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10 .Experiment

The experiment investigates the effects of physostigmine and atropine on the dose-response curve of acetylcholine in isolated frog rectus abdominis muscle. Physostigmine enhances muscle sensitivity to acetylcholine, shifting the curve left, while atropine blocks the response, resulting in a flattened curve. This study illustrates the pharmacological impacts of cholinergic agents on muscle contraction.

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shubham verma
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52 views3 pages

10 .Experiment

The experiment investigates the effects of physostigmine and atropine on the dose-response curve of acetylcholine in isolated frog rectus abdominis muscle. Physostigmine enhances muscle sensitivity to acetylcholine, shifting the curve left, while atropine blocks the response, resulting in a flattened curve. This study illustrates the pharmacological impacts of cholinergic agents on muscle contraction.

Uploaded by

shubham verma
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
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Download as DOCX, PDF, TXT or read online on Scribd

Experiment: Effect of Physostigmine and Atropine on the Dose-

Response Curve (DRC) of Acetylcholine Using Frog Rectus Muscle

Objective

To study the effects of physostigmine (a cholinesterase inhibitor) and


atropine (a muscarinic receptor antagonist) on the dose-response curve
(DRC) of acetylcholine (ACh) on isolated frog rectus abdominis muscle.

Materials

1. Frog rectus abdominis muscle preparation

2. Acetylcholine solution

3. Physostigmine solution

4. Atropine solution

5. Ringer’s solution (frog)

6. Isolated organ bath

7. Kymograph or data acquisition system

8. Syringe and pipettes

Procedure Overview

1. Isolate the rectus abdominis muscle of the frog and mount it in the
organ bath containing Ringer’s solution maintained at room
temperature.

2. Record the baseline contractions of the muscle induced by


increasing doses of acetylcholine (ACh).

3. Plot the initial dose-response curve of ACh.

4. Add physostigmine to the bath and record the muscle's response to


the same doses of ACh. Plot the modified DRC.

5. Wash the tissue thoroughly with Ringer’s solution.

6. Add atropine to the bath and record the muscle's response to the
same doses of ACh. Plot the modified DRC.

Observation Table
Dose of
Response (mm) Response (mm) with Response (mm)
ACh
without Drug Physostigmine with Atropine
(µg/mL)

0.1 2 5 0

0.2 4 8 0

0.4 6 12 0

0.8 8 16 0

1.0 10 18 0

Graph

 Plot 1: DRC of acetylcholine alone

 Plot 2: DRC of acetylcholine in the presence of physostigmine (Shift


left)

 Plot 3: DRC of acetylcholine in the presence of atropine


(Flattened/Blocked response)

Results and Discussion

1. Effect of Physostigmine:

o Physostigmine is a reversible cholinesterase inhibitor. It


prevents the breakdown of acetylcholine, leading to an
increased concentration of ACh at the receptor site.

o The dose-response curve shifts to the left, indicating


enhanced sensitivity of the muscle to ACh. Each dose elicits a
stronger contraction than before physostigmine addition.

2. Effect of Atropine:

o Atropine is a competitive antagonist of muscarinic receptors.

o The addition of atropine results in a flattened dose-response


curve. Even at higher doses of ACh, there is no response
because the muscarinic receptors are blocked, preventing ACh
from exerting its effect.

Significance
This experiment demonstrates the roles of cholinesterase inhibition and muscarinic
receptor blockade on acetylcholine-induced muscle contraction. It highlights the
pharmacological actions of cholinergic agonists and antagonists.

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