Asymmetry of plasma membrane lipid order in Madin-Darby Canine Kidney cells.
Fluorescence anisotropy experiments have been done to estimate, in situ, the lipid order of the plasma membrane of polarized Madin-Darby Canine Kidney cells (MDCK) grown on glass cover slips and labeled by 1-[4-(trimethylamino)phenyl]-6-phenylhexa-1,3,5-triene (TMA-DPH), a specific marker of the plasma membrane of living cells. Fluorescence microscopy, back-exchange, and quenching experiments indicated that TMA-DPH labeled the highly ordered (r greater than or equal to 0.32, 37 degrees C) apical domain of the plasma membrane of confluent monolayers. Opening of tight junctions or addition of the probe to cell suspensions resulted in a homogeneous distribution of TMA-DPH over the cell surface and in a marked decrease in anisotropy (0.27 less than or equal to r less than or equal to 0.29) that was due neither to a direct effect of Ca2+ on the probe nor to a change in fluorescence lifetime. Our data indicate that the apical domain, likely the external leaflet, of the plasma membrane of polarized MDCK cells is much more ordered than its basolateral counterpart.
D000818:Animals; D002118:Calcium / Q000494:pharmacology; D002460:Cell Line; D004161:Diphenylhexatriene / Q000031:analogs & derivatives; D004285:Dogs; D004492:Edetic Acid / Q000494:pharmacology; D005454:Fluorescence Polarization; D007668:Kidney / Q000166:cytology*; D008081:Liposomes; D008560:Membrane Fluidity; D008563:Membrane Lipids / Q000032:analysis*; D008856:Microscopy, Fluorescence
D008081:Liposomes; D008563:Membrane Lipids; D004161:Diphenylhexatriene; C033628:1-(4-(trimethylamino)phenyl)-6-phenylhexa-1,3,5-triene; D004492:Edetic Acid; D002118:Calcium