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PhysioEX: Nerve Impulse

S.J.A. Orense, L.K. Palma, C. Pineda, E.V. Ragodon, K.D. Reyes
Summary
Nerve impulse is a wave of action potentials moving along the nerve fiber. It is the propagation of action
potentials along a nerve fiber. An action potential is a sequence of depolarization and repolarization. An
action potential at the trigger zone causes an electric current to flow a short distance down the nerve
fiber, which stimulates the next membrane to its threshold level, triggering another action potential. In
this experiment, electrical stimulation, mechanical stimulation, thermal stimulation and chemical
stimulation were done unto the sciatic nerve. In the electrical stimulation, the minimum voltage that
elicited an action potential was at 3.0V while the maximum voltage that elicited an action potential was
at 5.0V. In mechanical stimulation, the glass rod was used and the nerve elicited an action potential was
conducted, in thermal stimulation the nerve also responded from the heated glass rod and in chemical
stimulation; both HCl and NaCl elicited an action potential. Lidocaine and curare are effective in inhibiting
action potentials.
Introduction
Various agents affect nerve transmission and
different kinds of stimuli trigger an action
potential. Unequal distribution of ions in the
cells inner and outer membranes polarizes a
nerve cell. A polarized membrane is stimulated
at its threshold or above its threshold. When a
membrane overcomes its threshold, a
physiological effect begins to be produced. This
wave constitutes a nerve impulse, and if it
reaches a muscle, the muscle may respond by
contracting (Marieb, 125).
In this experiment a segment of a nerve has
been dissected and suspended over a series of
metal bars and reagents that act as electrodes.
The electrodes are stimulating electrodes and
are connected to an electrical stimulator. The
stimulator can be used to apply electrical current
to the nerve at different voltages and
frequencies to try to elicit an action potential.
The other set of electrodes are called recording
electrodes, and they are connected to an
oscilloscope. Differences in charge between the
two recording electrodes cause the line traced
on the oscilloscope screen to deflect. Thus, we
can observe any action potentials forming in the
nerve (Marieb 130-131). Effects of HCl, NaCl, and
heat and glass rod were also taken into
consideration in generating nerve impulse.
While in inhibiting production of an action
potential, lidocaine, curare and ether were used.
The objectives of this experiment are to test
and identify different stimuli capable of
generating an action potential and identify
stimuli capable of inhibiting an action potential.
Materials and Methodology
The experiment was conducted using a PhysioEx
5.0 software and a Laptop computer.
Electrical Stimulation
Upon opening the program of PhysioEx, the main


