Cholecystokinin is a classical gastrointestinal hormone that is produced by discrete endocrine cells of the upper small intestine. Cholecystokinin is produced in various molecular forms that result from differences in posttranslation processing of a single gene product. Cholecystokinin is secreted from the intestine in response to the ingestion of food. We observed that specific dietary substances increase the rate of transcription of the cholecystokinin gene and stimulate cholecystokinin release in rats. In contrast the paracrine transmitter, somatostatin, inhibits dietary-stimulated cholecystokinin secretion and lowers intestinal mRNA levels. Evidence that cholecystokinin gene expression is not necessarily linked to hormone secretion is supported by the observation that the neuropeptide, bombesin, stimulates cholecystokinin release but does not modify intestinal cholecystokinin mRNA levels. To examine the intracellular messengers that might regulate the cholecystokinin cell directly, we developed an in vitro method for studying cholecystokinin release from isolated intestinal mucosal cells. In this perifusion system, cholecystokinin release was stimulated by membrane depolarizing concentrations of KCl (50 mmol/L), the calcium ionophore A23187 (1 mumol/L), and the cAMP analogue dibutyryl cAMP (1 mumol/L). Biologically active cholecystokinin was also released in a dose-dependent manner by the peptide transmitters, bombesin and monitor peptide. These findings indicate that neurotransmitters and hormones may directly regulate the cholecystokinin cell and suggest that the phosphoinositide and adenylate cyclase cascades mediate stimulated-cholecystokinin secretion.