Fig. 6

Mitochondrial candidates modulate synaptic morphology in vivo. a. Representative image of WT muscle 12 NMJ in abdominal segment A3 (n = 8) immunostained with anti-HRP. b-e. Top panel displays representative images of muscle 12 NMJ, hemisegment A3 bouton morphology with candidate knock-down. Middle panel demonstrates masks utilized for quantification in ImageJ. Lower panel shows a section of axon. Scale bar = 10um for synaptic HRP and mask layers, 5um for axons. b. elav-GAL4/Ndufb8RNAi; c. elav-GAL4/MitofilinRNAi; d. elav-GAL4/Vdac1RNAi; e. elav-GAL4/AldhRNAi. Arrows represent loss or alteration of boutons and/or fragmentation of dendritic branches. A variety of phenotypes exist. All NMJs immunostained with anti-HRP and imaged at 63× (n = 8). f. Quantification of axon diameter in candidate lines versus control (Control mean ± SEM = 1.88 ± 0.02; Ndufb8 p=0.4095, mean ± SEM = 2.19 ± 0.26; Mitofilin p=0.169, mean ± SEM = 1.52 ± 0.05; Vdac1 p=0.1427, mean ± SEM = 1.52 ± 0.15; Aldh p=0.3138, mean ± SEM = 2.28 ± 0.26). g. Quantification of number of NMJ branches in candidate lines versus control (Control mean ± SEM = 6.0 ± 0.0; Ndufb8 p=0.1161, mean ± SEM = 6.66 ± 0.33; Mitofilin p=0.117, mean ± SEM = 5.33 ± 0.33; Vdac1 p=0.2722, mean ± SEM = 5.5 ± 0.5; Aldh p=0.0522, mean ± SEM = 5.0 ± 0.0). h. Quantification of the synaptic bouton diameter in candidate lines versus control (Control mean ± SEM = 1.95 ± 0.06; Ndufb8 p=0.0003, mean ± SEM = 2.43 ± 0.1; Mitofilin p=0.5759, mean ± SEM = 2.02 ± 0.09; Vdac1 p=0.0003, mean ± SEM = 2.51 ± 0.15; Aldh p=0.5291, mean ± SEM = 1.87 ± 0.1). i. Quantification of the total bouton area in candidate versus control lines (Control mean ± SEM = 5.08 ± 0.47; Ndufb8 p=0.4246, mean ± SEM = 4.57 ± 0.44; Mitofilin p=0.0078, mean ± SEM = 3.3 ± 0.23; Vdac1 p=0.6083, mean ± SEM = 4.68 ± 0.47; Aldh p=0.0337, mean ± SEM = 3.6 ± 0.46). All quantification used students t-test; * = p<0.05, ** = p<0.01, *** = p<0.001, **** = p<0.0001