Fig. 1: Mitochondrial content is reduced in axons of ADOA RGCs.
From: Inhibition of autophagy curtails visual loss in a model of autosomal dominant optic atrophy
a Representative z-projections of stacks of confocal images of mtRFP (red) in primary RGCs co-transfected with the indicated plasmids and after 24 h fixed and immunostained with β-tubulin III (βtub III, grey). Axons and somas were magnified in the bottom panels. EV, empty vector. Bars, 20 μm. b Quantification of mitochondrial length from four independent experiments as in (a) (EV, n = 86; Opa1, n = 113; Opa1K301A, n = 82; Opa1R905*, n = 71; Opa1Q297V, n = 114 cells). ****p < 0.0001 vs. EV in one-way ANOVA/Tukey’s test. c Quantification of mitochondrial content in axons from four independent experiments as in (a) (EV, n = 58; Opa1, n = 65; Opa1K301A, n = 69; Opa1R905*, n = 54; Opa1Q297V, n = 64 cells). *p = 0.018 Opa1, p = 0.027 Opa1Q297V vs. EV; ****p < 0.0001 vs. EV in one-way ANOVA/Bonferroni’s test. d Representative z-projections of stacks of confocal images acquired 24 h after transfection of primary RGCs co-transfected with mtRFP (red) and YFP-LC3 (green, autophagosome-LC3, auto-LC3) and the indicated plasmids. The cytoplasmic YFP-LC3 signal (cyto-LC3) is pseudocolored in grey for the sake of clarity. The region corresponding to the soma was magnified in the inset. Bars, 20 μm. e Quantification of soma mitochondria and autophagosome distribution towards the axonal hillock in 4 independent experiments as in (d) (EV, n = 65 mitochondria and 49 autophagosomes; Opa1, n = 69 mitochondria and autophagosomes; Opa1K301A, n = 58 mitochondria and 47 autophagosomes; Opa1R905*, n = 69 mitochondria and autophagosomes; Opa1Q297V, n = 70 mitochondria and autophagosomes) **p = 0.0023 Opa1K301A vs. EV; p = 0.0084 Opa1R905* vs. EV; ****p < 0.0001 vs. EV in one-way ANOVA/Tukey’s test. f Quantification of mitochondria and autophagosome co-localization in six independent experiments as in (d). (EV, n = 86; Opa1, n = 86; Opa1K301A, n = 101; Opa1R905*, n = 82; Opa1Q297V, n = 91 cells). *p = 0.0153 Opa1K301A, p = 0.043 Opa1R905* vs. EV in one-way ANOVA/Tukey’s test. In box plots, centre line represents mean, bounds of boxes SEM, whiskers the 10th–90th percentiles; each dot represents independent experiments. Source data are provided as a Source Data file.
