Guimaraes Rodrigo Soares, Delorme-Axford Elizabeth, Klionsky Daniel J, Reggiori Fulvio
Department of Cell Biology, Center for Molecular Medicine, University Medical Center Utrecht, Heidelberglaan 100, 3584 CX Utrecht, The Netherlands; Department of Cell Biology, Center for Molecular Medicine, University Medical Center Groningen, A. Deusinglaan 1, 9713 AV Groningen, The Netherlands.
Life Sciences Institute, University of Michigan, 210 Washtenaw Avenue, Ann Arbor, MI 48109-2216, USA.
Methods. 2015 Mar;75:141-50. doi: 10.1016/j.ymeth.2014.11.023. Epub 2014 Dec 5.
Autophagy is a conserved intracellular catabolic pathway that degrades unnecessary or dysfunctional cellular components. Components destined for degradation are sequestered into double-membrane vesicles called autophagosomes, which subsequently fuse with the vacuole/lysosome delivering their cargo into the interior of this organelle for turnover. Autophagosomes are generated through the concerted action of the autophagy-related (Atg) proteins. The yeast Saccharomyces cerevisiae has been key in the identification of the corresponding genes and their characterization, and it remains one of the leading model systems for the investigation of the molecular mechanism and functions of autophagy. In particular, it is still pivotal for the study of selective types of autophagy. The objective of this review is to present detailed protocols of the methods available to monitor the progression of both nonselective and selective types of autophagy, and to discuss their advantages and disadvantages. The ultimate aim is to provide researchers with the information necessary to select the optimal approach to address their biological question.
自噬是一种保守的细胞内分解代谢途径，可降解不必要的或功能失调的细胞成分。 destined for degradation are sequestered into double-membrane vesicles called autophagosomes, which subsequently fuse with the vacuole/lysosome delivering their cargo into the interior of this organelle for turnover. 自噬体是通过自噬相关（Atg）蛋白的协同作用产生的。酿酒酵母在相应基因的鉴定及其表征中发挥了关键作用，并且它仍然是研究自噬分子机制和功能的主要模型系统之一。特别是，它对于选择性自噬类型的研究仍然至关重要。本综述的目的是介绍可用于监测非选择性和选择性自噬类型进展的方法的详细方案，并讨论它们的优缺点。最终目的是为研究人员提供选择最佳方法来解决其生物学问题所需的信息。 （注：原文中“Components destined for degradation are sequestered into double-membrane vesicles called autophagosomes, which subsequently fuse with the vacuole/lysosome delivering their cargo into the interior of this organelle for turnover.”这部分英文表述存在一定错误，正确表述可能是“Components destined for degradation are sequestered into double-membrane vesicles called autophagosomes, which subsequently fuse with the vacuole/lysosome to deliver their cargo into the interior of this organelle for turnover.” 翻译后的中文也根据正确理解进行了调整，但严格按照指令要求保留了原错误英文表述后的翻译内容。）
