Fluorescence imaging detection of nanodomain redox signaling events at organellar contacts.
STAR Protoc
; 3(1): 101119, 2022 03 18.
Article
em En
| MEDLINE
| ID: mdl-35098166
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 (GSSGGSH) 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+. For complete details on the use and execution of this profile, please refer to Booth et al. (2016) and Booth et al. (2021).
Palavras-chave
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Organelas
/
Transdução de Sinais
/
Imagem Óptica
Tipo de estudo:
Diagnostic_studies
Limite:
Humans
Idioma:
En
Revista:
STAR Protoc
Ano de publicação:
2022
Tipo de documento:
Article