Release from and accumulation in tissue slices of some neurotransmitters under acute ethanol in naive rats and in long‐term voluntarily ethanol drinking rats were investigated. Slices of the rat caudatoputamen were prelabeled with [3H]choline and release of [3H]acetylcholine was stimulated through either N‐methyl‐d‐aspartate (NMDA) receptors or strychnine‐sensitive glycine receptors. Ethanol in vitro at 2%, 4%, and 6% (34 mM, 68 mM, and 102 mM, respectively) concentration‐dependently depressed the maximum effect of the concentration‐response curve of NMDA in naive rats. In contrast, voluntary ethanol consumption over months led to a significantly enhanced NMDA receptor response characterized by an increase in the maximum effect of the concentration‐response curve. The glycine receptor‐mediated release of [3H]acetylcholine, which is inhibited by acute ethanol in a competitive‐like fashion, was not changed in animals that ingested ethanol over months. Electrically evoked release of [3H]noradrenaline ([3H]NA) and its presynaptic modulation by morphine through μ‐opioid receptors in neocortical slices of the rat, preloaded with [3H]NA, was nearly identical in both ethanol‐naive rats and in ethanol drinking rats. The accumulation of [3H]γ‐aminobutyric acid in rat cerebellum tissue was neither affected by acute ethanol in vitro nor after chronic ethanol consumption. In summary, long‐term voluntary ethanol intake caused a significant increase in NMDA receptor function in the rat caudatoputamen, but did not result in changes in glycine‐evoked [3H]acetylcholine release of electrically evoked [3H]NA release modulated by morphine or cerebellar [3H]γ‐aminobutyric acid accumulation.