Experiment 2.D and 2.

E: Function of the Central Nervous System and Reflex Action of the

Frog

NG, Phillip Matthew C.; NGO, Ricky Justin N.; Ramirez, Mary Adeline C.; REBOSA, Ariana;
REYES, Jojee

4BIO7-8

Department of Biological Sciences

College of Science, University of Santo Tomas
España, Manila, Philippines

Abstract I. Introduction

The nervous system is a network of nerve The nervous system is a complex

cells that transmit signals from different parts
of the body. There were five experiments to
understand the physiology of the nerve, two of
which were assigned to the group. Double-
pithing caused the animal to lose its capacity
for movement while single-pithing only
permitted involuntary movements performed
by the spinal cord. In examining the reflex
actions of the frog, it was also found that
reaction time and the strength of stimulus are
inversely related to each other.
network of nerves and cells that carry
messages throughout the various parts of the
body. It consists of the central nervous system
(CNS) and the peripheral nervous system
(PNS). The CNS composes the brain and the
spinal cord while the PNS includes the
somatic and autonomic nervous systems.
The basic unit of the nervous system is
the neuron. It is a specialized cell used to
conduct information electrochemically. The
three parts of a neuron are cell body, dendrites,
and axon. The cell body is the control center,
while the dendrites are branched extensions
which carry a nerve impulse into cell body and
the axon is a long, thin process that carries
away information from the cell body to
another neuron or tissue. Several neurons
bundled together are called a nerve.
Neurons are classified into sensory,
interneurons, and motor based on their
specific function. Sensory neurons conduct
impulses from sensory receptors to the central
nervous system. Interneurons conduct
information between neurons within the
nervous system. Motor neurons transmit
information from the central nervous system
to the effector systems.
 Neurons send information
electrochemically, meaning that a chemical
causes the generation of an electric signal.
Electrically charged ions cross through the
membrane of the neuron to produce an action
potential.
In this experiment, the students aimed
to determine the effect of single-pithing and
double-pithing on different reflexes of the frog
as well as the function of its central nervous
system.
II. Materials and Methods
A. Single-Pithing
In single-pithing a frog, it was held
with the left hand and its head was bent
ventrally over the middle finger. Using the
right hand, the dissection needle was then
inserted into a depression at the base of the
skull, which is the foramen magnum of the
animal. The needle was then pushed cranially,
destroying the brain.
B. Double-Pithing
In double-pithing, the same procedure
was done as with single-pithing. However,
after pushing the needle cranially and
destroying the brain, the dissection needle was
then directed caudally down the spinal column
to destroy the spinal cord, moving the needle
in a circular motion until the legs became stiff
and then completely flaccid as well as
unresponsive.
C. Functions of the Central Nervous
System
A non-pithed frog was first used and
placed on a dissecting pan. The position of its
head, eyes and legs, as well as the pulsation in
its throat area were observed. The frog was
then placed on its back and its righting
position was observed. The pulsation in the
frog’s throat was also observed when it was
put in a beaker of water. Afterwards, the frog
was placed on the table and its reaction to loud
noises was noted. The firmness or flaccidness
of the frog’s legs was also noted. The hind leg
of the frog was pulled gently to examine and
observe if any resistance was present. The
frog’s toe was then pinched and its response
was noted. The frog’s response to its eye being
touched by a piece of cotton was also
observed. Lastly, the toes of the frog’s right
foot were dipped in a beaker of hot water and
its reaction was noted. The same procedures
mentioned above were then repeated using
both a single-pithed and a double-pithed frog.
D. Reflex Action in the Frog
In this activity, a single-pithed frog
was used. The pithing procedure was done
according to the instructions given in the
manual. The frog was suspended by the jaw
using a wire hook affixed to a clamp on a
stand. The long toe of the frog was soaked
with 10% sodium bicarbonate, following with
tap water, and then subsequently immersed in
HCl. Without letting the toe touches any part
of the beaker, the time it took to obtain a reflex
action was noted. The same procedure was
repeated after two minutes, however, with
different concentrations of the acid in
increasing order.
III. Results and Discussion Retraction of
In heated
foot after some
water
A. Functions of the Central Nervous System time
In this experiment, the different
functions of the brain and spinal cord, both Results of the non-pithed frog are
being the main components of the central shown in table 3. Since both brain and spinal
nervous system, were tested by subjecting the cord were still functional in this frog, the
animal to a series of tests and taking note of its position of its head was in an upright position
responses to each. This was done to observe and when laid on its back, it immediately
which reactions are controlled by the brain and turned to the right to stand. This is the righting
which are controlled by the spinal cord, as reflex. When put in a beaker of water, the
well as which were still possible with only the pulsation in its throat was observed to be slow.
spinal cord left functional and if there were This is normal in frogs because when in water,
responses possible with both brain and spinal they keep their throat inflated to keep air in
cord no longer intact. A non-pithed frog, a their lungs so as to add buoyancy and to retain
single-pithed frog, and a double-pithed frog oxygen while in water. Its senses are also
were used. Generally, the non-pithed frog responsive since its brain was still intact, thus
responded normally to the different tests. The explaining its reaction to a loud noise,
single-pithed frog, with its brain disrupted and touching, pinching, and temperature. The
spinal cord still intact, had lesser responses in brain is primarily responsible for sensory
comparison to the normal frog. The double- information, balance, muscle control,
pithed frog, on the other hand, resulted in very breathing and eye movements. The spinal
little to no response at all since both its brain cord, on the other hand, connects the brain to
and spinal cord were destroyed. the different parts of the body, controls
Table 3. Responses observed on normal/non- skeletal muscle movements and responsible
