Development of Tolerance to Ethanol in Cultured Adrenal Chromaffin Cells

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Abstract
Dissociated bovine adrenal chromaffin cells in culture were utilized to study the mechanisms for development of cellular tolerance to ethanol. Three days after plating, cells were grown in either control medium or medium containing 200 mM ethanol for periods up to 6 days. Catecholamine release induced by carbachol, 80 mM K+, BAY K 8644, and veratrine was studied in ethanol-treated and untreated cultures. The presence of ethanol in vitro was inhibitory to all these stimuli in untreated cell cultures, carbachol-induced release being most sensitive (IC50 approximately 30 mM). After growth in ethanol for 6 days, carbachol-induced catecholamine release was completely resistant to the inhibitory effects of ethanol. The resistance to ethanol of carbachol-induced catecholamine release from cells grown in medium containing ethanol also extended to the inhibitory effects of butanol. The results show some similarities with changes observed in the central nervous system during the development of ethanol tolerance, and it is suggested that this model system may be useful in examining the mechanisms by which tolerance occurs.