Numerous experiments suggest that perinatal exposure of male vertebrates to testosterone (T), or its estrogenic metabolites, masculinizes aspects of coital function, including males' characteristic preference to seek out and mate with a female as opposed to another male conspecific. Other research has shown that this perinatal action of sex steroids also masculinizes aspects of neuronal morphology in the medial preoptic area/anterior hypothalamus (mPOA/AH). We asked whether neurons of the mPOA/AH contribute to males' preference to mate with a female. The ferret is an ideal species in which to ask this question. When tested in a T-maze after gonadectomy and treatment with estradiol benzoate (EB), female ferrets prefer to approach and receive neck grips from a stimulus male whereas males prefer to approach and neck grip an estrous female. In the minority of trials when EB-treated males approach a stimulus male, they occasionally receive a neck grip to which they display receptive postures as opposed to agonistic behaviors. In Experiment 1 castrated, EB-treated male ferrets which received bilateral infusions of the NMDA excitotoxin, quinolinic acid aimed at the dorsomedial POA/AH, preferred to approach a stimulus male significantly more often than groups of control males which either received a sham lesion, received a unilateral mPOA/AH lesion or in which bilateral infusions of quinolinic aci produced no histologically detectible excitotoxic damage to the mPOA/AH. Males with bilateral mPOA/AH lesions also displayed neck gripping on a significantly lower percentage of trials than control males when they approached the stimulus female. Ovariectomized, EB-treated female ferrets with bilateral mPOA/AH lesions, like control females, preferred to approach and receive neck grips from a stimulus male. The males used in Experiment 1 had never experienced circulating levels of T characteristic of the breeding season. Therefore, in Experiment 2 prepubertally gonadectomized males and females were treated chronically with a high dose of T propionate (TP) and tested several times with a receptive female prior to brain surgery. Males which received bilateral excitotoxic lesions of the mPOA/AH neck gripped and mounted stimulus females significantly less than control males. Again, when given EB followed by T-maze tests of partner preference, males with bilateral mPOA/AH lesions, like sham-operated female controls, preferred to approach the sexually active stimulus male significantly more often than control males, which preferred to approach the stimulus female. Our results suggest that neurons in the mPOA/AH play an important role in the integration of sensory cues which determine heterosexual partner preference in the male ferret, in addition to facilitating masculine coital performance.