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menu was clicked and the Neurophysiology and
Nerve Impulses was selected. The voltage was
set to 1.0V by clicking the (+) next to the Voltage
display and the Single Stimulus was clicked. If
there was no response, the clear button was
selected and the voltage was increased together
with the clicking of the Single Stimulus button,
this step is being repeated until a deflection of
the line is seen which indicates an action
potential, this is termed the Threshold Potential.
The Data was recorded by clicking the Record
Data button. If printing is necessary, the tools tab
was selected and then Print Graph is clicked. The
voltage was increased by 0.5V and Single
Stimulus is clicked. The deflection line is
compared to the previous line and was recorded
by clicking the Record Data button. The voltage
was progressively increased by 0.5 V and the
Single Stimulus button is clicked until the point
beyond which no further increase occurs in the
peak of the action potential is found, this is
referred to as the Maximal voltage. The data was
recorded.
Mechanical Stimulation
Using the same opened program of PhysioEx, the
clear button was clicked. The glass rod located
on the bottom shelf on the left side of the screen
was dragged over to the nerve. Then the glass
rod is over the nerve, the mouse button was
released to indicate that the rod is not touching
the nerve. Then the oscilloscope screen was
observed whether a tracing has occurred or not.
The data was recorded by clicking the Record
Data.
Thermal Stimulation
Using the same opened program of PhysioEx, the
glass rod was clicked and was dragged on to the
heater. The Heat button was clicked and when
the rod turns red, this indicates that it has been
heated already. The heated-rod was clicked and
dragged over the nerve and the oscilloscope was
observed.
Chemical Stimulation
The dropper from the bottle of sodium chloride
was dragged over to the nerve in the chamber
and the mouse button was released to dispense
the drops. The effect was observed. The
threshold voltage setting from the Electrical
stimulation was compared. The data was
recorded by clicking the Record Data button. The
clean button was clicked and together with the
clear button. The dropper from the bottle of
hydrochloric acid was dragged over to the nerve
and the mouse button was released to dispense
the drops. The effect was then observed and was
compared to the original threshold stimulus. The
data was recorded by clicking the Record data
button. The clean button was clicked. If wished
to print the data, tools tab was selected and the
Print data button was selected.
Testing the Effects of Ether
The dropper from the bottle of ether was clicked
and dragged over to the nerve in between the
stimulating electrodes and recording electrodes.
The mouse button was released to dispense the
drops. Using the threshold voltage setting from
Electrical stimulation, the Stimulate button was
clicked the result was observed. The result was
recorded by clicking the Record Data button. The
time button on the oscilloscope was clicked until
the screen shows a display activity over the
course of 10 minutes. The (+) button under the
Interval between Stimuli on the stimulator was
clicked until the timer is set for 2.0 minutes. The
Stimulate button was clicked and the Elapsed
Time display is observed. When the nerve
returns to normal, the Stop button was clicked to
stop the action and return to the Elapsed time to
0.0. The Time (msec) button was clicked to
return it to its normal millisecond display. The
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clear button was clicked and the (-) button under
the Interval between Stimuli was clicked until it
is reset to 0.00.
Testing the Effects of Curare
The dropper from the bottle of curare was
clicked and dragged on to the position of the
nerve in between the stimulating and recording
electrodes. The mouse button was released to
dispense the drops. The stimulator was set at the
threshold voltage and the nerve was stimulated.
The effect on the action potential is noted. The
data was recorded by clicking the Record Data
button. The clean button was selected and the
clear button was clicked to clear the oscilloscope
screen.
Testing the Effects of Lidocaine
The dropper from the bottle of lidocaine was
clicked and dragged on to the position over the
nerve between the stimulating and recording
electrodes. The mouse button was released to
dispense the drops. The tracing were observed.
The data was recorded and if printing is
preferred, the Tools tab was selected and Print
Data was clicked. The clean button was clicked to
remove the lidocaine and return the nerve to its
original untouched state.
Measuring Nerve Conduction Velocity
The Pulse button was clicked and the bio-
amplifier was turned on by clicking the
horizontal bar on the bio-amplifier and dragging
it to the On setting. The dropper from the bottle
of ethanol was clicked and dragged over the
earthworm and the mouse button was released
to dispense the drops of ethanol. The earthworm
was clicked and dragged into the nerve chamber,
using the (+) button next to the Voltage display,
the voltage was set to 1.0 V then the stimulate
button was clicked. The action potential was
observed. The Measure button was clicked and a
vertical yellow line appears on the far left edge
of the oscilloscope screen. The (+) button was
selected which moved the yellow line to the
right. The time elapsed was measured on the
graph by positioning the yellow line at the point
in the graph where the graph ceases being a flat
line and first starts to rise. The data were
recorded. The earthworm was clicked and
dragged to its original place and the clear button
was clicked. The other test organisms were
tested using the same procedures and the data
were recorded. The data were printed by clicking
the tools tab and the Print data button was
selected.
Results and Discussion
I. Eliciting a Nerve Impulse
Neurons are cells that correspond by
transmitting stimuli to other cells. They have two
major physiological properties: irritability and
conductivity. Irritability is that capability of
neurons to respond to electrical impulses
converting them to nerve impulses. On the other
hand, conductivity is the neurons capacity to
transmit nerve impulses.
In the experiment, a sciatic nerve dissected
from a frog was used. The stimulus travelled
from the red and black leads: from the stimulator
output to the nerve chamber and also to the
oscilloscope. The current produced by the
oscilloscope travels to the nerve, the nerve then
depolarizes and the electrical current that
develops flows continuously to the leads.
Starting the experiment, the voltage was set to
1.0V in order to see if an action potential would
occur. At 1.0V, there was no action potential that
took place. The voltage was increased and the
minimum voltage that elicited an action
potential was at 3.0V. Once the threshold was
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established, the voltage was again increased to
observe difference in peaks of the action
potentials. A slight increase in height of the
peaks of the different action potentials are
observed until no difference was seen even
though there was still a continued increase in the
voltage. The maximum voltage was at 5.0V.


Graph 1.1 Increasing action potentials corresponding
changes in the voltage

Figure 1.1 Comparison of different voltages whether it will
elicit an action potential
The next activity in this experiment was to
determine whether the nerve would respond to
mechanical, thermal and chemical stimulus. In
the mechanical stimulation, a glass bar was
placed on the nerve to see if it will draw out a
response from the nerve. By placing the glass bar
on the nerve, you will observe that there was an
action potential coming from the nerve
displayed on the oscilloscope screen. The tracing
displayed was lower than that of the threshold
voltage which was at 3.0V. For thermal
stimulation, the glass bar was heated and placed
on the nerve. It produced a higher peak on the
oscilloscope screen than the unheated glass bar.
This means that the heated glass rod conducted
a higher action potential than the unheated glass
bar.


Graph 1.2 Comparison of the action potentials elicited in
mechanical and thermal stimulation.
Red line indicates the heated glass bar while the green line
displays the tracings for the unheated glass bar.



