Fig. 1

AAV.hSyn.mt-roGFP expresses in neuronal mitochondria and is functional in vivo.a Diagram of construct of AAV.hSyn.mt-roGFP.WPRE. b Validation of pAAV.hSyn.mt-roGFP in vivo. 4-mo-old C57Bl/6 mice were intracortically injected with pAAV.hSyn.mt-roGFP and exposed to either the oxidant DTDP or the reducing agent DTT. Field of view shows two-photon images of the expression of the AAV (green) in the cortex of the C57BL/6 J mice. Blood vessels are labelled with Texas Red (TR) Dextran 70,000 MW (red). Ratiometric roGFP imaging shows pseudocolor images according to the pseudocolor scale on the bottom, in basal conditions and after topical application of DTDP or DTT onto the brain surface. Scale bars represent 10 μm. c Scatter dot plot represents the Ratio 800/900 in neuronal mitochondria of every volume acquired. Error bars represent mean ± SEM. (Basal: 1.06 ± 0.036, DTT: 1.07 ± 0.044, DTDP: 1.55 ± 0.072; ****p < 0.0001, n = 33, 12 and 20 z-stacks from 7, 3 and 4 mice respectively, all male). d Scatter dot plot represents the relative change (%) in ratio for each condition. Error bars represent mean ± SEM (DTT: -0.96 ± 1.997%, n = 12 z-stacks; DTDP: 58.18 ± 8.719%, n = 20 z-stacks from 3 and 4 male mice respectively, ****p < 0.0001). e, f Histograms of Ratio 800/900 frequency distribution in basal conditions (grey) and 20 min after application of DTT 1 mM (e, purple) and DTDP 1 mM (f, orange) in neurons