The distribution of endothelin-like immunoreactivity was examined in normal rat kidneys using the immunoperoxidase technique. A specific polyclonal antibody to endothelin 1, which recognized endothelin 1 and its precursor molecule, big endothelin, was raised in rabbits. Immunoperoxidase staining revealed specific endothelin-like immunoreactivity in the renal cortex, medulla, and papilla. Immunostaining density was greatest in the renal papilla where staining was predominantly localized to the vasa rectae of the distal nephron segments. Cytoplasmic immunostaining was noted focally in collecting duct cells in the renal papilla. In the renal medulla, intense immunostaining was identified in the vasa rectae. Cortical immunostaining was localized to the endothelial surfaces of arcuate arteries, veins, arterioles and peritubular capillaries. Glomerular immunostaining followed a capillary loop distribution and appeared to be predominantly localized to endothelial cells with smaller amounts of reaction product overlying the mesangium. The most proximal portion of the proximal tubule brush border and papillary collecting duct epithelium demonstrated focal endothelin-like immunostaining. We conclude that endothelin-like immunoreactivity is widely distributed in renal tissue compatible with important autacrine and paracrine actions in the kidneys.