The morphology and distribution of NADPH diaphorase reactive neurons was studied in the normal human hypothalamus. Reactive neurons were divided into three categories on the basis of perikaryal size. Small neurons (8-20-mu-m) were oval or fusiform, and pale staining. Intermediate neurons (20-30-mu-m) were fusiform, triangular or pyramidal with a wide range of staining intensity. Large neurons (> 30-mu-m) were triangular or pyramidal with moderate to dark staining. Reactive neurons were found in four major regions: medial preoptic, ventromedial, lateral, and perifornical. Scattered positive neurons were found in several other hypothalamic areas. Reactive fibers were present in the supraoptic decussation, medial forebrain bundle, and stria medullaris thalami. The localization of NADPH diaphorase neurons in hypothalamic nuclei affected by Alzheimer's disease and other degenerative disorders suggests that further studies of this neuronal subset are warranted.