C.

CHEMICAL BLOCKING OF NERVE CONDUCTION

RESULTS

Table 2.3. Contraction of gastrocnemius muscle in reaction to anaesthesia.

GROUP ANAESTHESIA CONDITION TIME (min)

1 Present Anaesthesia Reached 10

Absent Recovery Time 2

2 Present Anaesthesia Reached 10

Absent Recovery Time 4

3 Present Anaesthesia Reached 10

Absent Recovery Time 4

4 Present Anaesthesia Reached 10

Absent Recovery Time 2

Blocking the nerve conduction is widely used in basic experimental neuroscience

such as reflex, autonomic, motor pathways blocking. In addition to that, it is also
widely used in applied neurophysiology and neuroengineering research which dwells
on the therapy development for the pain and spasticity. It is has been mentioned that
there are four methods on how to achieve nerve conduction block and one of those is
through chemical means (Ackermann et al., 2011).

In chemical blocking, the methods include the use of denaturing agents. This has
the ability to induce a permanent nerve block. This is due to the irreversible axonal
damage that the chemicals have brought. It specifically targets the channel block
(Ackermann et al., 2011).
 The chemical used in the laboratory experiment is the chloroform. It is classified
as a general anaesthetic. Looking at its chemical structure, a halogen is added to the
hydrocarbon backbone. Aside from the increase in its potency, the flammability is
heightened as well. Chloroform is known in the war times as a very potent anaesthetic
that has an appreciable analgesic and neuromuscular activity. Chloroform is
administered through inhalation and it is a good muscle relaxant. One of its downsides
is its carcinogenic effect, classifying chloroform as both hepatotoxic and nephrotoxic.
It also causes severe adverse circulatory effects such as arrythmias and hypotension.
Based on its unaccepted therapeutic index, chloroform is now banned to use in
surgeries as an anaesthetic (Kar, 2005).

The said anaesthetic prevent action potential in all excitable membranes to

generate and propagate. Once the unionized anesthesia diffuses inside the cell
membrane, anesthesia particles block the channels from the inside. Since sodium ions
cannot enter the cell, action potential cannot happen and it now becomes impossible
for it to reach the threshold which is -55mV. Action potential is vital and must be
reached for the activation of the neuromuscular junction.

The nerve terminal is concerned with the storage, synthesis, mobilization, release,
and recycling of the neurotransmitter. Once the action potential has reached the
presynaptic terminal of a motor neuron, it then activates the voltage-dependent
calcium channels. It is now open to accommodate calcium ions inside the neuron.
These calcium ions bind to the sensor proteins that are found in the synaptic vesicles.
Because of this, the release of the vesicle fusion with the cell membrane and
neurotransmitter, which is acetylcholine for vertebrates, from the neuron to the
synaptic cleft is triggered. In the synaptic cleft, there is a rapid breaking down of
acetylcholine by the enzyme acetylcholinesterase. After this process, choline is
transported back into the axon terminal. The choline is now recycled to make more
acetylcholine.
 Using general laws of physics, one can state that cells that are small in diameter
are more sensitive than with larger diameter. This is the primary reason why the
conduction in nerve fibers are blocked first than the muscle fibers. In an anaesthetic, it
abolishes the conduction of the fiber by conveying the pain sensation first. It blocks
the nerve conductance in both motor and sensory neurons. Since nerve conduction is
blocked, pain sensation is lost. The motor functions are impaired also (Kar, 2005).

Table 2.3 shows the time it took for the following conditions, anaesthesia reached
and recovery time, to take place. All groups follow the same trend; it took around 10
minutes for the chloroform to reach anaesthesia time. And it took 2-4 minutes to
recover from the effect of anaesthesia. Both are qualitatively measured based on the
contraction of the muscle, gastrocnemius, through the stimulation of the sciatic nerve.
Basically, this part of the lab experiment aims to understand the principle behind
anesthesia, which is the neuromuscular junction.

For the neuromuscular transmission to recover, two processes are involved. In the
first process, recovery happens when the anesthesia is eliminated from the
neuromuscular junction. While in the second process, an increase in the acetylcholine
in the neuromuscular junctions has an effect on the anticholinesterase (Hemmings and
Egan, 2013). In the laboratory experiment, the Ringers solution is used to wash away
the anesthesia. It belongs to the first process of neuromuscular transmission recovery
wherein the anesthesia is eliminated. The Amphibian Ringers Solution (ARS)
mimics the isotonic body fluids of the amphibians and it is composed of NaCl, KCl,
and CaCl2. Since it has available ions of sodium, it can somehow induce action
potential to the neuromuscular junction. This is the reason why after 2-4 minutes, the
muscle recovered from the anesthesia and it is evident when the muscle contracted.

Guide Questions
3. Discuss the importance of the neuromuscular junction and the mechanism involved.
What is the role of acetylcholine?

Neuromuscular junction is a synapse of the motor neuron and muscle fiber. In

between the synaptic end bulbs of the motor neuron and the motor end plate of the
muscle fiber, this synapse occurs. This motor end plate functions to initiate action
potentials across the muscle surface thereby causing muscle contraction. In addition,
the motor end plate is the highly excitable region of the muscle fiber of the plasma
membrane (Guyton and Hall, 2006).

The main importance of the neuromuscular junction is its ability to transmit

signals to the muscle fiber in order to cause muscle contraction. Muscles need to have
innervation in order to perform its function. It needs to avoid atrophy so it needs to
maintain its muscle tone (Guyton and Hall, 2006).

An action potential that reached the presynaptic terminal of a motor neuron

activates the voltage-dependent calcium channels. This in turn, allows the entry of
calcium ions in the neuron. The calcium ions bind to the synaptotagmin or the sensor
proteins found in synaptic vesicles. This triggers the release of the vesicle fusion with
the cell membrane and a neurotransmitter (acetylcholine for vertebrates) from the
neuron to the synaptic cleft (Guyton and Hall, 2006).

Acetylcholine is a known neurotransmitter that has a role in the diffusion across

the synaptic cleft. It binds to the nicotinic acetylcholine receptors or nAChRs on the
cell membrane of sarcolemma, a muscle fiber. Once the acetylcholine binds to the
receptor, it can depolarize the muscle fiber. After that, a cascade results and
eventually allows muscle contraction to happen (Levitan and Kaczmarek, 2015).

Literature Cited
Ackermann, D., Foldes, E. L., Bhadra, N., Kilgore, K. L. 2011. Nerve conduction
block using combined thermoelectric cooling and high frequency electrical
stimulation. HHS Public Access, p. 2.

Guyton, A. C. & Hall, J. E. 2006. Textbook of Medical Physiology (11th ed.).

Philadelphia, Pennsylvania: Elsevier Saunders Incorporated, pp. 85-87.

Hemmings, H. C. & Egan, T. D. 2013. Pharmacology and Physiology for Anaesthesia:

Foundations and Clinical Application: Expert Consult. Elsevier Health
Sciences, pp 272-285.

Kar, A. 2005. Medicinal Chemistry. Pharmacology. New Age International, pp.

75-79.

Levitan, I. & Kaczmarek, L. 2015. Intercellular Communication. The Neuron: Cell

and Molecular Biology (4th ed.). New York, NY: Oxford University Press, p.
153.