In this article we used transgenic zebrafish lines that express compartment-specific isoforms of the roGFP2-Orp1 and Grx1-roGFP2 biosensors, described in Panieri et al (2017) [1], to test the contribute of the pentose phosphate pathway and of the glutathione biosynthesis in the antioxidant capacity of myocardial and endothelial cells in vivo. The transgenic zebrafish embryos were subdued to metabolic inhibition and subsequently challenged with H2O2 or the redox-cycling agent menadione to respectively mimic acute or chronic oxidative stress. Confocal time-lapse recordings were performed to follow the compartmentalized H2O2 and EGSH changes in the cardiovascular tissues of zebrafish embryos at 48 h post fertilization. After sequential excitation at 405 nm and 488 nm the emission was collected between 500-520 nm every 2 min for an overall duration of 60 min. The 405/488 nm ratio was normalized to the initial value obtained before oxidants addition and plotted over time. The analysis and the interpretation of the data can be found in the associated article [1].
Keywords: Cardiovascular system; Genetically-encoded sensors; Glutathione; Oxidative stress; Pentose phosphate pathway; Zebrafish.