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Physiology & Pharmacology Practical Preparation Exercise 2, Schemes and Mind Maps of Biological Sciences

University of LeicesterBiological Sciences

A detailed overview of a computer-based 'dry' practical exercise on understanding antagonists using the guinea pig ileum experimental setup. It includes a description of the experimental setup, concentration-response data for acetylcholine in the presence of different concentrations of the antagonist atropine, and a guide on the schild plot analysis. The exercise aims to reinforce the principles of receptor pharmacology, drug-receptor interactions, and the use of drugs to define receptors and investigate physiological processes. Students are expected to construct, analyze, and interpret concentration-response curves, schild plots, and understand key pharmacological terms. The document also outlines the assessments for the practical component of the module, which includes a multiple-choice quiz and a laboratory report. Overall, this document serves as a comprehensive preparation resource for students to develop their understanding of antagonist pharmacology and data analysis skills.

Typology: Schemes and Mind Maps

2023/2024

Uploaded on 05/05/2024

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BS2013: Physiology & Pharmacology
Practical Preparation Exercise 2
2022-2023
Practical preparation: a computer-based ‘dry’ practical.
Exercise 2: understanding antagonists.
This document contains a brief description of the Guinea pig ileum experimental set up used to
generate the data you will be plotting, concentration-response data collected with different
concentrations of antagonist present, a guide on the Schild plot, and questions for you to
answer in exercise 2a (i) – (vi) and exercise 2b (i) – (vi).
This exercise can be completed after watching the Panopto video called ‘Practical preparation
exercise 2’. Aim to complete exercise 2 by the Q and A session so you can bring any queries.
Please post any questions ahead of time on the discussion board so we can collate your queries
and answer frequently asked questions in the live session. You can also ask questions live in
the Q + A session.
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BS2013: Physiology & Pharmacology

Practical Preparation Exercise 2

Practical preparation: a computer-based ‘dry’ practical. Exercise 2: understanding antagonists. This document contains a brief description of the Guinea pig ileum experimental set up used to generate the data you will be plotting, concentration-response data collected with different concentrations of antagonist present, a guide on the Schild plot, and questions for you to answer in exercise 2a (i) – (vi) and exercise 2b (i) – (vi). This exercise can be completed after watching the Panopto video called ‘Practical preparation exercise 2’. Aim to complete exercise 2 by the Q and A session so you can bring any queries. Please post any questions ahead of time on the discussion board so we can collate your queries and answer frequently asked questions in the live session. You can also ask questions live in the Q + A session.

Assessments Assessment of the practical component forms 40% of the module marks. The MCQ assessment based on the practical preparation material (‘dry practical’) is worth 10 %. This will be given on Friday 2 8 th^ October. Although it is a multiple choice style, please keep in mind that you will have to plot graphs, analyse and interpret data in order to answer the questions. It is estimated that it will take you approximately 3 hour to complete so allow plenty of time. The practical report, based on the laboratory or computer simulation of the experiments on Guinea pig ileum (‘wet practical’) is worth 30% of the module marks. This laboratory report write up is estimated to take you approximately 4 hours. Exercise 1 and 2 will help prepare you for both assessments.

Experimental set-up

A piece of ileum (lumen flushed to clear gut contents) is threaded at either end, mounted in an organ bath and set under tension/load of 0.5 g (this should make the preparation fully relaxed). The tissue is bathed at 37oC in physiological solution (Ringer’s) bubbled with carbogen (a mix of 95% oxygen and 5% carbon dioxide). The effects of drugs (agonists and antagonists) applied to the organ bath on ileum contraction can be measured in this system. In addition, responses to nerve stimulation can also be determined. Nerves can be stimulated electrically by field stimulating electrodes in the organ bath. Short duration electrical pulses (< 1 ms) can be used to selectively activate nerves. Typical trace showing the contractile responses of the Guinea-pig ileum with 100nM ACh with wash off and return to baseline and then an addition of 300nM ACh with wash off.

Exercise 2 a: Atropine antagonism of acetylcholine responses The aim of this exercise is to look at the action of the antagonist atropine. You have been provided with the results from the concentration-response to ACh from exercise 1 but this time in the presence of 3 separate concentrations of atropine, 0.001M (1E-9M), 0.03M (3E-8M) and 1M (1E- 6M). Plot these data using Excel. Amplitude of response ACh concentration (M) No atropine control Atropine 1 X10-^9 M Atropine 3 X10-^8 M Atropine 1 X10-^6 M 1X10-^09 0 0 0 3X10-^09 0 0 0 1X10-^08 10 2 0 3X10-^08 29 16 0 1X10-^07 46 33 1 3X10-^07 54 48 10 1X10-^06 56 56 30 3X10-^06 60 59 46 1X10-^05 58 60 54 3X10-^05 60 60 57 1X10-^04 60 60 59 3X10-^04 60 60 60 1X10-^03 60 60 60 3X10-^03 59 60 60 1X 10 -^02 60 60 60 3X10-^02 60 60 60 Answer the following questions. (i) What is the effect of atropine on the concentration-response to ACh? (ii) From your answer what type of antagonist is atropine? (iii) What does this high concentration of atropine do to the EC 50 of ACh? (iv) How experimentally could you discriminate between a reversible competitive antagonist and a low concentration of an irreversible competitive antagonist? (v) From your results what receptor is ACh acting upon? (vi) Create a diagram to outline the signalling steps in the ileum muscle whereby ACh evokes a contractile response.