Effect of ethanol tolerance on release of acetylcholine and norepinephrine by rat cerebral cortex slices

The release of acetylcholine (ACh) by rat cerebral cortex slices, with and without electrical stimulation, and the effect of ethanol (EtOH) on this release were examined during the acquisition and loss of EtOH tolerance in vivo. ACh was measured by pyrolytic monodemethylation and gas–liquid chromatography. Electrical stimulation of control slices in medium containing diisopropyl phosphofluoridate (1.26 μM) and atropine (0.3 μM) increased ACh release by 88 ± 12%. Addition of 0.11 M EtOH to the medium had negligible effect on ACh release from unstimulated slices, but reduced the effect of stimulation to 51 ± 10%. After chronic treatment with EtOH by gavage or in a liquid diet, rats became tolerant to EtOH in vivo as shown by reduced impairment on the moving belt test. Slices from tolerant rats showed increased release of ACh in response to electrical stimulation and less inhibition of this response by added EtOH. The changes had disappeared by 2 weeks after cessation of EtOH treatment.Similar findings were obtained by measurement of release of [14C]ACh from slices preloaded with [14C]choline, except that electrical stimulation in the absence of EtOH appeared to cause a smaller increase in slices from chronic EtOH animals than from controls. This may reflect differences in isotope dilution. Release of [3H]norepinephrine was less affected by EtOH than that of ACh. The findings suggest that tolerance to EtOH is accompanied by increased ACh release by cortical neurones, as well as decreased direct inhibitory effect of EtOH on this, but do not permit any conclusion about the relative importance of such changes in various parts of the brain.