Practical work.........83 31. Electrical stimulation on leg muscle of frog Procedure 1. Kill a frog with double pith method. 2. Cut open the abdominal wall of the frog. 3. Nerve-muscle preparation: • Identify the sciatic nerve and tie it with a cotton thread (Fig.1). • Trace along the sciatic nerve to the leg muscle. • Tie Achilles tendon of leg muscle with another cotton Fig.1 Identifying the sciatic nerve thread. • Separate the leg muscle from the sciatic nerve bones and neighbouring tissues by cutting the connective tissue and Achilles tendon. 4. Place the nerve-muscle preparation in Ringer’s solution completely until in use (Fig.2). 5. Use a pin to hold the muscle at knee joint at one side. 6. Fig.2 Nerve -muscle preparation Apply an electric current with a low voltage of about 2 V to stimulate the electrodes nerve to cause muscle twitch (Fig.3). 7. Apply a higher voltage of about 6 V to stimulate the nerve again. Fig.3 Applying an electric current 84.........Practical work Note 1. The frog should not be killed by over anaesthesia with chemicals, as this will damage the irritability of nervous system of the frog. 2. The frog can be killed by the following methods: Method 1 • use a needle pin to destroy the nervous tissues of the brain; • insert the needle pin into the spinal cord to damage its nervous tissues, until both legs become straightened. Method 2 • cut the upper jaw of the frog with a bone cutter to expose the spinal cord; • insert a needle pin in the spinal cord to damage the nervous tissues until both legs become straightened. 3. A 2 V electrical stimulus is strong enough to cause depolarisation of the nerve fibre and evoke an action potential as well as a single twitch of muscle contraction.