Release of an insulin‐like peptide from perfused extirpated cat legs in response to electrical stimulation of the sciatic and brachial nerves and to administration of ACh, bombesin, oxytocin and glibenclamide

Abstract

The vascular system of extirpated cat legs was perfused with Tyrode's solution and insulin-like immunoreactivity (ILI) levels were determined in the perfusate with radioimmunoassay. During unstimulated conditions perfusate levels of ILI were almost undetectable. However, in response to electrical stimulation of the sciatic or brachial nerves (within a wide range of stimuli 5-40 V, 2-20 Hz and 0.2-40 ms) 1-20 ng of ILI was recorded in the perfusate. Blockers of cholinergic and adrenergic transmissions added to the perfusate did not influence the output of the ILI induced by nerve stimulation. Furthermore, after administration of acetylcholine (ACh) (0.1 and 10 micrograms kg-1), oxytocin (0.5 and 5 IU kg-1), glibenclamide (25 and 100 micrograms kg-1) and bombesin (100 and 500 micrograms kg-1) to the cat leg preparation, ILI appeared in the perfusate in amounts similar to those induced by electrical stimulation of the nerves. When subjected to high-performance liquid chromatography (HPLC) the insulin-like peptide detected in the cat leg perfusate following nervous stimulation, or administration of oxytocin and glibenclamide, co-eluted with a bovine insulin standard. We have previously shown that some peripheral nerves of the cat, such as the sciatic, brachial and vagal nerves, contain an insulin-like peptide with HPLC characteristics similar to the bovine insulin standard. It is therefore possible that the insulin-like peptide released from the isolated cat leg preparation by the above-mentioned stimuli derives from this nervous pool of insulin. Alternatively, the insulin-like peptide emanates from the striatal muscles innervated by the sciatic and brachial nerves, since also muscles have been shown to contain an insulin-like peptide.