Patergnani Simone, Pinton Paolo
Department of Morphology, Surgery and Experimental Medicine, Section of Pathology, Oncology and Experimental Biology, Laboratory for Technologies of Advanced Therapies (LTTA), University of Ferrara, Via Luigi Borsari, 46, Ferrara, 44121, Italy.
Methods Mol Biol. 2015;1241:181-94. doi: 10.1007/978-1-4939-1875-1_15.
Mitochondria are highly dynamic organelles, with a morphology ranging from small roundish elements to large interconnected networks. This fine architecture has a significant impact on mitochondrial homeostasis, and mitochondrial morphology is highly connected to specific cellular process. Autophagy is a catabolic process in which cell constituents, including proteins and organelles, are delivered to the lysosomal compartment for degradation. Autophagy has multiple physiological functions and recent advances have demonstrated that this process is linked to different human diseases, such as cancer and neurodegenerative disorders.In particular, it has been found that autophagy is a key determinant for the life span of mitochondria through a particularly fine-tuned mechanism called mitophagy, a selective form of autophagy, which ensures the preservation of healthy mitochondria through the removal of damaged or superfluous mitochondria. Mitophagy has been found to be altered in several pathologies and aberrant or excessive levels of this process are found in common human disorders. Thus, the measurement of the mitophagy levels is of fundamental relevance to elucidate the molecular mechanism of this process and, most importantly, its role in cellular homeostasis and disease.In this review, we will provide an overview of the current methods used to measure mitophagic levels, with particular emphasis on the techniques based on fluorescent probes.
线粒体是高度动态的细胞器，其形态范围从小的圆形结构到大型相互连接的网络。这种精细的结构对线粒体稳态有重大影响，并且线粒体形态与特定的细胞过程高度相关。自噬是一种分解代谢过程，其中包括蛋白质和细胞器在内的细胞成分被输送到溶酶体区室进行降解。自噬具有多种生理功能，最近的进展表明，这一过程与不同的人类疾病有关，如癌症和神经退行性疾病。特别是，已经发现自噬是线粒体寿命的关键决定因素，通过一种称为线粒体自噬的特别精细调节的机制，这是一种自噬的选择性形式，它通过清除受损或多余的线粒体来确保健康线粒体的保存。已经发现线粒体自噬在几种病理状态下会发生改变，并且在常见的人类疾病中发现该过程的异常或过度水平。因此，测量线粒体自噬水平对于阐明这一过程的分子机制至关重要，最重要的是，对于其在细胞稳态和疾病中的作用也至关重要。在这篇综述中，我们将概述目前用于测量线粒体自噬水平的方法，特别强调基于荧光探针的技术。
