Localization of oxytocin and vasopressin binding sites has so far been studied in the rat brain by means of film autoradiographs. The availability of selective iodinated ligands with high specific activity allowed us to develop the histoautoradiographical technique and to reinvestigate at the microscopic scale the distribution of these sites in the hypothalamus. Most oxytocin binding sites were localized in delimited nuclei, e.g., the medial preoptic, the ventromedial, the ventral premammillary, the supramammillary, and the medial mammillary nuclei. In addition, a weak diffuse specific labeling occurred in the medial preoptic and the anterior hypothalamic areas. The vasopressin binding sites (of the V1a type) were detected in delimited nuclei, e.g., the suprachiasmatic, the stigmoid, and the arcuate nuclei, but they were also diffusely distributed in the lateral hypothalamic and the dorsochiasmatic areas. The locations of neurohypophysial peptides binding sites detected by light microscopy are compared with those previously obtained by film autoradiography.