Guidelines for monitoring autophagy in Caenorhabditis elegans.
Guidelines for monitoring autophagy in Caenorhabditis elegans.
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DOI：
10.1080/15548627.2014.1003478
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发表时间：
2015
期刊：
影响因子：
13.3
通讯作者：
Wu F
中科院分区：
文献类型：
--
作者：
Zhang H;Chang JT;Guo B;Hansen M;Jia K;Kovács AL;Kumsta C;Lapierre LR;Legouis R;Lin L;Lu Q;Meléndez A;O'Rourke EJ;Sato K;Sato M;Wang X;Wu F
细胞自噬的循环过程已广泛表征与酵母和哺乳动物细胞系的标准测定。在多细胞生物中，在有机体个体发生的各个阶段，许多外部和内部因素对特定细胞类型的自噬活性有不同的影响，这增加了分析这些后生动物自噬的复杂性。在这里，我们总结了目前可用的检测方法来监测线虫的自噬过程。结合测量脂化at8同源物LGG-1的水平，降解特性良好的自噬底物，如种系P颗粒成分和SQSTM1/p62同源物SQST-1，自噬基因的表达和自噬结构的电镜分析，是目前可用来评估线虫自噬水平的最可靠的方法。我们还回顾了自噬活性的改变如何影响秀丽隐杆线虫的各种生物过程，如L1在饥饿条件下的存活、水的形成、衰老和细胞死亡，以及神经元细胞的特异性。综上所述，秀丽隐杆线虫正在成为一种强大的模式生物来监测自噬，同时评估自噬在关键发育事件和成年期的重要生理作用。
The cellular recycling process of autophagy has been extensively characterized with standard assays in yeast and mammalian cell lines. In multicellular organisms, numerous external and internal factors differentially affect autophagy activity in specific cell types throughout the stages of organismal ontogeny, adding complexity to the analysis of autophagy in these metazoans. Here we summarize currently available assays for monitoring the autophagic process in the nematode C. elegans. A combination of measuring levels of the lipidated Atg8 ortholog LGG-1, degradation of well-characterized autophagic substrates such as germline P granule components and the SQSTM1/p62 ortholog SQST-1, expression of autophagic genes and electron microscopy analysis of autophagic structures are presently the most informative, yet steady-state, approaches available to assess autophagy levels in C. elegans. We also review how altered autophagy activity affects a variety of biological processes in C. elegans such as L1 survival under starvation conditions, dauer formation, aging, and cell death, as well as neuronal cell specification. Taken together, C. elegans is emerging as a powerful model organism to monitor autophagy while evaluating important physiological roles for autophagy in key developmental events as well as during adulthood.