ADH modulates plasma membrane lipid order of living MDCK cells via a cAMP-dependent process.
Using 1-[4-(trimethylamino)phenyl]-6-phenyl-hexa-1,3,5-triene, a fluorescent probe that specifically labels the external leaflet of the plasma membrane of living cells, we examined the effects of antidiuretic hormone (ADH) and various agents known to raise intracellular adenosine 3',5'-cyclic monophosphate (cAMP) on the physical state of the plasma membrane of Madin-Darby canine kidney (MDCK) cells. In polarized cells grown as a monolayer, [desamino-Cys1, DArg8]-vasopressin (V2-agonist) elicited a biphasic decrease in the lipid order as estimated from the decrease in fluorescence anisotropy (from r = 0.317 to r = 0.304, 37 degrees C) of the apical domain of the plasma membrane, equivalent at the peak response (t = 5 min) to that produced by an upward shift in temperature of 5-6 degrees C. A similar response was obtained by adding dibutyryl cAMP to the monolayers. Experiments on cell suspensions further indicated that the biphasic decrease in lipid order could also be evoked by forskolin, prostaglandin E2, and bradykinin but not by bradykinin plus indomethacin and was inhibitable by the protein kinases inhibitor compound H7. These data demonstrate that the lipid order of the plasma membrane of MDCK cells can be modulated in situ by cAMP-dependent processes probably involving protein kinase A activity, i.e., that membrane "fluidity" might act in the regulation of the cellular function of living epithelial cells. They provide a rationale for the changes in lipophilic solute permeability that accompany the increase in water permeability of target cells on ADH administration.
Giocondi
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MC
Institut National de la Santé et de la Recherche Médicale U 251, Department de Physiologie, Faculté de Medécine Xavier-Bichat, Paris, France.
Friedlander
G
G
Le Grimellec
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C
D019307:1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine; D000818:Animals; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism / Q000502:physiology / Q000648:ultrastructure; D000242:Cyclic AMP / Q000378:metabolism / Q000502:physiology*; D004285:Dogs; D007546:Isoquinolines / Q000494:pharmacology; D007668:Kidney / Q000166:cytology* / Q000378:metabolism / Q000502:physiology; D008560:Membrane Fluidity / Q000187:drug effects / Q000502:physiology; D008563:Membrane Lipids / Q000378:metabolism* / Q000502:physiology; D008854:Microscopy, Electron; D010879:Piperazines / Q000494:pharmacology; D011493:Protein Kinase C / Q000037:antagonists & inhibitors; D014667:Vasopressins / Q000502:physiology*
D007546:Isoquinolines; D008563:Membrane Lipids; D010879:Piperazines; D014667:Vasopressins; D019307:1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine; D000242:Cyclic AMP; D011493:Protein Kinase C