Fluorescence imaging detection of nanodomain redox signaling events at organellar contacts

Authors

David M. Booth, Thomas Jefferson UniversityFollow
Péter Várnai, Semmelweis University
Suresh K Joseph, Thomas Jefferson UniversityFollow
György Hajnóczky, Thomas Jefferson UniversityFollow

Document Type

Article

Publication Date

3-18-2022

Comments

This article is the author’s final published version in STAR Protocols, Volume 3, Issue 1, March 2022, Article number 101119.

The published version is available at https://doi.org/10.1016/j.xpro.2021.101119. Copyright © Booth et al.

Abstract

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, H2O2, and Ca2+.

Recommended Citation

Booth, David M.; Várnai, Péter; Joseph, Suresh K; and Hajnóczky, György, "Fluorescence imaging detection of nanodomain redox signaling events at organellar contacts" (2022). Department of Pathology, Anatomy, and Cell Biology Faculty Papers. Paper 342.
https://jdc.jefferson.edu/pacbfp/342

Creative Commons License

This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 4.0 License.

PubMed ID

35098166

Language

English