Ethanol causes decreased partitioning into biological membranes without changes in lipid order

Abstract

One of the adaptive responses of cell membranes to chronic ethanol consumption is the acquisition of a resistance to fluidization or disordering of the lipids by ethanol in vitro and a reduced partitioning of ethanol into the membrane (membrane tolerance). The degree to which the effects on partitioning and lipid disordering share common features has not previously been explored and in addition the relevance of the value of lipid order in the absence of added ethanol (baseline lipid order) to membrane tolerance has not been established. The location in the bilayer and the nature of the modification underlying these effects is also unknown. The effect of chronic ethanol treatment was examined using 5-doxyl decane as a model hydrophobic compound. Its partitioning into the membranes was determined by utilizing its ability to quench fluorophores (1,6-diphenyl-2,3,5-hexatriene and 3- and 12-anthroyl stearates) by collisional quenching. The partition coefficient of 5-doxyl decane into the bilayer central region was reduced as a result of the chronic ethanol treatment. The effect could also be demonstrated in vesicles of phospholipids and was lost 4 days after withdrawal of the ethanol from the diet. These results closely parallel those relating to resistance to lipid disordering and suggest that both techniques detect a common modification. Lipid order was assessed using fluorescence anisotropy measurements of a range of fluorophores, including those used to determine the partitioning properties of the membrane. No effect of chronic ethanol treatment on lipid order was found, either in the intact membranes or in vesicles of extracted phospholipids. This suggests that changes in baseline order are not critical features of membrane tolerance in liver microsomes. In addition it appears that the altered partitioning of the 5-doxyl decane into the central region of the membrane is not related to lipid order changes in this region. The reduced partitioning of 5-doxyl decane may be a reflection of a redistribution in the lipid bilayer, perhaps due to modifications in other locations in the membrane, such as the lipid head group region.