Experiment 3: Frog Nerve

Carpio, Simbillo, Sharma, Torio

Introduction
Neuron
● Nerve cell
● Building block of nervous system
● Transmits information
● 4 Major structures: dendrites, cell
body, axon, synaptic terminal
● Nerve consists of a bundle of axons
2 Types of Electrical Signals in Neurons
1. Graded Potentials
● Local membrane change
● Excitatory or Inhibitory
● Occurs in dendrites and cell bodies
● Signals vary in amplitude depending on the strength of stimulus
● Can summate in 2 ways:

Spatial Summation and Temporal Summation

2. Action Potential

● All-or-none response does not deteriorate

● Generated by sodium-potassium pump
● Travels long distances
Generation of Action Potential
How does action potential travel?
● Action potential moves down the
axon until it reached synapse.
● Wave of depolarization opens
voltage-activated Ca2+ channels.
● Influx of Ca2+ cause vesicle to fuse
with presynaptic membrane.
● Transmitter diffuses across
synaptic cleft and binds to
receptors on post-synaptic cell.
Compound Action Potential
● Algebraic sum of all individual fiber action potentials of the nerve
Refractory Periods
● Period of time during which neuron cannot generate another action potential.
1. Absolute Refractory Period
● Even an stronger normal stimulus will not being another AP
● Inactivated Na+ (must return to resting state)

2. Relative Refractory period

● Suprathreshold stimulus will be able to start an AP

● Potassium channels are still open, but Sodium channels have closed
Objectives
● Describe the nature of CAP
● Explain how increasing stimulus strength results in increased magnitude of CAP
up to some maximum
● Know how absolute value and relative refractory periods of a nerve can be
demonstrated and understand the physiological difference of these periods
● Determine the nerve conduction velocity
Methodology
Connect red & black alligator
clips from stimulator
electrodes → two of the
metal rungs on opposite side
of the Nerve Bath
- red BNC connector:
positive (+) analog
- black BNC connector:
negative (-) analog
output connector.
Threshold Voltage & Maximal CAP Amplitude
Refractory Period
Conduction Velocity
Results & Discussion

Figure 1. Relationship between stimulus

and response in frog sciatic nerve
Stimulus Voltage vs. Response Voltage
● Stimulus voltage and response voltage are directly proportional with each other
● Response voltage is the Compound Action Potential (CAP) of the nerve
● CAP is the algebraic sum of all action potentials of individual nerve fibers
● Increased stimulus voltage results in increased CAP
Results & Discussion

Figure 2. Relationship between stimulus

interval and amplitude of 2nd CAP in frog
sciatic nerve
Results & Discussion

Table 1. Conduction velocity of the frog sciatic nerve

Formula for Conduction Velocity:
Conduction Velocity
● Conduction velocity is the speed at which the electrochemical impulse travels
along a neural pathway
● Factors that influence conduction velocity:
- Diameter of axon
- Larger diameter means less leakage of of ions
- Less leakage of ions results in faster conductance
- Myelin Sheath
- Acts as an electrical insulator
- Action potential travels from one node to another via saltatory conduction
Conclusion
● The Compound Action Potential (CAP) is the algebraic sum of all the individual
fiber action potentials of the nerve.
● Increase in stimulus strength = more fibers are recruited resulting to more action
potentials adding up = producing a wider CAP with longer duration.
● The threshold voltage is the voltage needed in generating at least one action
potential from a fiber in a sciatic nerve bundle; thresholds are much lower.
● When the A-alpha fibers that make up the nerve have been excited and are
conducting CAPs, the size of the CAP will no longer increase.
Guide Questions:
1. How does a CAP differ from a single action potential?
Compound action potential is the summed action potentials for many
individual nerve cells, while a single action potential comes from a single
neuron or a single muscle fiber.
1. Action potentials are said to be all or none responses. Why does the frog sciatic
nerve give a graded response?
- The frog’s sciatic nerve gives a graded response because the nerve is a bundle
of axons. Therefore, when one axon is stimulated, it generates a smaller
response and if all axons are stimulated at once, it generates a larger
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?

The smallest voltage required to produce the maximum CAP was 400 mV. In
order to produce maximal response, all nerves must be excited.
4.In this exercise, you examined the effect of increasing stimulus intensity on the
nerve. What other stimulus parameter might also affect the nerve's tendency to
generate a CAP?

Other stimulus parameter that might affect the nerve’s tendency to generate
a CAP is the stimulus frequency.
5.Explain the difference between the relative and absolute refractory periods.

- Relative refractory period is the interval immediately following the absolute

refractory period, where the initiation of a second action potential is
inhibited but not impossible. While an absolute refractory period is when the
second action potential cannot be stimulated at all.
6. Briefly describe the cellular events responsible for the refractory period. (Hint:
Discuss the mechanism of repolarization).

- The cellular event responsible for the refractory period is the build-up of
potassium during repolarization because during its transition to
hyperpolarization, the build-up of potassium makes it more difficult for the
stimulus to reach the threshold.
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.

- Based on our calculation for CAP conduction velocity, it would take 0.0013
seconds for the CAP to travel 10 centimeters of sciatic nerve.
References:
Brink Jr, F. Linear range of Na+ pump in
sciatic nerve of frog. AJP – Cell
Physiology. 1983; Vol 244, Issue 3 198-C204

Boundless. (n.d.). Boundless Anatomy and Physiology. Retrieved from

https://courses.lumenlearning.com/boundless-ap/chapter/nerves/

P.(n.d.). Refractory Periods. Retrieved from

https://www.physiologyweb.com/lecture_notes/neuronal_action_potential/neuronal_action_potential_refractory_periods.ht
ml

Characteristics of the CAP. (n.d.). Retrieved from http://www.medicine.mcgill.ca/physio/vlab/other_exps/CAP/character.htm

Effects of nerve impulses on threshold of frog sciatic nerve fibres. (n.d.). doi:10.3897/bdj.4.e7720.figure2f