Numerous biologically active peptides are produced by specialized cells of the gastrointestinal tract. Most of these peptides are also produced outside the gut, and current evidence suggests that they not only regulate digestive events per se but also participate in many other regulatory mechanisms working as hormonal, paracrine, and neuronal messengers. The physiological functions of the gastrointestinal peptides are yet very incompletely known. Immunocytochemical tracing of the destinations of neuronal and paracrine cell processes may, together with available physiological and biochemical data, provide valuable clues to the sites of actions of many of the novel regulatory peptides. Moreover, immunocytochemistry has given evidence for the occurrence of multiple secretory peptides in certain endocrine cell types and suggested that certain peptides simultaneously may be secreted by multiple endocrine, paracrine, and neural cell types. This review emphasizes the continued need for concerted cytochemical, physiological, and biochemical studies of the sites of synthesis, secretion, and action of gastrointestinal regulatory peptides.