Choline acetyltransferase activity was measured in rats treated with daily injections of ethanol (0.1 g/kg body wt) and or dexamethasone (1 mg/kg body wt) for 5 consecutive days. Ethanol produced a biphasic reduction of choline acetyltransferase activity in rat cerebral cortex, which at most time points was further decreased by simultaneous injection of dexamethasone. Kinetic studies of cortex choline acetyltransferase activity in rats that had received 5 daily injections of ethanol or ethanol and dexamethasone indicated that the observed reduction in enzyme activity was due to an apparent reduction in affinity (K(m)) of the enzyme for acetyl coenzyme A with no significant change in the total amount of enzyme present (Vmax). This finding has implications with respect to the use of choline acetyltransferase as a marker for cholinergic neurons, and for the understanding of the regulation of choline acetyltransferase activity in the brain.