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Abstract: P716 ABCA1 MODULATE CELLULAR MIGRATION AND MEMBRANE RUFFLING INDUCED BY FREE CHOLESTEROL IN MACROPHAGES FOAM CELLS
Poster - BASIC SCIENCE - Macrophages Abstract: P716 Citation: Atherosclerosis Supplement 2009, Vol. 10, Issue 2
ABCA1 MODULATE CELLULAR MIGRATION AND MEMBRANE RUFFLING INDUCED BY FREE CHOLESTEROL IN MACROPHAGES FOAM CELLS E Favari1, MP Adorni1, S Costa1, A Granata2, A Corsini2, N Ronda3, R Gatti4, F Bernini1 1
Pharm. Sci., University of Parma, Parma; 2Pharm. Sci., University of Milan, Milano; 3Clin. Med., University of Parma, Parma; 4Exp. Med., University of Parma, Parma
Increased membrane free cholesterol (FC) causes ruffling and inhibits cellular migration. Objective: Evaluate how the ABCA1 expression, that increases membrane FC, could modulate the plasma membrane organization and the cellular migration induced by cholesterolenrichment. Methods: Macrophages from wild type (WT) and ABCA1 knock out (ABCA1-KO) mice were loaded with acetylated LDL (AcLDL) and ACAT inhibitor in the presence or not of probucol, a specific ABCA1 inhibitor. We then evaluated membrane ruffling by confocal microscopy and cellular migration by chemotaxis assay to 10nM C5a. Results: In WT macrophages cholesterol-enrichment induces membrane ruffling and inhibits cellular migration by 68%. ABCA1-KO cells are less sensitive to cholesterol-induced membrane ruffling and cellular migration is inhibited by only 20%. In WT but not in ABCA1-KO cells probucol antagonizes the effect on membrane ruffling. Using a cell line that have stably knocked down ABCA1 expression (J774-ABCA1-KD), cholesterol-loading caused a decrease in cell migration by 42.3% compare to 79.3% in control cells. The migration is restored by apoA-I but not by HDL2 that do not promote the ABCA1-mediated efflux. Conclusion: In absence of a specific extracellular acceptor, ABCA1 promotes membrane ruffling and inhibits cellular migration by increasing membrane cholesterol. This effect is antagonized by probucol. Funding: Grant from the Italian Ministry of University and grant from Istituto Nazionale per le Ricerche Cardiovascolari.
ABCA1 MODULATE CELLULAR MIGRATION AND MEMBRANE RUFFLING INDUCED BY FREE CHOLESTEROL IN MACROPHAGES FOAM CELLS E Favari1, MP Adorni1, S Costa1, A Granata2, A Corsini2, N Ronda3, R Gatti4, F Bernini1 1
Pharm. Sci., University of Parma, Parma; 2Pharm. Sci., University of Milan, Milano; 3Clin. Med., University of Parma, Parma; 4Exp. Med., University of Parma, Parma
Increased membrane free cholesterol (FC) causes ruffling and inhibits cellular migration. Objective: Evaluate how the ABCA1 expression, that increases membrane FC, could modulate the plasma membrane organization and the cellular migration induced by cholesterolenrichment. Methods: Macrophages from wild type (WT) and ABCA1 knock out (ABCA1-KO) mice were loaded with acetylated LDL (AcLDL) and ACAT inhibitor in the presence or not of probucol, a specific ABCA1 inhibitor. We then evaluated membrane ruffling by confocal microscopy and cellular migration by chemotaxis assay to 10nM C5a. Results: In WT macrophages cholesterol-enrichment induces membrane ruffling and inhibits cellular migration by 68%. ABCA1-KO cells are less sensitive to cholesterol-induced membrane ruffling and cellular migration is inhibited by only 20%. In WT but not in ABCA1-KO cells probucol antagonizes the effect on membrane ruffling. Using a cell line that have stably knocked down ABCA1 expression (J774-ABCA1-KD), cholesterol-loading caused a decrease in cell migration by 42.3% compare to 79.3% in control cells. The migration is restored by apoA-I but not by HDL2 that do not promote the ABCA1-mediated efflux. Conclusion: In absence of a specific extracellular acceptor, ABCA1 promotes membrane ruffling and inhibits cellular migration by increasing membrane cholesterol. This effect is antagonized by probucol. Funding: Grant from the Italian Ministry of University and grant from Istituto Nazionale per le Ricerche Cardiovascolari.