Vasotocin (VT) and vasopressin control many endocrine and neuroendocrine functions, including the regulation of reproductive behaviors. In the roughskin newt (Taricha granulosa), VT administration can enhance courtship behaviors in males and egg-laying behaviors in females. This study used immunohistochemistry to investigate whether there are sex differences in VT in specific brain areas, and whether these differences persist in nonbreeding animals. Numbers of VT immunoreactive (ir) cell bodies were counted in males and females collected in February, April, June, and August. Radioimmunoassay of plasma samples confirmed that testosterone and 5α-dihydrotestosterone concentrations were higher in males than females, and that 17β-estradiol concentrations were higher in females than males. In 11 brain areas, no sexual or seasonal differences in the number of VTir cells were found. But in 3 brain regions—the bed nucleus of the stria terminalis (BNST), the nucleus amygdalae dorsolateralis (AMYG), and the anterior preoptic area (aPOA)—there were significantly greater numbers of VTir cells in males than in females, and these differences did not change seasonally. In the aPOA, an area important to male sex behaviors, the sexual dimorphism in VTir was particularly pronounced. In four brain regions, there were significantly greater numbers of VTir cells in females than males, but only in specific seasons. In April-collected (breeding) animals, more VTir cells were found in females than in males in the populations of VT cells within the pars dorsalis hypothalami and ventromedial hypothalamus, brain regions frequently associated with stress responses and female mating behaviors. In August-collected (nonbreeding) animals, more VTir cells were found in females than in males, in the region of the bed nucleus of the decussation of the fasciculus lateralis telencephali and in the nucleus visceralis superior, nucleus isthmi region. Significantly greater numbers of VTir cells were observed in the magnocellular preoptic area of males and females collected in February. These results indicate that the functional interactions between gonadal steroid hormones and VT are complex and appear to involve site-, sex-, and season-specific regulatory mechanisms. Furthermore, it seems likely that populations of VT neurons in the BNST, AMYG, and aPOA are involved in regulating male-specific behaviors, and that the VT neurons in the pars dorsalis hypothalami/ventromedial hypothalamus may be involved in female-specific behaviors.