The influence of microtubule integrity on plasma membrane fluidity in L929 cells.
The aim of this work was to examine the possible influence of the integrity of the microtubule network on the plasma membrane fluidity of L929 mouse fibroblasts. The L929 cell line was selected for the ease of culture and the stability of its characteristics. The cells were treated with colchicine, nocodazole and vinblastine, three microtubule-depolymerizing drugs, at various concentrations and for various times. Membrane fluidity was assessed from fluorescence depolarization measurements with the plasma membrane probe TMA-DPH. Each of the drugs induced a significant, dose-dependent decrease in fluorescence anisotropy. The effect levelled off (5-7% decrease) after approximately 90 min of treatment, and could be unambiguously interpreted as resulting from an increase in membrane fluidity. The cumulative action of the drugs did not significantly increase the effect. The effects of colchicine and nocodazole could be reversed by incubation in drug-free medium, but not that of vinblastine. The results are discussed in correlation with the kinetics of the three drugs interaction with tubulin or microtubules. It is concluded that the microtubule integrity contributed to the high plasma membrane lipidic order, but less than other factors, like the lipid composition and the cholesterol content.
Rémy-Kristensen
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Laboratoire de Pharmacologie et Physico-chimie, UMR CNRS 7034, Illkirch, France.
Duportail
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Coupin
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Kuhry
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D000818:Animals; D002460:Cell Line; D003078:Colchicine / Q000494:pharmacology*; D004161:Diphenylhexatriene / Q000031:analogs & derivatives* / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D005454:Fluorescence Polarization; D005456:Fluorescent Dyes / Q000494:pharmacology; D008560:Membrane Fluidity / Q000187:drug effects*; D051379:Mice; D008856:Microscopy, Fluorescence; D008870:Microtubules / Q000187:drug effects / Q000378:metabolism*; D015739:Nocodazole / Q000494:pharmacology*; D013997:Time Factors; D014747:Vinblastine / Q000494:pharmacology*
D005456:Fluorescent Dyes; D004161:Diphenylhexatriene; D014747:Vinblastine; C033628:1-(4-(trimethylamino)phenyl)-6-phenylhexa-1,3,5-triene; D015739:Nocodazole; D003078:Colchicine