Determination of plasma membrane fluidity with a fluorescent analogue of sphingomyelin by FRAP measurement using a standard confocal microscope

被引:30
作者
Klein, C
Pillot, T
Chambaz, J
Drouet, B
机构
[1] Inst Biomed Cordeliers, INSERM, IFR 58, Serv Commun Imagerie & Cytometrie, F-75006 Paris, France
[2] Univ Nancy 1, INSERM, E0014, F-54000 Nancy, France
[3] Univ Paris 06, INSERM, U505, F-75006 Paris, France
来源
BRAIN RESEARCH PROTOCOLS | 2003年 / 11卷 / 01期
关键词
plasma membrane; fluidity; lateral diffusion; FRAP; CLSM; sphingomyelin; neuroblastoma cells;
D O I
10.1016/S1385-299X(03)00016-3
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Membrane perturbing effects have been described in neurodegenerative process like Alzheimer's disease and prion disorders. For example, non fibrillar amyloid-beta peptides (Abeta) implicated in Alzheimer's disease may exert its toxicity via membrane perturbation [9]. Membrane organisation can be evaluated by its influence on lateral diffusion of lipids, which itself can be measured by FRAP (fluorescence recovery after photobleaching). We used this technique to study the effects of Abeta on membrane fluidity (Pillot et al., manuscript in preparation). We propose here a simple adaptation of FRAP using standard confocal laser scanning microscopy (CLSM). As a test experiment, we analysed the lateral diffusion of a fluorescent analogue of sphingomyelin and were able to demonstrate its increase upon cholesterol depletion induced by methyl-beta-cyclodextrin (cdx). (C) 2003 Elsevier Science B.V. All rights reserved.
引用
收藏
页码:46 / 51
页数:6
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