生物学杂志

• 特约综述 •    下一篇

细胞自噬研究方法的比较与分析

  

  1. 中国科学院 合肥物质科学研究院 强磁场科学中心, 合肥 230031
  • 出版日期:2018-08-18 发布日期:2018-08-18
  • 通讯作者: 张欣,博士,研究员,研究方向为细胞生物学与磁生物效应,E-mail: xinzhang@hmfl.ac.cn
  • 作者简介:李志元,博士,博士后,研究方向为自噬与细胞周期调控,E-mail: lizhiyuan@hmfl.ac.cn
  • 基金资助:
    中国科学院百人计划;中国博士后科学基金第60批面上项目资助(2016M602045)

Comparison and analysis of methods in autophagy research

  1. High Magnetic Field Laboratory, Hefei Institutes of Physical Science,Chinese Academy of Sciences, Hefei 230031, China
  • Online:2018-08-18 Published:2018-08-18

摘要: 自噬是一种在真核细胞内保守的降解过程,从单细胞的酵母到哺乳动物中都普遍存在。自噬通过降解细胞内受损的蛋白、细胞器、细胞质以及入侵的微生物等来维持机体的稳态。结合已有的文献报道和实验室的经验积累,总结了目前在细胞自噬研究中常用的一些研究方法:LC3 turnover、GFP的抗降解性、p62、mRFP-GFP-LC3活细胞成像、电镜和流式细胞仪检测等,以及在实验中需要注意的具体事项,如一些可以直接影响实验结果但并没有引起人们重视的因素。希望能为新进入细胞自噬领域的研究人员提供一些实验操作上的指导和参考,从而可以在自噬研究中得到真实准确及可重复的实验结果与合理的理论解释。

关键词: 自噬, 自噬潮, LC3, 自噬抑制剂

Abstract: Autophagy is an evolutionarily conserved degradation process in eukaryotes from yeast to mammalian cells, which degrades damaged proteins, organelles, cytosol and invaded microorganism to maintain homeostasis. Based on data reported in the literature and our own experience, we summarize some commonly used methods in autophagy research, such as LC3 turnover, GFP resistance to degradation, p62, mRFP-GFP-LC3 live imaging, electron microscope and flow cytometry. We also provide some experimental tips and details that people should pay attention to because these factors frequently cause experimental variations. We hope to provide some experimental guidance and references for researchers that are new to the field of autophagy. This will help people to get accurate and reproducible experimental results as well as valid explanations. 

Key words: autophagy, autophagic flux, LC3, autophagy inhibitor