In Vivo Mapping of Hydrogen Peroxide and Oxidized Glutathione Reveals Chemical and Regional Specificity of Redox Homeostasis

The glutathione redox couple (GSH/GSSG) and hydrogen peroxide (H2O2) are central to redox homeostasis and redox signaling, yet their distribution within an organism is difficult to measure. Using genetically encoded redox probes in Drosophila, we establish quantitative in vivo mapping of the glutathione redox potential (EGSH) and H2O2 in defined subcellular compartments (cytosol and mitochondria) across the whole animal during development and aging. A chemical strategy to trap the in vivo redox state of the transgenic biosensor during specimen dissection and fixation expands the scope of fluorescence redox imaging to include the deep tissues of the adult fly. We find that development and aging are associated with redox changes that are distinctly redox couple-, subcellular compartment-, and tissue-specific. Midgut enterocytes are identified as prominent sites of age-dependent cytosolic H2O2 accumulation. A longer life span correlated with increased formation of oxidants in the gut, rather than a decrease. ▸ Genetically encoded redox probes visualize redox differences and changes in vivo ▸ A chemical trapping strategy enables robust redox imaging in dissected animals ▸ Physiological redox changes are redox couple, compartment, and tissue specific ▸ Increased life span coincided with increased rather than decreased tissue oxidation