Golden hamsters, as compared to rats, lack several parvicellular vasopressinergic cell groups, particularly sexually dimorphic populations. We decided to test the possibility that magnocellular vasopressinergic neurons are subjected to sexual differences in hamsters, as they are known to display vasopressin (AVP)-dependent sexually dimorphic behaviors. The distribution of magnocellular vasopressinergic neurons was mapped and compared between males and females. Approximately 50% more vasopressin-immunoreactive (AVP-ir) neurons were counted in males within the medial and lateral divisions of the supraoptic nucleus. Furthermore, levels of AVP extracted from the hypothalamus and the pituitary gland were three to four times higher in males than in females. Finally, hypothalamic extracts from a male and a female hypothalamus were fractionated by HPLC and assayed for AVP immunoreactivity. Immunoreactivity from each extract had the same retention time as synthetic AVP standards; and the levels were twice as high in the male. These results support the existence of sexual differences in the magnocellular vasopressinergic system in golden hamsters. These differences appear to be related to previously reported sexual differences in AVP secretion from the neurohypophysis.