium ions
inside. This will re establish the resting
potential membrane of the nerve (Purves,
2001).
The probable reason behind the observed
result is that the threshold potential was not
reached creating no response and the nerve
was not kept moist.
Table 2.2: Signal and Pressure Blocking

Interval
1
2
3
4
5
6
7
8
9
10

w/

w/o

Anesthetic

Anesthetic

3
6
9
9
12
9
12
-

3
3
4.5
4.5
4.5
4.5
3
6
4.5
4.5

Pressure
Block
3
3
4.5
9
12
12
-

*Minimum Stimulus: 3 volts

After examining nerve function, the group


also examined the effects of blocking the

signals by applying anesthetic solution and


pressure.
Local
anesthesia
inhibits
depolarization of the nerve membrane by
interfering with Na+ and K+ currents. Based
on the results, applying anesthesia increases
the threshold voltage needed to elicit
response as time goes by. This is because the
sciatic nerve is composed of three types of
neurons based on diameter. Type A being the
largest, Type B moderate, and Type C, the
smallest. As a result, anesthesia block type C
fibers easily than type A fibers (Gymrek,
2015). In the early time interval, the effect of
anesthesia is not yet apparent because only
type C fibers are blocked. However, as the
time goes by, the Type A fibers are also
blocked therefore no response was elicited.
Meanwhile, when no anesthetic solution is
applied, the threshold voltage also increases
but very minimal compared to that with
anesthetic solution because the signals are not
blocked. However, the myelin sheath
degrades with time. As a result the impulses
are slowed down because the charges are not
insulated. On the other hand, when pressure
is applied, the nerve diameter was reduced
until such time that no more signals are able
to pass. In addition, the influence of pressure
in blocking conduction is entirely dependent
upon deformation of the tissue due to its
unbalanced action (Causey, 1949).
Table 2.3: Reaction of the single-pithed frog

Position of
the head
Righting
In Water
Reaction to
Noise
Firmness of
Legs
Eye
Response

Lying position, eyes


blinking, slow pulsation of
throat are
No reaction
No Reaction
No Reaction
Flaccid
Closes eye for a short
period of time

Table 2.4: Reaction of the double-pithed frog

Position of the
head
Righting
In Water
Reaction to
Noise
Firmness of
Legs
Eye Response

Lying position, eyes


are not blinking, slow
pulsation of throat
No reaction
Does not do its
righting position
No movement
Very flaccid
No reaction

Table 2.5: Reaction of the non-pithed frog

Position of
the head
Righting
In Water
Reaction to
Noise
Firmness of
Legs
Eye
Response

Normal position, blinking


eyes, fast rhythmic
pulsation of throat
The frog immediately
went to righting position
The frog immediately
went to righting position
The frog jumped away
from the source of the
noise
Firm hind limbs
Closes eye for a long
period of time

The non-pithed frog responded normally to


the physiological tests that were given while
the double-pithed frog has no reflex actions
at all since the brain and spinal cord are fully
damaged. However, the single-pithed frog
still has little response action in comparison
to the non-pithed frog due to the intact spinal
cord.
The CNS consists of the brain and the spinal
cord. Its main focus is to coordinate incoming
and outgoing neural impulses, integrating
data, and forming motor output. When CNS
receives a sensory input, it produces motor
responses through nerves. The central
nervous system is divided into the central,
peripheral, somatic and autonomic nervous

Table 2.6: Reflex Action of the frog in HCl

Conc.
2%
4%
6%
8%
10%

1st trial
2.88
1.09
1.00
0.61
0.43

Time (sec)
2nd trial
Mean
2.92
2.9
1.88
2.97
1.01
1.005
0.9
0.755
0.69
0.56

In this activity, we determined whether the


reaction time to a stimulus is influenced by
the strength of the stimulus. In this case, the
stimulus is the hydrochloric acid (HCl). The
skin of frogs and toads are more sensitive to
drought and chemicals in water because it can
absorb water and breathe through the skin as
blood vessels are found underneath it. Acidic
solutions can therefore be felt more by the
amphibian skin, especially in areas where
there is only a thin covering, like the toes,
which
is
also
convenient
for
determining the withdrawing reflex action
as it only exposes a minimal surface area, can
be easily dipped, and washed without
affecting most of the body. Using 10%
concentration HCl gave the fastest reflex

time, averaging 0.56 seconds, which is due to


the higher proton (H+) available of all the
concentrations. The acid disrupts the pH of
the frog skin demanding water and other
buffering agents from the skin as it is needed
to balance out the disruption. Therefore,
excess acid, like any other ions promotes a
feeling of dehydration (Doratt, 2008) to the
toad.
4
TIME!(SEC)

system. The central nervous system is made


up of the brain, spinal cord and nerves while
the peripheral nervous system consists of
sensory neurons, ganglia and nerves that
connect to one another and to the central
nervous system. Nerves in the peripheral
nervous system contain both sensory and
motor neurons. The somatic nervous system
is a division of the PNS that includes all of
the voluntary efferent neurons. The SNS is
the only consciously controlled part of the
PNS and is responsible for stimulating
skeletal muscles in the body while the
autonomic nervous system is a division of the
PNS that includes all of the involuntary
efferent neurons. The ANS controls
subconscious effectors such as visceral
muscle tissue, cardiac muscle tissue, and
glandular tissue

3
2
1
0
0%

5%

10%

15%

CONCENTRATION

Figure 2.1. Graph of the concentration of the acid


vs time in seconds showing that as the concentration
of the acid increases, the reaction time decreases.

The time 1.005 s and 0.755 s for 6% and 8%


respectively agrees that the lower the acid
concentration the less effect there is to the
frog giving longer reflex time. Exposure to
the acid is done from the lowest
concentration to the highest concentration
because the sense might get used to the acid
sensation in higher concentration and will
then tolerate the succeeding lower
concentration acids. Washing the frog with
10% sodium bicarbonate and water after
every exposure to acid is important to
stabilize and ideally get the previous
condition of the frog, and it is done together
with keeping the frog moist. Human errors
contributed might be not being able to wipe
properly the toe after washing with alkali and
water. Drying it too much might have
increased or could have diminished the
sensitivity of the toe compared to the natural
conditions or the time when the experiment
began when nothing has been done to the toe
prior to the experiment. The experiment

shows the susceptibility of the amphibian


skin to slight changes in acidity.
Conclusion
The observations say otherwise but stimuli of
different kinds (mechanical, thermal,
chemical, osmotic, and electrical) to the
nerve will elicit twitching of the muscle. In
signal blocking, by applying pressure to the
nerve, the diameter of the nerve becomes
smaller and there will come a time that the
signal will not be able to pass. Applying
anesthetic to the nerve increases the threshold
voltage needed to elicit a response. A nonpithed frog would react normally to
physiological tests, while a single pithed frog
will only react when its eyes is touched and
the eyes are still blinking. While a double
pithed frog would not react at all to the
physiological tests. The reflex action of the
frog in response to stimulus is faster in a high
pH condition.
References
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Connections to Nursing Practice (Second
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