Fluorescence imaging detection of nanodomain redox signaling events at organellar contacts.
| Autor: | Booth DM; MitoCare Center, Department of Pathology, Anatomy and Cell Biology, Thomas Jefferson University, Jefferson Alumni Hall 1020 Locust Street, Philadelphia, PA 19107, USA., Várnai P; Department of Physiology, Semmelweis University, Faculty of Medicine, 1444 Budapest, Hungary., Joseph SK; MitoCare Center, Department of Pathology, Anatomy and Cell Biology, Thomas Jefferson University, Jefferson Alumni Hall 1020 Locust Street, Philadelphia, PA 19107, USA., Hajnóczky G; MitoCare Center, Department of Pathology, Anatomy and Cell Biology, Thomas Jefferson University, Jefferson Alumni Hall 1020 Locust Street, Philadelphia, PA 19107, USA. |
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| Jazyk: | angličtina |
| Zdroj: | STAR protocols [STAR Protoc] 2022 Jan 20; Vol. 3 (1), pp. 101119. Date of Electronic Publication: 2022 Jan 20 (Print Publication: 2022). |
| DOI: | 10.1016/j.xpro.2021.101119 |
| Abstrakt: | This protocol describes how to visualize, detect, and analyze redox signals (oxidative bursts) at the ER-mitochondrial interface. It uses drug-inducible crosslinking to target the genetically encoded glutathione redox sensor Grx1roGFP2 to organellar contact sites to measure local redox changes associated with transient depolarizations of the mitochondrial membrane potential (flickers). The strategy allows imaging of the oxidized to reduced glutathione ratio (GSSG:GSH) in subcellular regions below the diffraction limit with good temporal resolution and minimum phototoxicity. Moreover, the strategy also applies to diverse parameters including pH, H Competing Interests: The authors declare no competing interests. (© 2022 The Authors.) |
| Databáze: | MEDLINE |
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