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Electrical stimulation (10 Hz) led to a threefold increase in [<jats:sup>3<\/jats:sup>H]citrulline formation. Removal of calcium from the medium or addition of N<jats:sup>G<\/jats:sup>-nitro-l-arginine strongly inhibited both spontaneous and electrically induced production of [<jats:sup>3<\/jats:sup>H]citrulline. TTX reduced the electrically induced but not spontaneous [<jats:sup>3<\/jats:sup>H]citrulline formation. The electrically induced formation of [<jats:sup>3<\/jats:sup>H]citrulline was diminished by (+)-tubocurarine and mecamylamine and enhanced by scopolamine, which suggests that endogenous ACh inhibits, via muscarinic receptors, and stimulates, via nicotinic receptors, the NO synthesis in the myenteric plexus. The GABA<jats:sub>A<\/jats:sub>receptor agonist muscimol and GABA also reduced the electrically evoked formation of [<jats:sup>3<\/jats:sup>H]citrulline, whereas baclofen was without effect. Bicuculline antagonized the inhibitory effect of GABA. 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