Neuronal stimulation and muscle contractions do not mediate mitochondrial patterning. (A) Confocal imaging of mitochondrial network (Tomm20, T20; green) counterstained with a marker for neuromuscular junctions αBungarotoxin (αBung; red) and a marker for primary motor neurons (ZNP1; magenta) at 24 and 48 hpf. While T20 signal corresponds to a unique confocal stack, αBung and ZNP1 represent the Z project. (B) Confocal imaging of mitochondrial network (Tomm20, T20; green) counterstained with αBungarotoxin (αBung; red) and primary motor neurons (ZNP1; magenta) at 28 hpf, with or without intravenous injection of αBungarotoxin performed at 24 hpf. (C) Confocal imaging of mitochondrial network (Tomm20, T20; green) counterstained with Phalloidin (Ph; red) at 24 hpf, with or without electrical pulse stimulation (EPS) applied at 20 hpf. (D) Confocal imaging of mitochondrial network (Tomm20, T20; green) counterstained with Phalloidin (Phall; red) at 28 hpf, with or without electrical pulse stimulation (EPS) applied at 24 hpf. (E) Quantification of Tomm20-zsGreen fluorescence ratio between somite center and boundary region at 24 and 28 hpf after 4 h of EPS (n = 6 fish per group, 3 images analyzed per fish).
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