Male rats that had cohabited with ovariectomized females for 2 weeks became more aggressive toward male intruders after a novel estrous female had been placed in their home cages for a period of 4 hr on the previous day. No increase in aggression was seen in males exposed to anestrous females. Genital anesthesia did not attenuate the female-enhanced aggression effect. By contrast, no enhancement of aggression was observed in long-term castrated males after 24 hr of exposure to estrous females. Rats with bilateral electrolytic lesions in the medial preoptic area showed a normal female-enhanced aggression effect. The observations that exposure to females facilitates aggression in males subjected to genital anesthesia and in males with preoptic lesions raise the possibility that copulatory cues are not always required for the effect.