Figure 1.2 Comparison on the mechanical and thermal
stimulation.

Chemical stimulation was also performed in
the experiment. NaCl and HCl were added to the
nerve to observe if certain action potentials
would occur. Based on the Figure 1.3, we can
detect that both chemicals triggered an action
potential; also, both tracings were the no
different than the tracings displayed by the
threshold voltage.

Figure 1.3 Comaprison of Nacl and HCl in maxing out an
action potential.

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Conclusion
Neurons respond to different stimulus may it be
electrical stimulation in where you apply electric
currents (voltage) in triggering a response;
mechanical stimulation in which you elicit a
response by use of glass bars; thermal
stimulation in where you apply heat in certain
objects and placing it on the nerve; and chemical
stimulation in where you add different chemicals
like NaCl and HCl on the nerve.
II. Inhibiting a Nerve Impulse
Table 1. Summary of Results for PhysioEx experiment on
Nerve Impulse Inhibition






As several factors could elicit a nerve impulse, a
myriad of external causes may also lead to
inhibiting action potential. For example,
chemicals, particularly local anesthetics do so by
inhibiting the depolarization of the nerve
membrane through interference of both Na
+
and
K
+
currents. The action potential is not
propagated because the threshold level is never
attained (Gmyrek et al., 2013). In this
experiment, three chemical agents, were tested
to inhibit a nerve impulse.

Lidocaine

Lidocaine is a local anesthetic drug whose
inhibiting process is both transitory and
reversible and is chiefly serving purposes in
dental surgery, ventricular heart arrhythmias,
and gastroeneterologic matters.
Figure 1. Screen shot of normal nerve impulse at
minimum voltage
Figure 2. Summary screenshots of PhysioEx results for
Nerve Impulse inhibition using Lidocaine, Curare, and
Ether with Curare only showing positive action
potential
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This anesthetic works by easily binding and
blocking the fast voltage gated sodium (Na+)
channels to be found both on the heart
cardiomyocytes and in the neuronal cell
membrane, held responsible for signal
propagation. given that it alters signal
conduction in neurons. With sufficient blockade,
the membrane of the postsynaptic neuron will
not depolarize and so fail to transmit an action
potential, leading to its anesthetic effects (Vono,
2011).
In the simulation, this inhibitory effect was
observed when Lidocaine was dropped on the
muscle. No action potential was recorded at
minimum voltage as well as when the voltage
was doubled and reached maximum.





Curare


Curare is a skeletal-muscle
relaxant drug belonging to the alkaloid family of
organic compounds (Encyclopaedia
Britannica). It was used before for arrow poisons
to cause paralysis on the target.
Curares mechanism involves competing with
acetylcholine (which is responsible in stimulating
muscles) in binding with neuromuscular
junctions thus preventing muscle contraction.
This however only occurs postsynaptic and the
release of acetylcholine which is happening
presynaptic continues, recognized as an action
potential and is then still detected and recorded
in the simulation as shown in Figure 4.





Figure 3. Screen shot of Lidocaine trials showing negative
action potential despite application of increasing voltage
Figure 4. Screen shot of Curare trial showing positive
action potential
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Ether

Ether is an example of general anesthetic. This
type of drug decrease brain activity by opening
K+ gates; thus allowing these ions out of the cell.
The neuron becomes hyperpolarized as in the
absolute refractory period, and is unable to
discharge. Hyperpolarization prevents the
neuron from receiving another stimulus thus no
action potential is achieved (Charand, n.d) as
shown in figure 5.
Conclusion
Inhibition of a nerve impulse may occur due to
cancellation of depolarization as exhibited in
local anesthetics or through hyperpolarization as
effected by general anesthetics. Moreover,
processes occurring postsynaptic do not affect
the release of action potential.
References
Charand, K.X. (n.d.). Action Potentials. Retrieved
from
http://hyperphysics.phyastr.gsu.edu/hbase/
biology/actpot.html.
Gymrek, R et al. (2013, Jun 3). Local and
Regional Anesthesia. Retrieved from
http://emedicine.medscape.com/article/183
1870-overview.
Marieb, Elaine N., and Susan J. Mitchell.
"Neurophysiology of Nerve Impules:
Computer Stimulatiom." Human Anantomy
& Physiology. N.p.: n.p., n.d. 125-33. Print.
Vono, M. (2011, Jan 17). Lidocaine. Retrieved
from
http://flipper.diff.org/app/items/info/3239.
Encyclopaedia Britannica (n.d.) Curare.
Retrieved from
http://global.britannica.com/EBchecked/topi
c/146779/curare.
(n.d.) Outside Electrode with Action Potential.
Retrieved from
http://psych.athabascau.ca/html/Psych289/
Biotutorials/2/out_act.shtml.

Figure 5. Screen shot of Ether trials showing negative
action potential despite application of increasing
voltage