The regional and topographic distribution of substance P-containing nerve fibers in the human and feline intestinal wall was studied by immunocytochemistry and radioimmunoassay. The concentration of substance P was measured in the different layers of the duodenum, jejunum, ileum, and colon. In both humans and cat, substance P fibers were fairly numerous, and the substance P concentration was comparatively high in the smooth muscle layer, including the myenteric ganglia. In humans, but not in cat, substance P fibers were numerous, and the substance P concentration was also high in the mucosa. Substance P-containing nerve cell bodies were observed in the myenteric ganglia of both species. In the submucous ganglia, such nerve cell bodies were seen in the human intestine only, suggesting that they represent the origin of the numerous mucosal substance P fibers in this species. Previous studies have revealed a relative paucity of substance P fibers in the intestinal mucosa of several mammals, such as mouse, rat, and pig. The cat can now be added to those having few mucosal substance P fibers, whereas humans seem to be notably rich in such fibers, suggesting that substance P may play a role in the regulation of mucosal functions in the human intestine.