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Papers In Press, published online ahead of print August 1, 2003
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Department of Medical Chemistry, Biochemistry and Biotechnology, University of Milano, Segrate, Milano 20090
Corresponding Author: alessandro.prinetti{at}unimi.it
ABSTRACT Treatments with methyl-
Revised on July 22, 2003
Accepted on July 23, 2003
Dynamics of membrane lipid domains in neuronal cells differentiated in culture
-cyclodextrin induced a time- and dose-dependent efflux of cholesterol, sphingolipids and phosphatidylcholine from cerebellar neurons differentiated in culture. In the case of a “mild” treatment, the loss of cell lipids induced a deep reorganization of the remaining membrane lipids. In fact, the amount of phosphatidylcholine associated with a Triton X-100-insoluble membrane fraction, (highly enriched in sphingolipids and cholesterol in non-treated cells) was lowered by the treatment, this suggesting a reduction of the lipid domain area. However, the cholesterol and sphingolipid enrichment of this fraction remained substantially unchanged, suggesting the existence of dynamic processes aimed to preserve the segregation of cholesterol and sphingolipids in membrane domains. Under these conditions, the lipid membrane domains retained their property to sort signaling proteins such as Lyn and c-Src, but cells displayed deep alterations in their membrane permeability. However, normal membrane permeability was restored by loading cells with cholesterol. When methyl--cyclodextrin treatment was more stringent, a large loss of cell lipids occurred, the lipid domains were much less enriched in cholesterol and lost the property to sort specific proteins. The loss of the integrity and properties of lipid domains was accompanied by severe changes in the membrane permeability, sufferance and eventually cell death.
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