The present study examines the effects of physiological levels of progesterone (P) on copulatory behavior in sexually naive male rats. Two weeks after gonadectomy males were implanted with either empty Silastic capsules (BL) or Silastic capsules containing testosterone (T), P, or both (P+T). When tested with an estrous female, all of the gonadally intact males (intact) and none of the BL controls exhibited mounting/intromission behaviors. Mounting was observed in 75% of the T-alone males. More than half (64%) of the P-alone males and 100% P+T males exhibited mounting. In most cases, mounting was followed by intromission responses. Subsequently, intact and gonadectomized males received daily injections of the P antagonist RU486 along with hormone treatment. After receiving RU486, only 63% of the intact males and 71% of the T-alone males mounted successfully. The facilitatory effects of P on copulatory behavior were completely abolished by RU486 treatment. The present studies provide the first evidence in mammals suggesting that P-dependent mechanisms influence neurochemical pathways involved in copulation.