文章基本信息

标题：A comparative study on fluorescent cholesterol analogs as versatile cellular reporters
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作者：Erdinc Sezgin ; Fatma Betul Can ; Falk Schneider 等
期刊名称：JLR Papers In Press
印刷版ISSN：0022-2275
电子版ISSN：1539-7262
出版年度：2016
卷号：57
期号：2
页码：299-309
DOI：10.1194/jlr.M065326
语种：English
出版社：American Society for Biochemistry and Molecular Biology
摘要：Cholesterol (Chol) is a crucial component of cellular membranes, but knowledge of its intracellular dynamics is scarce. Thus, it is of utmost interest to develop tools for visualization of Chol organization and dynamics in cells and tissues. For this purpose, many studies make use of fluorescently labeled Chol analogs. Unfortunately, the introduction of the label may influence the characteristics of the analog, such as its localization, interaction, and trafficking in cells; hence, it is important to get knowledge of such bias. In this report, we compared different fluorescent lipid analogs for their performance in cellular assays: 1 ) plasma membrane incorporation, specifically the preference for more ordered membrane environments in phase-separated giant unilamellar vesicles and giant plasma membrane vesicles; 2 ) cellular trafficking, specifically subcellular localization in Niemann-Pick type C disease cells; and 3 ) applicability in fluorescence correlation spectroscopy (FCS)-based and super-resolution stimulated emission depletion-FCS-based measurements of membrane diffusion dynamics. The analogs exhibited strong differences, with some indicating positive performance in the membrane-based experiments and others in the intracellular trafficking assay. However, none showed positive performance in all assays. Our results constitute a concise guide for the careful use of fluorescent Chol analogs in visualizing cellular Chol dynamics.
关键词：cholesterol/trafficking ; fluorescence microscopy ; cholesterol/metabolism ; lipid rafts ; Niemann-Pick type C disease ; membranes/model ; giant unilamellar vesicles ; giant plasma membrane vesicles ; fluorescence correlation spectroscopy ; stimulated emission depletion