The objective was to determine the distribution of growth hormone-release-inhibiting hormone (somatostatin) in the rat brain using the peroxidase-antiperoxidase immunocytochemical method with antisera prepared against unconjugated, synthetic somatostatin. Somatostatin occurred in low quantity in the organum vasculosum of the lamina terminalis. It was present throughout the full length of the median eminence and occupied the entire width between the tuberoinfundibular sulci. Most somatostatin was located in the dorsal portion of the external lamina, and the amount varied according to the mediolateral position. The bodies labeled for somatostatin were most often granules; occasionally they appeared as clusters of granules that seemed to be membrane-enclosed. Some of these bodies appeared to be portions of axons. Many of the larger bodies were arranged alongside tanycytes, but no label was distributed generally in tanycyte cytoplasm. Somatostatin was highly concentrated in the proximal one-quarter of the infundibular stem and appeared in lower concentration throughout the distal portion of the stem. It was absent from the pars nervosa and pars intermedia of the pituitary gland. The distribution of somatostatin in the median eminence differed considerably from that of gonadotropin-releasing hormone. Somatostatin was identified in the ventromedial and/or dorsomedial hypothalamic nuclei of only two animals. Here it was probably located in axons that terminated on neuronal cell bodies but also may have been present in a restricted portion of the perikaryonal cytoplasm.