Intraperitoneal administration of the non-competitive NMDA receptor antagonists (5R,10S)-(+)-5-methyl-10,11-dihydro-5H-dibenzo[a,d] cyclohepten-5,10-imine (MK-801, 0.25 and 0.5 mg/kg) and 1-(1-phenylcyclohexyl)piperidine (PCP, 5 and 10 mg/kg) increased the extracellular concentration of acetylcholine in rat hippocampus but not striatum. In contrast, R-(+)-3-amino-1-hydroxypyrrolid-2-one (R(+)-HA-966, 30 and 60 mg/kg), an antagonist at the glycine modulatory site of the NMDA receptor, did not affect acetylcholine efflux in either region. (+/-)-3-[2-Carboxypiperazin-4-yl]-propyl-1-phosphonic acid ((+/-)CPP, 10 mg/kg) and cis-4-(phosphonomethyl)piperidine-2-carboxylic acid (CGS19755, 5 mg/kg), competitive antagonists at the glutamate agonist site of the NMDA receptor, marginally increased hippocampal acetylcholine efflux. Pretreatment with R(+)-HA-966 (60 mg/kg) or (+/-)CPP (10 mg/kg) attenuated the increase of hippocampal acetylcholine efflux by MK-801 (0.5 mg/kg). However, prior administration of CGS19755 (5 mg/kg) prolonged the MK-801-induced increase of hippocampal acetylcholine efflux. Results demonstrate differential effects on hippocampal and striatal acetylcholine efflux of antagonists at different sites on the NMDA receptor complex and are discussed in relation to previously described effects of these drugs on mesolimbic dopamine function.