Understanding Sensory Transduction: Lab Report on Pain Detection

NAME: Robert McCants III (Rya Amour) DATE: November 18, 2023 Lab Report Sensory Transduction: Learn why you feel pain when you get hit by a rock This lab report is for you to reflect on what you completed and learned in this simulation, and to practice your written scientific communication skills. Include enough detail to clearly describe the lab experience to someone who understands the concepts but has not participated in the simulation. Use correct scientific terminology whenever applicable. Sections 1. Describe the overall objective and make a hypothesis 2. Introduce relevant background knowledge on this topic 3. Summarize the steps taken in the simulation 4. Explain any obtained results 5. Discuss the conclusions and implications 1 Copyright Labster ApS 2021 All Rights Reserved

1. Describe the overall objective and make a hypothesis. What is the overall purpose of the experiments or activities? Make a hypothesis. Recall that the hypothesis is a claim that answers a question or makes a prediction about an event that has not yet occurred. Hint: The purpose is often stated in the welcome message of the simulation (as shown in the example image below). For your entry make sure to include the following: ● Include one statement that describes the overall objective in your own words. ● Include one statement that describes the hypothesis. The primary aim of this experiment is to ascertain the specific medication that can alleviate pain without compromising an individual's ability to perform efficient muscle movements. 3 Copyright Labster ApS 2021 All Rights Reserved

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2. Introduce relevant background knowledge on this topic. What have you learned in class or researched on your own that would help prepare for this simulation? Hint: You can review the “THEORY” section in the simulation (as shown in the example image below) For your entry make sure to include the following: ● Include at least three important details from the lab “Theory” tab, the Lab Manual, or other resources, in your lab report that supports the simulation activity. Various types of stimuli are detected by distinct sensory neurons. In this particular study, the nociceptor is the most pertinent neuron as it responds to pain caused by mechanical, heat, or chemical stimulation. The nociceptor expresses TRPV1 channels on its surface, which alters the cell membrane's action potential from negative to positive. This change causes an influx of sodium into the cell, resulting in an action potential that triggers the sensation of pain. Capsaicin, a chemical, can directly impact TRPV1 channels. 4 Copyright Labster ApS 2021 All Rights Reserved

3. Summarize the steps taken in the simulation. Explain all steps you completed, including the equipment and techniques you used. Hint: You can use the “MISSION” tab in the LabPad as inspiration (as shown in the example image below). For your entry make sure to include the following: ● Document each step you completed, including the equipment and techniques you used. 1. The cell body of a nociceptor was isolated by taking a sample from the dorsal root ganglia of an animal specimen through dissection. This area is known to contain nociceptors, which are responsible for detecting pain. 2. To access the spinal cord, an initial incision was made using a scalpel. Scissors were then used to remove a portion of the spine, and forceps were used to separate a portion of the spinal cord from the spine. A portion of the dorsal root ganglia was then removed from the spinal cord and placed on a patch clamp plate for further study using a microscope. 3. By blocking calcium and potassium channels, the activity of sodium channels can be observed. 4. The ion concentration of the bathing solution surrounding the neuron, as well as the ion concentration inside of the neuron, can be measured using two electrodes. Additionally, the resting potential of the neuron's membrane can be changed with electrical current 6 Copyright Labster ApS 2021 All Rights Reserved

