In golden hamsters, vasopressin (AVP) microinjected within the ventrolateral hypothalamus (VLH) facilitates offensive aggression. As serotonin is known to inhibit offensive aggression, we decided to test whether AVP-facilitated behavior is also inhibited by serotonin treatment. Testosterone-treated male golden hamsters received IP injections of fluoxetine, a serotonin reuptake inhibitor, or vehicle 1 h prior to AVP microinjections within the VLH. The animals were tested for offensive aggression in a resident-intruder model after the microinjections, and the results were compared between groups. Pretreatment with fluoxetine inhibited AVP-facilitated offensive aggression. Only one out of nine fluoxetine-treated animals attacked and bit the intruders, compared to six out of seven vehicle-treated animals. Furthermore, we also confirmed by in vitro autoradiography that the VLH contains vasopressin V(1) and serotonin 5-HT1B receptors. Therefore, it is possible that serotonin may inhibit AVP-facilitated offensive aggression by acting directly at the level of the VLH as well as at other sites.