The potent vasoconstrictor endothelin was originally isolated from vascular endothelial cells but has since been found in several other tissues. The aim of this study was to establish whether endothelinlike immunoreactivity occurs in human enteric nerves and to identify endothelin binding sites using immunocytochemical and in vitro autoradiographic techniques. Endothelinlike immunoreactivity was localized to nerve bundles throughout the colon and to most of the ganglion cells of the two major plexuses, many of which costored vasoactive intestinal polypeptide. High-affinity (dissociation constant = 0.35 +/- 0.014 nmol/L; mean +/- SEM) binding sites for endothelin 1, with an apparent binding capacity of 92 +/- 6.3 amol/mm2 (mean +/- SEM), were demonstrated in the myenteric plexus, with less dense binding being found in the submucous plexus, mucosa, muscle layers, and blood vessel walls. Competition data suggested two populations of binding sites, both showing high affinities for endothelins 1 and 2, vasoactive intestinal constrictor, and sarafatoxin b but differentiated by their affinity for endothelin 3 and sarafatoxin c. This study provides evidence that endothelin is a neuropeptide in the human intestine with binding sites on neural plexuses and mucosa, suggesting a role in the modulation of intestinal motility and secretion.