Estrogen is known to modulate the behavioral response to cocaine; however the mechanisms by which this is accomplished is unknown. In this study we examine one possible candidate, the endogenous opioid system. Adult Sprague-Dawley rats were ovariectomized (OVX), half received Silastic implants with estradiol benzoate (OVX-EB), the other half received empty implants (OVX). After 1 week, spontaneous locomotor and stereotyped activity was measured for 60 min using an automated system. On day 2, locomotor activity was recorded for 30 min. Rats were injected with saline (SAL) or naloxone (NAL) (2 mg/kg, i.p.) and activity measured for the next 20 min. Each of these groups were further subdivided, one that received a saline injection (SAL) and another that received a cocaine injection (COC) (15 mg/kg, i.p.). Locomotor and stereotyped activities were recorded for 60 min. This resulted in the following injection groups: SAL-SAL, NAL-SAL, SAL-COC and NAL-COC. During habituation, OVX rats displayed an overall higher level of activity than OVX-EB rats. Similar to what is observed in males, naloxone significantly reduced locomotion and stereotyped behavior but only in OVX rats. Estrogen administration to OVX rats abolished the effect of naloxone. Surprisingly, when naloxone was administered prior to cocaine, an increase in cocaine-induced locomotor and stereotyped activity was observed, but only in OVX-EB rats. These results indicate that opioid modulation of cocaine-induced locomotor and stereotype activity in the female differs from that reported in the male. In addition in the female, the effect of opioids on cocaine-induced locomotor behavior is dependent on plasma levels of estrogen.