Direct evidence for a release of acetylcholine from the myenteric plexus of guinea pig small intestine by substance P.

Myenteric plexus-longitudinal muscle strips from guinea pig small intestine were labeled with [3H]choline while under continuous field stimulation. Release of newly synthesized [3H]acetylcholine was studied in the presence of substance P afterwards. Substance P evoked release of [3H]acetylcholine in a dose-related fashion. Both tetrodotoxin and [D-Pro2,D-Phe7,D-Trp9]substance P, a synthetic antagonist of substance P, completely blocked this release and provided evidence for a cholinergically mediated mechanism of substance P on enteric neurons.