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The time course for the re-acquisition of membrane tolerance to the disordering effects of ethanol in vitro has been determined for liver microsomes obtained from chronically ethanol-fed rats that were withdrawn from ethanol for 2-4 days (during which tolerance is lost) followed by resumption of ethanol feeding. Naive rats require 28-35 days of chronic ethanol feeding to develop membrane tolerance. Microsomal membranes regain partial sensitivity to ethanol disordering after 2-3 days of withdrawal and regain the complete sensitivity observed in membranes from untreated control rats after 4 days of withdrawal. The period of ethanol re-feeding required for the re-acquisition of membrane tolerance was dependent on the withdrawal period, with tolerance appearing sooner if the withdrawal period was shorter. The time course for the re-development of tolerance in previously tolerant animals was considerably faster (4-14 days) than in naive rats being administered the ethanol diet for the first time (35 days). Microsomes from rats that were withdrawn for 2 days (which retained partial tolerance) and then re-fed the ethanolic-diet required only 4 days to re-acquire membrane tolerance. Microsomes from rats withdrawn 3 days required 8 days and those withdrawn 4 days required 15 days for full tolerance to re-develop. The same time-course for the re-acquisition of membrane tolerance was observed in either intact microsomes or in liposomes prepared from extracted microsomal total phospholipids. Phosphatidylinositol (PI) has previously been reported to be responsible for conferring membrane tolerance to liver microsomes in ethanol-fed rats (Taraschi, T.F., Ellingson, J.S., Wu, A., Zimmerman, R. and Rubin, E. (1986) Proc. Natl. Acad. Sci. USA 83, 9398-9402). The time course for re-acquisition of membrane tolerance by liver microsomes following ethanol withdrawal and resumption of ethanol feeding correlated with the ability of PI to confer tolerance.
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