利用多种标记策略可视化线粒体DNA（mtDNA）核小体的分布和动态变化。

Visualize the Distribution and Dynamics of Mitochondrial DNA (mtDNA) Nucleoids with Multiple Labeling Strategies.

作者信息

Di Xiangjun, Qin Jinshan, Sun Yujie, Su Qian Peter

机构信息

School of Biomedical Engineering, Faculty of Engineering and Information Technology, University of Technology Sydney, Sydney, NSW, Australia.

State Key Laboratory of Membrane Biology, Biomedical Pioneer Innovation Center (BIOPIC), School of Life Sciences, Peking University, Beijing, China.

出版信息

Methods Mol Biol. 2023;2615:79-88. doi: 10.1007/978-1-0716-2922-2_6.

DOI:10.1007/978-1-0716-2922-2_6

PMID:36807785

Abstract

Mitochondrial DNA (mtDNA) encodes a variety of rRNAs, tRNAs, and respiratory chain complex proteins. The integrity of mtDNA supports the mitochondrial functions and plays an essential role in numerous physiological and pathological processes. Mutations in mtDNA cause metabolic diseases and aging. The mtDNA within the human cells are packaged into hundreds of nucleoids within the mitochondrial matrix. Knowledge of how the nucleoids are dynamically distributed and organized within mitochondria is key to understanding mtDNA structure and functions. Therefore, visualizing the distribution and dynamics of mtDNA within mitochondria is a powerful approach to gain insights into the regulation of mtDNA replication and transcription. In this chapter, we describe the methods of observing mtDNA and its replication with fluorescence microscopy in both fixed and live cells using different labeling strategies.

摘要

线粒体DNA（mtDNA）编码多种核糖体RNA、转运RNA和呼吸链复合蛋白。mtDNA的完整性支持线粒体功能，并在众多生理和病理过程中发挥重要作用。mtDNA突变会导致代谢疾病和衰老。人类细胞内的mtDNA被包装在线粒体基质中的数百个类核中。了解类核如何在线粒体内动态分布和组织是理解mtDNA结构和功能的关键。因此，可视化mtDNA在线粒体内的分布和动态是深入了解mtDNA复制和转录调控的有力方法。在本章中，我们描述了使用不同标记策略在固定细胞和活细胞中通过荧光显微镜观察mtDNA及其复制的方法。