pithed frog. for coordinating reflexes. It is for this reason
Observations that the frog was able to retract its foot when
Upright dipped in heated water and when there was
Position of resistance when its legs were pulled,
position/normal
the head considering that its spinal cord was still intact
position
Normal and therefore capable of reflex movements.
Righting
position
Table 4. Responses observed on single-pithed
Normal In water Slow pulsation
frog.
Reaction to Movement of Observations
noise head and body Leaning
Firmness Firm with Position of
Single- towards the
of legs resistance the head
pithed right
Eye Eyes Righting No response
response blinked/closed
In water No pulsation
 Reaction to Table 5. Responses observed on double-
No reaction
noise pithed frog.
Firmness of Flaccid; no Observations
legs resistance
Position of Head laid back
Slowly the head straight
Eye
closing of
response Righting No response
the eyelid
Pulsation in
When the same procedures for this In water throat is less
experiment were repeated using a single- forceful/slower
pithed frog, results were different from the Left arm
Reaction to
normal frog. Results of the reactions observed Double- twitched; eyes
noise
from the single-pithed frog are shown in table pithed closed
4. In a single-pithed frog, the brain was Flaccid; no
destroyed but the spinal cord was still intact. resistance; toe
Firmness of
Destroying the brain disrupts the processing of did not
legs
sensory information as well since, as respond to
mentioned above, the brain is responsible for pinching
interpreting information from the senses. This Eye Eyes closed
is why the frog did not react to the loud noise response and retracted
and pinching. However, when its eye was In heated
No reaction
touched, it slowly closed. This could probably water
be due to the reason that some nerve endings
were not completely blocked yet. A part of the For the last part of this activity, the
brain also controls the righting reflexes of a same procedures were done on a double-
frog, specifically the rhomboid sinus and pithed frog. Results observed on the double-
medulla oblongata, which is why there was no pithed frog are shown in table 5. In a double-
reaction from the frog when it was laid on its pithed frog, both brain and spinal cord were
back. Disrupting the brain has resulted to the destroyed. In comparison to the single-pithed
loss in righting reflex. Since the brain is also frog, its left arm twitched as a reaction to the
responsible for breathing, specifically loud noise and its eyes retracted as a reaction
controlled by the autonomic nervous system, to corneal reflex. It is expected that the
no pulsation was observed in the frog’s throat animal’s perception to senses was already lost,
when it was placed in water. In addition to however, such results may have been possibly
this, there was also a loss in voluntary caused by an error in pithing or by reflex
movements, which explains why there was no muscle spasms as a response to the pithing.
resistance when the legs of the frog were Spasms normally occur when the muscle
pulled and they were already flaccid. receives nerve impulses to contract. Aside
from this, a much slower pulsation, in
comparison to the non-pithed frog, was also
observed when the frog was placed in water.
Despite the destruction of the brain, this
occurred possibly because pithing may not
have been done properly, as mentioned above,
or that there was still air left in the lungs. As
for the other tests done, the frog had no
reactions since double-pithing subjected the
animal to paralysis and muscle/reflex
movement was lost.
D. Reflex Action in the Frog
The central nervous system is in
charge of coordinating sensory information
and generating a motor output accordingly.
One of its components is the spinal cord,
which conveys messages between the brain
and the rest of the body, being the body’s main
“information highway”. Aside from
transmitting messages, the spinal cord is
responsible for reflex actions. According to
Raven and colleagues (2014), a reflex is an
immediate, involuntary muscular movement.
The rapid motor response to a stimulus is
made possible as the sensory input bypasses
the brain and travels along the reflex arc found
in the spinal cord. Instead, the sensory neuron
sends the information to a motor neuron along
the spinal cord without undergoing advanced
stages of processing and association. Hence,
in this activity, a single-pithed or decerebrated
frog was used to delimit the neural impulses
within the route of the spinal cord.
Table 6. Response time obtained from each
acid concentration used
Acid Concentration
Time (Seconds)
(%HCl)
2 3.45
4 1.95
6 1.33
8 1.25
10 0.75
In this activity, it was ascertained
whether the strength of the stimulus influences
the reaction time to such. Hydrochloric acid
(HCl) was used to serve as the stimulus. It was
observed that the highest concentration of acid
generated the fastest response while the lowest
concentration produced the slowest reaction.
Figure 1. Linear graph showing the
relationship between reaction time and acid
concentration
Figure 1 shows that the relationship
between acid concentration and reaction time
was expressed through a negative-sloped line,
indicating that the two variables are inversely
related. The data obtained from the
experiment coincided with the trend observed,
suggesting that the stronger the stimulus, the
faster is the reflex action. This can be
explained by the effect of stimulus intensity to
action potential. When the intensity of the
stimulus is strengthened, it does not bring
about increase in the amplitude of the action
 potential. It, instead, increases the frequency
of the action potential, leading to the opening
of more sodium channels, which causes the
influx of greater sodium ions, resulting to a
faster depolarization. Alternatively, at low
intensities, the stimulus can only cause few
channels to open, reducing the frequency of
the action potential and in turn generating a
slower response.
The experiment was carried out in the
order of increasing acid concentration to
prevent the frog from tolerating and being
desensitized to the lower concentrations.
Furthermore, rinsing the frog with 10%
sodium bicarbonate and water following each
exposure to the acid is vital to balance out the
effects of the acid and maintain the condition
of the frog in a steady state.
II. Conclusion
At the end of the experiment, it can be
concluded that double-pithing destroys both
the brain and spinal cord, causing paralysis.
As for the influence of stimulus intensity to
reaction time during a reflex action, it can be
inferred that reaction time is inversely
proportional to the strength of the stimulus.
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