Assays for Mitophagy in Yeast
Assays for Mitophagy in Yeast
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DOI：
10.1007/978-1-4939-6824-4_20
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发表时间：
2017-01-01
期刊：
影响因子：
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通讯作者：
Okamoto, Koji
中科院分区：
文献类型：
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作者：
Eiyama, Akinori;Okamoto, Koji
消除受损或多余的线粒体对于维持细胞完整性和能量供需平衡至关重要。线粒体自噬以依赖于自噬的方式选择性地分解代谢线粒体，并有助于线粒体的质量和数量控制。该降解系统在包括芽殖酵母（Saccharomyces cerevisiae）在内的真核生物中高度保守。因此，使用酵母的线粒体自噬分析有可能提供深入了解线粒体自噬的常见机制。在这里，我们介绍了实验方法，荧光显微镜和蛋白质印迹法，以验证在酵母线粒体自噬。前者是有用的可视化运输线粒体的液泡，裂解室，并形成含自噬体。后者使我们能够量化线粒体降解。
Elimination of damaged or surplus mitochondria is crucial to maintain cellular integrity and an energy supply-demand balance. Mitophagy serves to selectively catabolize mitochondria in a manner dependent on autophagy, and contributes to mitochondrial quality and quantity control. This degradation system is highly conserved among eukaryotes including the budding yeast, Saccharomyces cerevisiae. Therefore, analyses of mitophagy using yeast have the potential to provide insights into the common mechanisms of mitophagy. Here, we introduce experimental approaches with fluorescence microscopy and western blotting to validate mitophagy in yeast. The former is useful to visualize transport of mitochondria to the vacuole, a lytic compartment, and formation of mitochondria-containing autophagosomes. The latter allows us to quantify mitochondria degradation.