Differential rate of cholesterol efflux from the apical and basolateral membranes of MDCK cells.

Epithelial cells contain two distinct membrane surfaces, the apical and basolateral plasma membranes, which have different lipid and protein compositions. In order to assess the effect of the compositional differences of the apical and basolateral membranes on their ability to undergo cholesterol efflux, MDCK cells were radiolabeled with [3H]cholesterol and grown as a polarized monolayer on filter inserts, that separate the upper apical compartment from the lower basolateral compartment. The rate of cholesterol efflux from the basolateral membrane into media containing HDL in the basolateral compartment was 6.3%/h +/-0.7, whereas HDL-mediated efflux from the apical membrane was approximately 3-fold slower (1.9%/h +/-0.3). In contrast, Fu5AH cells, which do not form distinct polarized membrane domains, had a similar rate of HDL-mediated cholesterol efflux into the apical and basolateral compartments. Similar to HDL, other cholesterol acceptors, namely LDL, bovine serum albumin, and a lipid emulsion, also showed a decreased rate of cholesterol efflux from the apical membrane surface versus the basolateral membrane. Compared to the basolateral membrane, the apical membrane was also found to be more resistant to cholesterol oxidase treatment, to bind less HDL, and to take up less cholesterol from the medium. In conclusion, cholesterol efflux occurred less readily from the apical membrane than from the basolateral membrane for all types of acceptors tested. These results suggest that differences in the composition of the apical and basolateral membrane lead to a relative decrease in cholesterol desorption from the apical membrane and hence a reduced rate of cholesterol efflux.

[1]  J. Hoeg,et al.  Polarized secretion of apoA-I and apoA-II by transfected MDCK cells. , 1995, Journal of lipid research.

[2]  G. Meer,et al.  Chapter 21 - Lipid Polarity and Sorting in Epithelial Cells , 1994 .

[3]  J. Hoeg,et al.  Reverse Cholesterol Transport , 1994 .

[4]  M. Phillips,et al.  Effects of membrane lipids and -proteins and cytoskeletal proteins on the kinetics of cholesterol exchange between high density lipoprotein and human red blood cells, ghosts and microvesicles. , 1992, Biochimica et biophysica acta.

[5]  A. Hubbard Targeting of membrane and secretory proteins to the apical domain in epithelial cells. , 1991, Seminars in cell biology.

[6]  J. Slot,et al.  Subcellular localization of Forssman glycolipid in epithelial MDCK cells by immuno-electronmicroscopy after freeze-substitution , 1991, The Journal of cell biology.

[7]  A. Mendez,et al.  Protein kinase C as a mediator of high density lipoprotein receptor-dependent efflux of intracellular cholesterol. , 1991, The Journal of biological chemistry.

[8]  M. Phillips,et al.  Effects of membrane lipid composition on the kinetics of cholesterol exchange between lipoproteins and different species of red blood cells. , 1990, Biochimica et biophysica acta.

[9]  W. Young,et al.  Polarity of neutral glycolipids, gangliosides, and sulfated lipids in MDCK epithelial cells. , 1988, Journal of lipid research.

[10]  G van Meer,et al.  Lipid sorting in epithelial cells. , 1988, Biochemistry.

[11]  G van Meer,et al.  Sorting of sphingolipids in epithelial (Madin-Darby canine kidney) cells , 1987, The Journal of cell biology.

[12]  W. J. Johnson,et al.  Mechanisms and consequences of cellular cholesterol exchange and transfer. , 1987, Biochimica et biophysica acta.

[13]  G. Meer,et al.  The function of tight junctions in maintaining differences in lipid composition between the apical and the basolateral cell surface domains of MDCK cells. , 1986, The EMBO journal.

[14]  V. Schumaker,et al.  [6] Sequential flotation ultracentrifugation , 1986 .

[15]  J. Ordovás,et al.  Metabolism of apolipoproteins A-I, A-II, and A-IV. , 1986, Methods in enzymology.

[16]  J. Oram Receptor-mediated transport of cholesterol between cultured cells and high-density lipoproteins. , 1986, Methods in enzymology.

[17]  M. Lefevre,et al.  Metabolism of apolipoprotein A-IV. , 1984, Journal of lipid research.

[18]  M. Phillips,et al.  Role of the plasma membrane in the mechanism of cholesterol efflux from cells. , 1984, Biochimica et biophysica acta.

[19]  G. Meer,et al.  Viruses budding from either the apical or the basolateral plasma membrane domain of MDCK cells have unique phospholipid compositions. , 1982, The EMBO journal.

[20]  N. Simmons,et al.  Identification of two strains of MDCK cells which resemble separate nephron tubule segments. , 1981, Biochimica et biophysica acta.

[21]  R. Geyer,et al.  The effects of serum albumin and phospholipid on sterol excretion in tissue culture cells. , 1978, Biochimica et biophysica acta.