The analgesic effect of buprenorphine is mediated via the mu opioid receptor (MOP). In the present study, using mice lacking the MOP and their wild-type littermates, we determined the role of the MOP in buprenorphine-induced locomotor stimulation and conditioned place preference (CPP). Buprenorphine (3 mg/kg) increased motor activity in wild-type but not in MOP knockout mice, showing the motor stimulatory action of buprenorphine is mediated via the MOP. When the mice were given the same treatment once daily for 5 consecutive days and challenged with buprenorphine on day 11, the motor stimulatory action of buprenorphine was enhanced in wild-type but not in MOP knockout mice, showing sensitization developed to the motor stimulatory action of buprenorphine and this phenomenon was mediated via the MOP. Likewise, buprenorphine induced CPP in wild-type mice after four alternate-day saline/buprenorphine (3 mg/kg) injections paired with olfactory and visual cues. However, buprenorphine failed to induce CPP in MOP knockout mice. In contrast, amphetamine (1 mg/kg) induced a comparable CPP in wild-type and MOP knockout mice. Together, the present results suggest that the ability of buprenorphine to increase motor activity and induce locomotor sensitization and CPP is mediated via the MOP.