Song Yutong, Huang Peiyuan, Duan Liting
Department of Biomedical Engineering, The Chinese University of Hong Kong, Hong Kong, China.
Methods Mol Biol. 2025;2840:185-200. doi: 10.1007/978-1-0716-4047-0_14.
Mitochondria are dynamic organelles with constantly changing morphologies. Despite recent reports indicating that mechanical cues modulate mitochondrial morphologies and functions, there is a lack of methods that can exclusively and precisely exert mechanical forces to and deform mitochondria in live cells. Therefore, how mitochondria sense and respond to mechanical forces remains largely elusive. Optogenetic methods open up new venues for remote and precise manipulation of intracellular activities using light, providing an unprecedented opportunity to establish targeted mechano-stimulation toward mitochondria. This chapter describes the development of a novel optogenetic approach to optically mechanostimulate and induce the deformation of mitochondria. In this approach, light-gated protein-protein heterodimerization recruits force-generating molecular motors to the outer mitochondrial membrane, enabling direct exertion of mechanical force on mitochondria. Details for the design, application, and experimental procedures are laid out in this chapter. This method presents a mitochondria-specific mechano-stimulator for studying the correlation between mitochondrial morphology and functions as well as mitochondrial mechanobiology.
线粒体是形态不断变化的动态细胞器。尽管最近有报道表明机械信号可调节线粒体的形态和功能，但缺乏能够在活细胞中专门且精确地对线粒体施加机械力并使其变形的方法。因此，线粒体如何感知和响应机械力在很大程度上仍然未知。光遗传学方法为利用光远程精确操纵细胞内活动开辟了新途径，为建立针对线粒体的靶向机械刺激提供了前所未有的机会。本章描述了一种新型光遗传学方法的开发，该方法可通过光学方式对线粒体进行机械刺激并诱导其变形。在这种方法中，光门控蛋白 - 蛋白异二聚化将产生力的分子马达招募到线粒体外膜，从而能够直接对线粒体施加机械力。本章详细介绍了该方法的设计、应用和实验步骤。该方法为研究线粒体形态与功能之间的相关性以及线粒体力学生物学提供了一种线粒体特异性机械刺激器。
