Systemic and strict regulation of the glutathione redox state in mitochondria and cytosol is needed for zebrafish ontogeny

• Published in: Biochimica et biophysica acta. General subjects Vol. 1868; no. 6; p. 130603
• Main Authors: Hamre, Kristin, Zhang, Wuxiao, Austgulen, Maren Hoff, Mykkeltvedt, Eva, Yin, Peng, Berntssen, Marc, Espe, Marit, Berndt, Carsten
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 01.06.2024
• Subjects: abnormal development; Animals; brain; cytosol; Cytosol - metabolism; Danio rerio; death; disulfides; Embryo, Nonmammalian - metabolism; Embryonic Development; gastrulation; glutathione; Glutathione - metabolism; Glutathione Disulfide - metabolism; green fluorescent protein; homeostasis; metabolism; mitochondria; Mitochondria - metabolism; muscles; ontogeny; organogenesis; oxidation; Oxidation-Reduction; oxidoreductases; perfluorooctane sulfonic acid; PFOS; pollutants; pollution; roGFP probe; Zebrafish - embryology; Zebrafish - metabolism; Zebrafish model; Embryonic development; GSSG; NAC; Zebrafish model; EhGSH; Glutathione metabolism; GSH; roGFP probe; PFOS; BSO; hpf
• DOI: 10.1016/j.bbagen.2024.130603
• ISSN: 0304-4165; 1872-8006

Redox control seems to be indispensable for proper embryonic development. The ratio between glutathione (GSH) and its oxidized disulfide (GSSG) is the most abundant cellular redox circuit. We used zebrafish harboring the glutaredoxin 1-redox sensitive green fluorescent protein (Grx1-roGFP) probe either in mitochondria or cytosol to test the hypothesis that the GSH:GSSG ratio is strictly regulated through zebrafish embryogenesis to sustain the different developmental processes of the embryo. Following the GSSG:GSH ratio as a proxy for the GSH-dependent reduction potential (EhGSH) revealed increasing mitochondrial and cytosolic EhGSH during cleavage and gastrulation. During organogenesis, cytosolic EhGSH decreased, while that of mitochondria remained high. The similarity between EhGSH in brain and muscle suggests a central regulation. Modulation of GSH metabolism had only modest effects on the GSSG:GSH ratios of newly hatched larvae. However, inhibition of GSH reductase directly after fertilization led to dead embryos already 10 h later. Exposure to the emerging environmental pollutant Perfluorooctane Sulfonate (PFOS) disturbed the apparent regulated EhGSH as well. Mitochondrial and cytosolic GSSG:GSH ratios are almost identical in different organs during zebrafish development indicating that the EhGSH might follow H2O2 levels and rather indirectly affect specific enzymatic activities needed for proper embryogenesis. Our data confirm that vertebrate embryogenesis depends on strictly regulated redox homeostasis. Disturbance of the GSSG:GSH circuit, e.g. induced by environmental pollution, leads to malformation and death. [Display omitted] •No organ specific mitochondrial and cytosolic GSH:GSSG ratios in zebrafish larvae.•Almost no effects of manipulated GSH metabolism on EhGSH in older zebrafish larvae.•Inhibition of GSH reductase in young larvae leads to malformation and death.•The environmental pollutant Perfluorooctane Sulfonate affects EhGSH in larvae.