In female rats the luteinizing hormone (LH) is timed by the circadian system and is followed by a display of intense, estrogen-induced running behavior. This proestrous running on the night of ovulation can be used as a marker of the estrous cycle. Entrainment of the mammalian circadian system by exogenous melatonin (MT) has been demonstrated only in the activity rhythms of male rats. The present experiments were designed to study the effect of daily MT injections on activity rhythms and proestrous running of female rats in 1) continuous dim white light (LL) and 2) continuous darkness (DD). In LL, MT injections (50 micrograms/kg or 1 mg/kg) had no discernible effect on activity rhythms. In DD, four of the six MT-treated rats (100 micrograms/kg) entrained to the injection, and a fifth animal showed phase advances in its activity rhythm when onset of activity passed through injection time. The sixth animal was not injected with MT at activity onset time. None of the six control animals showed either effect. MT had no effect on the length of the estrous cycle. Thus MT injections can entrain circadian rhythms of activity and proestrous running in female rats in DD but not in LL.