Effects of 5-hydroxytryptamine (5-HT) and related agonists and antagonists on formation of cAMP were determined for enzymatically dissociated ganglia from the myenteric plexus of the guinea-pig small intestine. Formation of cAMP was stimulated by 5-HT in both dose- and time- dependent manners. The stimulatory action of 5-HT was suppressed by the 5-HT1P antagonist, renzapride, but not by the 5-HT3 antagonist, tropisetron (formerly ICS 205-930). Neither renzapride nor cisapride increased the levels of cAMP. Levels of cAMP were suppressed by concentrations of tropisetron greater than 1.0 microM. Levels of cAMP were unaffected by the 5-HT3 agonist, 2-methyl-5-hydroxytryptamine. A putative 5-HT4 receptor agonist, 5-methoxytryptamine, stimulated formation of cAMP, but to a lesser extent than 5-HT. We conclude that 5-HT acts to stimulate adenylate cyclase and the formation of cAMP in myenteric ganglia. The 5-HT1P receptor is the most likely subtype involved in 5-HT action on cAMP formation.