Determination of plasma membrane fluidity with a fluorescent analogue of sphingomyelin by FRAP measurement using a standard confocal microscope
Determination of plasma membrane fluidity with a fluorescent analogue of sphingomyelin by FRAP measurement using a standard confocal microscope
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DOI：
10.1016/s1385-299x(03)00016-3
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发表时间：
2003-03-01
期刊：
影响因子：
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通讯作者：
Drouet, B
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文献类型：
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作者：
Klein, C;Pillot, T;Drouet, B
膜扰动效应已被描述为神经退行性过程，如阿尔茨海默病和朊病毒疾病。例如，与阿尔茨海默病有关的非纤维淀粉样肽（Abeta）可能通过膜扰动[9]发挥其毒性。膜组织可以通过其对脂质横向扩散的影响来评估，脂质横向扩散本身可以通过FRAP（光漂白后荧光恢复）来测量。我们使用这种技术来研究β对膜流动性的影响（pilot等人，正在准备的手稿）。我们在这里提出了一个简单的适应FRAP使用标准共聚焦激光扫描显微镜（CLSM）。作为测试实验，我们分析了鞘磷脂的荧光模拟物的横向扩散，并能够证明其在甲基- β -环糊精（cdx）诱导的胆固醇消耗时增加。(C) 2003 Elsevier Science B.V.版权所有
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.