3,4-Methylenedioxymethamphetamine (MDMA) use has been associated with a decline in various aspects of mnemonic function in humans. We therefore postulated that MDMA-induced damage of serotonergic nerve terminals would alter hippocampal processing. Seven days following treatment with MDMA (2 x 20 mg/kg sc, given 12 h apart), rat spatial learning and memory were tested utilizing the Morris water maze (MWM). No statistical differences were found in MWM platform acquisition latency or pathlength between controls and MDMA-treated animals. Probe trials revealed significantly higher proximity score averages and significantly reduced preference for the target quadrant in the MDMA-treated animals. MDMA treatment resulted in significant reduction (34%) in hippocampal serotonin (5-HT) levels 14 days after initial treatment. The findings of this study demonstrate that hippocampal serotonergic lesions induced by MDMA may be ostensibly linked to a reference memory deficit in rats tested with the MWM.