Inhibition of Muscarinic Receptor-Stimulated Glial Cell Proliferation by Ethanol

Abstract: Acetylcholine and other muscarinic agonists stimulate the proliferation of rat cortical astrocytes and 132 1N1 human astrocytoma cells by activating muscarinic m₃ cholinergic receptors. Ethanol was a potent inhibitor of carbachol-stimulated proliferation, measured by ³H]thymidine incorporation, with an IC₅₀ of 10 m M . On the other hand, basal and serum-stimulated proliferation of astrocytes and astrocytoma cells was inhibited by ethanol with lower potency (IC₅₀ = 200–250 m M ). Concentration-response experiments with carbachol, in the presence of 10 m M ethanol, suggested that inhibition of proliferation by the alcohol was of the noncompetitive type. Experiments with acetaldehyde and with the alcohol dehydrogenase inhibitor 4-methylpyrazole suggested that the inhibitory effect of alcohol was due to ethanol itself and not to its metabolite acetaldehyde. Proliferation of astrocytoma cells induced by carbachol and the inhibitory effects of ethanol were also confirmed by flow cytometry using the 5-bromodeoxyuridine-Hoechst 33258 method. Ethanol (10 m M ) had no effect on proliferation induced by 50 µg/ml insulin and 100 ng/ml platelet-derived growth factor BB; on the other hand, the mitogenic effect of 1 m M histamine, 100 U/ml interleukin-1, and 100 ng/ml 12- O -tetradecanoylphorbol 13-acetate were inhibited by ∼50%. These results indicate that proliferation of glial cells induced by muscarinic agonists is especially sensitive to the inhibitory effect of ethanol. This action of ethanol may be relevant to its developmental neurotoxicity, particularly microencephaly, which is one of the common features of the fetal alcohol syndrome.