Mito-kaede photoactivation and chase experiment for mitophagy: optimizing flux measurement via fluid exchange system
| Autor: | Hiroyuki Morinaga, Yoh Sugawara, Yoshinori Kitagawa, Jingyuan Chen, Nobuo Yasuda, Hiroki Ogata, Yoshihiro Yamaguchi, Masao Kaneki, Joseph A Jeevendra Martyn, Shingo Yasuhara |
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| Jazyk: | angličtina |
| Rok vydání: | 2024 |
| Předmět: | |
| Zdroj: | BioTechniques, Vol 76, Iss 8, Pp 381-393 (2024) |
| Druh dokumentu: | article |
| ISSN: | 07366205 1940-9818 0736-6205 |
| DOI: | 10.1080/07366205.2024.2372955 |
| Popis: | 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. |
| Databáze: | Directory of Open Access Journals |
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