Papandreou M E, Tavernarakis N
Institute of Molecular Biology and Biotechnology, Foundation for Research and Technology-Hellas, Heraklion, Greece; Faculty of Medicine, University of Crete, Heraklion, Greece.
Institute of Molecular Biology and Biotechnology, Foundation for Research and Technology-Hellas, Heraklion, Greece; Faculty of Medicine, University of Crete, Heraklion, Greece.
Methods Enzymol. 2017;588:429-444. doi: 10.1016/bs.mie.2016.10.039. Epub 2016 Dec 5.
Autophagy, from the Greek auto (self) and phagy (eating), is a self-degradative process critical for eukaryotic cell homeostasis. Its rapidly responsive, highly dynamic nature renders this process essential for adapting to and offsetting acute/harsh conditions such as starvation, organelle dysfunction, and deoxyribonucleic acid (DNA) damage. Autophagy involves an intricate network of interacting factors with multiple levels of control. Importantly, dysregulation of autophagy has been linked to numerous debilitating pathologies, including cancer and neurodegenerative conditions in humans. Methods to monitor and quantify autophagic activity reliably are essential both for studying the basic mechanisms of autophagy and for dissecting its involvement in disease. The nematode Caenorhabditis elegans is a particularly suitable model organism to effectively visualize and study autophagy, in vivo, in a physiological and pathological context due to its optical transparency, experimental malleability, and precise developmental and anatomical characterization. Here, we outline the main tools and approaches to monitor and measure autophagic responses in C. elegans.
自噬,源于希腊语auto(自我)和phagy(吞噬),是一种对真核细胞内稳态至关重要的自我降解过程。其快速响应、高度动态的特性使该过程对于适应和抵消诸如饥饿、细胞器功能障碍及脱氧核糖核酸(DNA)损伤等急性/恶劣条件至关重要。自噬涉及一个由相互作用因子构成的复杂网络,具有多个控制层面。重要的是,自噬失调与众多使人衰弱的病症相关,包括人类的癌症和神经退行性疾病。可靠地监测和量化自噬活性的方法对于研究自噬的基本机制以及剖析其在疾病中的作用都至关重要。线虫秀丽隐杆线虫因其光学透明性、实验可塑性以及精确的发育和解剖特征,是一种特别适合在生理和病理背景下在体内有效可视化和研究自噬的模式生物。在此,我们概述了监测和测量秀丽隐杆线虫自噬反应的主要工具和方法。