BIOL2230 – Neuropharmacology Lab Report

Name:
Student
Number:

Lab report tasks/questions

Q1. Describe and explain the effects of nicotine on neural activity in the cricket TAG
preparation. (1) Using the PrtScn function, capture representative images of your
recordings displayed in the Audacity software at a size/resolution sufficient to
show the neural activity before and after injection. Label the traces to indicate the
moment of injection. (2) Use the RMS volume difference values (in dB) that you
calculated in Audacity to describe the changes in activity quantitatively. [3
marks]

Needle Out

Injection Point Response

The needle was inserted into the cricket at approximately 2.49s, as seen on the image, there is a slight spike
which indicates it. The nicotine was then injected and the needle was removed at 2.54s. The response is
shown at 3.00s where the neural activity was firing, causing the influx of spikes. The difference before vs
after RMS (dB) after injection is 2.28s and the difference after vs recovery RMS(dB) is 2.59s.

Q2. Explain why nicotine has the effect you observed. If what you saw did not
match your expectation suggest why [2 marks].

Nicotine is an antagonist for the neurotransmitter and acts as a stimulant which increases neural firing.
Nicotine acts as a stimulant to the CNS, causing a release of epinephrine which stimulates the nervous
system (Nicotine and Tobacco Products | Ochsner Health, n.d.). Injecting nicotine into crickets amplifies
their excitatory neurotransmitter, acetylcholine (ACh) (Effect of Nicotine and MSG on Neurons, n.d.). This
creates an increase in neural activity as nicotine amplifies the effect of ACh binding to its receptors in
synapses, causing a neuron to fire more due to increased sodium ion influx.

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 Q3. Describe and explain the effects of calcium chloride (CaCl2) on neural activity
in the cricket TAG preparation. (1) Using the PrtScn function, capture
representative images of your recordings displayed in the Audacity software at a
size/resolution sufficient to show the neural activity before and after injection.
Label the traces to indicate the moment of injection. (2) Use the RMS volume
difference values (in dB) that you calculated in Audacity to describe the changes
in activity quantitatively [3 marks].

Response

The needle was inserted into the cricket at approximately 4.40s, as seen on the image, there is a slight spike
which indicates it. The calcium chloride was then injected and the needle was removed at 4.49s. The
response is shown at 5.25 where the neural activity was firing, causing the influx of spikes. The difference
before vs after RMS (dB) after injection is 2.47s and the difference after vs recovery RMS(dB) is 0.91s.

Q4. Explain why CaCl2 has the effect you observed. If what you saw did not match
your expectation suggest why [2 marks].

Unlike nicotine, Calcium Chloride is an inhibitory since it stops sodium channels from opening. When the
Calcium Chloride was injected, it caused a decrease in the spiking of neural activity. This is due to the levels
of calcium ions being elevated, increasing both the voltage dependence on sodium and potassium ion
channels. (Hung et al, 2009). Blocking sodium channels reduces the velocity of action potential transmission
within the heart (Armstrong and Cota, 1999).

Q5. Describe and explain the effects of glutamate (L-glutamate/MSG) on neural

activity in the cricket TAG preparation. (1) Using the PrtScn function, capture
representative images of your recordings displayed in the Audacity software at a
size/resolution sufficient to show the neural activity before and after injection.
Label the traces to indicate the moment of injection. (2) Use the RMS volume
difference values (in dB) that you calculated in Audacity to describe the changes
in activity quantitatively [3 marks].

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The needle was inserted into the cricket at approximately 3.23, as seen on the image, there is a slight spike
which indicates it. The glutamate was then injected and the needle was removed at 3.29s. The response is
shown at 3.25s where the neural activity was firing, causing the influx of spikes. The difference before vs
after RMS (dB) after injection is 2.47s and the difference after vs recovery RMS(dB) is 0.91s.

Q6. (3) Explain why glutamate has the effect you observed. If what you saw did
not match your expectation suggest why [2 marks}.

When glutamate binds to the glutamate gated chloride receptors, it causes an influx of chloride anion into
the axon membrane, evoking a negative effect on the action potential firing. When glutamate is released, it is
stimulated by nerve impulses to the muscle. It then attaches to receptors which stimulate the muscle
contraction (Eichhorn, n.d.). As seen in the results above, the firing of neural activity decreased as glutamate
has a negative effect.

Q7. The most widely used class of modern insecticides, the neonicotinoids, mimic
the effects of nicotine on the insect nervous system. With reference to the
molecular action of nicotine can you explain why compounds that mimic the
effects of nicotine would be lethal to insects [3 marks].

Neonicotinoids are agonists of the insect nicotinic acetylcholine receptors (nAchRs), they are the main
excitatory neurotransmitters. When binding to nAchRs, neonicotinoids induce neural depolarisation (Cabirol
and Haase, 2019). Neonicotinoids are neurotoxins, insecticides and work by disrupting a protein receptor
located in the insect’s central nervous system. They inhibit the nervous system from working properly since
once it binds to the acetylcholine receptors, it blocks acetylcholine from binding which inhibits firing of
neutrons. This ultimately leads to the effect od paralysis and death (Beckwith, 2018).

Q8. In your experiments the difference in neural activity before and after injection is
considered to be the signal of the effect of your pharmacological compound on
the insect TAG. Suggest how you might have increased the signal in your
experiments [2 marks].

To increase the strength of the signal, everyone moved away from the cricket except for the person injecting.
This helped with excess noise, as there were too many movements and electrical noise which causes some of
the spikes in our results. So, it’s best to be in a quiet environment. Another way to increase signal could be
the usage of more electrodes on the points were there higher levels of neural activity.

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References:
Armstrong, C. and Cota, G., 1999. Calcium block of Na+ channels and its effect on closing rate.
[online] National Library of Medicine. Available at:
<https://www.ncbi.nlm.nih.gov/pmc/articles/PMC22436/> [Accessed 3 April 2022].
Backyardbrains.com. n.d. Effect of Nicotine and MSG on Neurons. [online] Available at:
<https://backyardbrains.com/experiments/teachersguide/neuropharmacology#:~:text=Nicoti
ne%20amplifies%20the%20effect%20of,to%20increased%20sodium%20ion%20influx).>
[Accessed 3 April 2022].
Beckwith, W., 2018. Neonicotinoid Pesticides Impair Bees’ Social Behavior. [online] American
Association for the Advancement of Science. Available at:
<https://www.aaas.org/news/neonicotinoid-pesticides-impair-bees-social-
behavior#:~:text=Neonicotinoid%20insecticides%20are%20neurotoxins%20and,death
%20of%20crop%2Ddestroying%20pests.> [Accessed 3 April 2022].
Cabirol, A. and Haase, A., 2019. The Neurophysiological Bases of the Impact of Neonicotinoid
Pesticides on the Behaviour of Honeybees. [online] Available at:
<https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6835655/#:~:text=Neonicotinoids%20are
%20agonists%20of%20the,acetylcholine%20%5B11%2C13%5D.> [Accessed 3 April
2022].
Eichhorn, L., n.d. Glutamate Receptors and the Righting Reflex in Crickets. [online]
Msacad.org. Available at: <https://msacad.org/journal/julyjournal/laura.html> [Accessed 3
April 2022].
Ochsner Health. n.d. Nicotine and Tobacco Products | Ochsner Health. [online] Available at:
<https://www.ochsner.org/health-resources/tobacco-free-living-the-legacy/tobacco-free-
resources/nicotine-and-tobacco-products#:~:text=Nicotine%20acts%20as%20both%20a,and
%2C%20over%20time%2C%20addiction.> [Accessed 3 April 2022].
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