The distribution of gonadotropin-releasing hormone (GnRH) was studied in the brain of adult female rats with three immunocytochemical techniques using antisera to unconjugated synthetic GnRH and to GnRH conjugated with limpet hemocyanin. GnRH was found in nervous tissue surrounding blood vessels of the organum vasculosum of the lamina terminalis. In the median eminence it occurred in nervous tissue associated primarily with the tuberoinfundibular sulci throughout their extent. Cephalic to the pars tuberalis GnRH often spread across the median eminence from sulcus to sulcus. Caudally, with widening of the median eminence, GnRH occurred dorsal to the tuberoinfundibular sulci, and especially in the external lamina medial to the sulci. A broad median zone of the median eminence was rather free of GnRH. GnRH was most concentrated in the region of continuity between the dorsolateral walls of the infundibulum and floor of the third ventricle where the tuberoinfundibular sulci are deep. Caudal to the infundibulum GnRH was disposed in a flat zone through the cephalic portion of the floor of the mammillary recess. In the median eminence GnRH appeared to be located in axons that terminated there. The amount of demonstrable GnRH varied significantly from rat to rat. The distributions of GnRH as revealed by use of antisera to unconjugated and conjugated GnRH were essentially the same. The apparent order of sensitivity of the immunocytochemical methods was: the peroxidase-antiperoxidase (PAP) (Sternberger et al.) procedure greater than the immunoglobulin-enzyme bridge (Mason et al.) procedure smaller than the conjugated antibody (Nakane and Pierce) procedure.