To investigate the possibility that serotonin plays a role in the sexually dimorphic development of a nucleus in the medial preoptic area of the rat brain, p-chlorophenylalanine, an inhibitor of serotonin biosynthesis, was administered to pregnant dams from day 8 of gestation until parturition. This treatment did not alter plasma steroid levels but increased the volume of the sexually dimorphic nucleus in female neonates to that of control males. Thus, serotonin is implicated as a neurochemical which may be involved in the sexually dimorphic development of the preoptic area.