使用BODIPY FL鞘脂类似物研究小鼠胚胎干细胞中的鞘脂代谢。
Using BODIPY FL-Sphingolipid Analogs to Study Sphingolipid Metabolism in Mouse Embryonic Stem Cells.
作者信息
Fan Wei, Li Xiaoling
机构信息
Signal Transduction Laboratory, National Institute of Environmental Health Sciences, National Institutes of Health, Research Triangle Park, Triangle Park, North Carolina, United States.
出版信息
Bio Protoc. 2022 Nov 20;12(22). doi: 10.21769/BioProtoc.4555.
Sphingolipids are important structural components of cellular membranes. They also function as prominent signaling molecules to control a variety of cellular events, such as cell growth, differentiation, and apoptosis. Impaired sphingolipid metabolism, particularly defects in sphingolipid degradation, has been associated with many human diseases. Fluorescence sphingolipid analogs have been widely used as efficient probes to study sphingolipid metabolism and intracellular trafficking in living mammalian cells. Compared with nitrobenzoxadiazole fluorophores (NBD FL), the boron dipyrromethene difluoride fluorophores (BODIPY FL) have much higher absorptivity and fluorescence quantum. These features allow more intensive labeling of cells for fluorescence microscopy imaging and flow cytometry analysis. Here, we describe a protocol employing BODIPY FL-labeled sphingolipid analogs to elucidate sphingolipid internalization, trafficking, and endocytosis in mouse embryonic stem cells. eLife (2022), DOI: 10.7554/eLife.67452 Graphical abstract.
鞘脂是细胞膜的重要结构成分。它们还作为重要的信号分子发挥作用，以控制各种细胞事件，如细胞生长、分化和凋亡。鞘脂代谢受损，尤其是鞘脂降解缺陷，与许多人类疾病有关。荧光鞘脂类似物已被广泛用作研究活哺乳动物细胞中鞘脂代谢和细胞内运输的有效探针。与硝基苯并恶二唑荧光团（NBD FL）相比，二氟硼二吡咯荧光团（BODIPY FL）具有更高的吸收率和荧光量子产率。这些特性使得能够对细胞进行更强烈的标记，用于荧光显微镜成像和流式细胞术分析。在这里，我们描述了一种使用BODIPY FL标记的鞘脂类似物来阐明小鼠胚胎干细胞中鞘脂内化、运输和内吞作用的方案。《eLife》（2022年），DOI：10.7554/eLife.67452 图形摘要。
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