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using the electrodes. 5. The neuronal membrane potential experienced a minimal change when the resting membrane potential of -70mV was altered to -60mV. However, a significant alteration in the membrane potential was observed when the voltage was further modified to -5mV. 6. The inclusion of Lidocaine in the buffer solution, coupled with a voltage measurement of -5mV, demonstrated that Lidocaine effectively prevents the opening of sodium channels. Consequently, this inhibition of sodium channels should lead to a diminished perception of pain. 7. Subsequently, the buffer solution was modified to incorporate the drug Rebine while maintaining a voltage of -5mV. This experiment revealed that Rebine does not appear to have an impact on sodium channels, as the resulting graph closely resembled the control graph. Following this, the buffer solution was further modified to include capsaicin and Rebine, with the voltage remaining at -5mV. 8. Through the execution of a voltage test using the combination of Rebine and capsaicin in the buffer solution, it was determined that this combination does not inhibit action potential. It was observed that Rebine alone does not seem to activate sodium channels, and its effectiveness relies on the channels already being open. The Rebine/capsaicin combination allows for an increase in membrane voltage without triggering an action potential. 9. Subsequently, the effects of Lidocaine and Rebine were examined on a live animal specimen. 10. Following the injection of a control solution into the live specimen, the reflex time to heat-induced pain was measured and recorded. Subsequently, the specimen was injected with capsaicin and tested for reflex time to pain. The reflex time was slightly shorter, potentially indicating that the specimen was closer to its pain threshold due to the capsaicin injection. 11. Next, the specimen was injected with Lidocaine and tested for pain reflex time. The results indicated a slower reflex time, as Lidocaine inhibits the activation of nociceptors. When a mixture of Lidocaine and capsaicin was administered, a similar slow reflex time was observed, albeit marginally faster. 12. Subsequently, the specimen undergoes an injection of Rebine and undergoes a reflex test to assess its response to pain, yielding outcomes comparable to those observed with the control solution. Following this, a combination of Rebine and capsaicin is administered via injection, and the specimen's pain reflex is evaluated. The outcome reveals a marginally swifter reaction compared to the combination of Lidocaine and capsaicin. 13. The results of this examination indicate that Lidocaine has the ability to alleviate pain, regardless of the presence of capsaicin, whereas Rebine necessitates the presence of capsaicin in order to produce its pain-relieving effects. 7 Copyright Labster ApS 2021 All Rights Reserved

4. Describe any obtained results. Explain all obtained results. Were these results expected or unexpected? Hint: You can use the “MEDIA” tab in the Lab Pad to revisit relevant images from the simulation (as shown in the example image below). For your entry make sure to include the following: ● Document the results from the simulation. ● Document whether the results were expected or unexpected, and briefly explain why. When pain or capsaicin triggers the opening of TRPV1 channels, it allows the influx of positive ions. Consequently, Rebine is able to enter the cell more easily, which is otherwise hindered by the cell's bilayer membrane. In contrast, Lidocaine functions by blocking voltage gated sodium channels, thereby preventing their activation. Similarly, Rebine also inhibits these channels, but the manner in which they enter the neuron differs. Lidocaine is lipid soluble, enabling it to dissolve easily into transmitting cells such as motor neurons, thereby inhibiting action potentials. This can result in numbness and impaired muscle function, while simultaneously reducing pain. On the other hand, Rebine faces difficulty in entering cells due to its lack of lipid solubility. Consequently, it requires an open channel to facilitate its passage. This requirement aligns with the nature of a cell membrane's phospholipid bilayer. 8 Copyright Labster ApS 2021 All Rights Reserved

5. Discuss the conclusions and implications. How do your results relate back to the original purpose and your hypothesis? Were there any systematic sources of error that could have affected the results? What did you learn? For your entry make sure to include the following: ● Include a statement that briefly summarizes how the results of the simulation relate back to your original purpose and hypothesis. ● Include a statement that details any systematic sources of error you identified that could have affected the results. If the lab did not have any systematic sources of error, include a statement that acknowledges the lack of error present in the lab. ● Include a statement about what you learned from the simulation. Rebine exclusively interacts with nociceptors, and this interaction occurs only when TRPV1 channels are already activated, indicating the existence of pain. Thus, for Rebine to exhibit its efficacy, the presence of capsaicin is necessary. Consequently, unlike Lidocaine, Rebine does not impede the functioning of motor neurons. During my laboratory experiment, I acquired knowledge regarding the lipid solubility of Lidocaine, which renders it less specific in terms of its distribution within the body. On the other hand, Rebine can be employed with a more precise objective in mind. Additionally, prior to conducting this lab, I had not come across the drug Rebine. 9 Copyright Labster ApS 2021 All Rights Reserved

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Lab Report - Touchstone 4

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