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Fig. 1 | Acta Neuropathologica Communications

Fig. 1

From: In vivo real-time dynamics of ATP and ROS production in axonal mitochondria show decoupling in mouse models of peripheral neuropathies

Fig. 1

In vivo validation of the fluorescent probes. a Several axons expressing mito-roGFP-Orp1 or mito-ATeam probe can be observed in teased fibers of mouse sciatic nerve 1 month after the virus injection. CARS imaging (inserts) shows that axons expressing mito-roGFP-Orp1 or mito-ATeam (green) are surrounded by a myelin sheath (red). Scale bars = 3 μm. b Mito-roGFP-Orp1 expression (green) partially colocalizes with mitochondrial marker TOM20 (red) in the axon. Similarly, mito-ATeam partially colocalizes with TOM20 in axonal mitochondria. This partial colocalization is due to the heterogenous distribution of TOM20 in outer mitochondrial membrane [74]. Scale bars = 3 μm. c A fluorescent signal of mito-roGFP-Orp1 is detected when the probe is reduced and when it is oxidized by H2O2. The ratio of mito-roGFP-Orp1 fluorescence measured in vivo (n = 6 axons; 6 mice) shows a slight increase over time. Scale bar = 10 μm d Mito-roGFP-Orp1 fluorescence ratio decreases with DTT (p = 0.006; n = 3 axons, 3 mice) and increases after injection of diamide into the nerve (p = 0.003; n = 4 axons, 4 mice). e A fluorescent signal of the CFP subunit and Venus subunit of mito-ATeam is detected. Mito-ATeam fluorescence ratio measured in vivo shows no significant change over time (n = 18 axons; 3 mice). Scale bar = 10 μm. f Mito-ATeam fluorescence ratio increases after injection of 0.4 M (p = 0.06; n = 3 axons, 3 mice) and 1 M (p = 7.5E-4; n = 6 axons, 4 mice) ATP into the sciatic nerve. All error bars show SEM. Statistical analysis shows Student two-tailed T-tests

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