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使用荧光标记对受体介导的自噬进行具有成本效益且简单的流式细胞术定量分析。

Cost-effective and simple flow cytometry quantification of receptor-mediated autophagy using fluorescent tagging.

作者信息

Marinković Mija, Rožić Ana, Polančec Denis, Novak Ivana

机构信息

Faculty of Science, University of Split, Croatia.

School of Medicine, University of Split, Croatia.

出版信息

FEBS Open Bio. 2025 Apr;15(4):587-598. doi: 10.1002/2211-5463.13958. Epub 2024 Dec 23.

DOI:10.1002/2211-5463.13958
PMID:39716041
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11961372/
Abstract

Mitophagy, a selective clearance of damaged or superfluous mitochondria via autophagy machinery and lysosomal degradation, is an evolutionarily conserved process essential for various physiological functions, including cellular differentiation and immune responses. Defects in mitophagy are implicated in numerous human diseases, such as neurodegenerative disorders, cancer, and metabolic conditions. Despite significant advancements in mitophagy research over recent decades, novel and robust methodologies are necessary to elucidate its molecular mechanisms comprehensively. In this study, we present a detailed protocol for quantitatively assessing mitophagy through flow cytometry using a mitochondria-targeted fluorescent mitophagy receptor, GFP-BNIP3L/NIX. This method offers a rapid alternative to conventional microscopy or immunoblotting techniques for analyzing mitophagy activity. Additionally, this approach can theoretically be adapted to utilize any fluorescent-tagged selective autophagy receptor, enabling the direct and rapid analysis of various types of receptor-mediated selective autophagy.

摘要

线粒体自噬是通过自噬机制和溶酶体降解对受损或多余线粒体进行选择性清除的过程，是一种进化上保守的过程，对包括细胞分化和免疫反应在内的各种生理功能至关重要。线粒体自噬缺陷与许多人类疾病有关，如神经退行性疾病、癌症和代谢性疾病。尽管近几十年来线粒体自噬研究取得了重大进展，但仍需要新颖且强大的方法来全面阐明其分子机制。在本研究中，我们展示了一种详细的方案，该方案使用线粒体靶向荧光线粒体自噬受体GFP-BNIP3L/NIX，通过流式细胞术对线粒体自噬进行定量评估。该方法为分析线粒体自噬活性提供了一种快速替代传统显微镜或免疫印迹技术的方法。此外，从理论上讲，这种方法可以适用于利用任何荧光标记的选择性自噬受体，从而能够直接快速地分析各种类型的受体介导的选择性自噬。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a838/11961372/886da123140d/FEB4-15-587-g010.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a838/11961372/5df77304f21d/FEB4-15-587-g008.jpg
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https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a838/11961372/493564b5d0f9/FEB4-15-587-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a838/11961372/56a98389d481/FEB4-15-587-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a838/11961372/66c55ca1f65e/FEB4-15-587-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a838/11961372/886da123140d/FEB4-15-587-g010.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a838/11961372/5df77304f21d/FEB4-15-587-g008.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a838/11961372/932541fe7ab0/FEB4-15-587-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a838/11961372/76cafbb76a47/FEB4-15-587-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a838/11961372/7a4140b6e592/FEB4-15-587-g007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a838/11961372/3d8c5d63bb35/FEB4-15-587-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a838/11961372/493564b5d0f9/FEB4-15-587-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a838/11961372/56a98389d481/FEB4-15-587-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a838/11961372/66c55ca1f65e/FEB4-15-587-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a838/11961372/886da123140d/FEB4-15-587-g010.jpg

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2
Autophagy in health and disease: From molecular mechanisms to therapeutic target.健康与疾病中的自噬：从分子机制到治疗靶点
MedComm (2020). 2022 Jul 10;3(3):e150. doi: 10.1002/mco2.150. eCollection 2022 Sep.
3
Guidelines for the use and interpretation of assays for monitoring autophagy (4th edition).
自噬监测分析方法使用和解释的指南（第 4 版）。
Autophagy. 2021 Jan;17(1):1-382. doi: 10.1080/15548627.2020.1797280. Epub 2021 Feb 8.
4
The autophagy research in electron microscopy.电子显微镜下的自噬研究
Appl Microsc. 2019 Nov 6;49(1):11. doi: 10.1186/s42649-019-0012-6.
5
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6
A Mammalian Mitophagy Receptor, Bcl2-L-13, Recruits the ULK1 Complex to Induce Mitophagy.一种哺乳动物的自噬受体 Bcl2-L-13 招募 ULK1 复合物诱导自噬。
Cell Rep. 2019 Jan 8;26(2):338-345.e6. doi: 10.1016/j.celrep.2018.12.050.
7
Phosphorylation of the mitochondrial autophagy receptor Nix enhances its interaction with LC3 proteins.线粒体自噬受体 Nix 的磷酸化增强了它与 LC3 蛋白的相互作用。
Sci Rep. 2017 Apr 25;7(1):1131. doi: 10.1038/s41598-017-01258-6.
8
FKBP8 recruits LC3A to mediate Parkin-independent mitophagy.FKBP8招募LC3A以介导不依赖帕金蛋白的线粒体自噬。
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10
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Cell Mol Life Sci. 2016 Feb;73(4):775-95. doi: 10.1007/s00018-015-2087-8. Epub 2015 Nov 26.