Systemic injections of morphine impair performance in memory tests. Glucose administration ameliorates memory deficits produced by morphine treatment. The memory impairments induced by morphine may be related to opioid inhibition of acetylcholine release with reversal of this effect by glucose. The present experiment determined whether: (1) systemic morphine treatment decreases acetylcholine output in the hippocampal formation; and (2) systemic glucose administration attenuates the effect of morphine treatment. Employing microdialysis, samples were collected at 12-min intervals and assayed for acetylcholine using HPLC with electrochemical detection. Morphine (10 mg/kg)/saline injections resulted in an immediate decrease in acetylcholine output (20-35%) that was observed up to the third postinjection sample (36 min). Glucose (100 mg/kg) administered concurrently with morphine attenuated the reduction in acetylcholine output in the second and third samples. These findings suggest that glucose may attenuate morphine-induced memory impairments by reversing a decrease in acetylcholine output produced by morphine.