Mito-kaede photoactivation and chase experiment for mitophagy: optimizing flux measurement via fluid exchange system.

IF 2.2 4区工程技术 Q3 BIOCHEMICAL RESEARCH METHODS

BioTechniques Pub Date : 2024-09-11 DOI:10.1080/07366205.2024.2372955

Hiroyuki Morinaga,Yoh Sugawara,Yoshinori Kitagawa,Jingyuan Chen,Nobuo Yasuda,Hiroki Ogata,Yoshihiro Yamaguchi,Masao Kaneki,Joseph A Jeevendra Martyn,Shingo Yasuhara

引用次数: 0

Abstract

Modulating autophagy and mitophagy, vital cellular quality control systems, offer therapeutic potential for critical illnesses. However, limited drug screening options hinder progress. We present a novel assay using the photoswitchable fluorescent reporter, mito-Kaede, to quantify mitophagy flux. Mito-Kaede's superior UV-induced photoconversion and brightness post-conversion make it ideal for prolonged mitochondrial dynamics tracking. Its specificity in responding to mitophagy, confirmed by parkin-knockout cells, adds value. When coupled with a custom fluid exchange system, enabling efficient medium changes, precise mitophagy observations become feasible. This mitophagy assay, alongside our methodological insights, can decipher mitophagy's role in pathology and supports drug screening efforts.

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有丝分裂的光激活和追逐实验：通过流体交换系统优化通量测量。

自噬和有丝分裂是重要的细胞质量控制系统，调节这两个系统可为危重疾病提供治疗潜力。然而，有限的药物筛选方案阻碍了研究的进展。我们介绍了一种利用光开关荧光报告物 mito-Kaede 来量化有丝分裂通量的新型检测方法。Mito-Kaede 具有优异的紫外线诱导光电转换能力和转换后的亮度，因此非常适合用于长时间的线粒体动态跟踪。经 Parkin 基因敲除细胞证实，它对有丝分裂反应的特异性增加了其价值。如果与定制的液体交换系统配合使用，就能有效地更换培养基，从而实现对有丝分裂的精确观察。这种有丝分裂检测方法与我们的方法论见解相结合，可以破译有丝分裂在病理学中的作用，并为药物筛选工作提供支持。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

BioTechniques 工程技术-生化研究方法

CiteScore

4.40

自引率

0.00%

发文量

审稿时长

3.3 months

期刊介绍： BioTechniques is a peer-reviewed, open-access journal dedicated to publishing original laboratory methods, related technical and software tools, and methods-oriented review articles that are of broad interest to professional life scientists, as well as to scientists from other disciplines (e.g., chemistry, physics, computer science, plant and agricultural science and climate science) interested in life science applications for their technologies. Since 1983, BioTechniques has been a leading peer-reviewed journal for methods-related research. The journal considers: Reports describing innovative new methods, platforms and software, substantive modifications to existing methods, or innovative applications of existing methods, techniques & tools to new models or scientific questions Descriptions of technical tools that facilitate the design or performance of experiments or data analysis, such as software and simple laboratory devices Surveys of technical approaches related to broad fields of research Reviews discussing advancements in techniques and methods related to broad fields of research Letters to the Editor and Expert Opinions highlighting interesting observations or cautionary tales concerning experimental design, methodology or analysis.