Optogenetic Studies of Mitochondria.
Optogenetic Studies of Mitochondria.
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DOI：
10.1007/978-1-0716-2329-9_15
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发表时间：
2022
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虽然光遗传学方法已广泛用于远程控制细胞膜兴奋性和细胞内信号传导途径，但它们在线粒体研究中的应用受到限制，这主要是由于在线粒体中有效和特异性表达异源光门控视紫红质通道的挑战。在这里，我们描述了在线粒体内膜中表达功能性通道视紫红质2（ChR2）蛋白的方法，该蛋白具有异常长的线粒体前导序列，并表征了光遗传学介导的线粒体膜电位（Δ λ m）去极化。然后，我们说明了这种下一代光遗传学方法如何用于研究Δ m对线粒体功能的影响，如线粒体自噬，程序性细胞死亡和预处理介导的细胞保护。我们预计，这项创新技术将使新的见解的机制，其中的变化，在Δ λ m差异影响线粒体和细胞功能。
While optogenetic approaches have been widely used for remote control of cell membrane excitability and intracellular signaling pathways, their application in mitochondrial study has been limited, largely due to the challenge of effectively and specifically expressing heterologous light-gated rhodopsin channels in the mitochondria. Here, we describe the methods for expressing functional channelrhodopsin 2 (ChR2) proteins in the mitochondrial inner membrane with an unusually long mitochondrial leading sequence and characterizing optogenetic-mediated mitochondrial membrane potential (ΔΨm) depolarization. We then illustrate how this next-generation optogenetic approach can be used to study the effect of ΔΨm on mitochondrial functions such as mitophagy, programed cell death and preconditioning-mediated cytoprotection. We anticipate that this innovative technology will enable new insights into the mechanisms by which changes in ΔΨm differentially impacts mitochondrial and cellular functions.