Fig. 6: Analysis of mitochondria in E18 hippocampal neurons from CMT2B mutant mice.

E18 hippocampal neurons from + /+, fl/+, fl/fl were dissected, cultured on PLL-coated cover-glasses and maintained as described in Materials and Methods. At DIV4-7, neurons were treated with JC9 and live-imaged. a–c: Representative images of mitochondria in neurites of E18 hippocampal neurons of + /+, fl/+ and fl/fl. d: Ratiometric measurements of MMP. e: Histogram analysis of MMP distribution. In f, Representative images of mitochondrial morphology in hippocampal neurons of + /+, fl/+, fl/fl. g: Aspect ratio measurements. Mitochondrial transport in neurites of hippocampal neurons of + /+, fl/+, fl/fl was also captured by time lapsed imaging. The numbers of mitochondria in G were analyzed are: n = 88 for + /+, n = 106 for fl/+, n = 89 for fl/fl. The image series were used to generate kymographs and a representative image for each genotype is shown in h. White arrows indicate trafficking in anterograde direction, yellow arrows indicate trafficking in retrograde direction. Average velocities of axonal mitochondria moving in the anterograde (The numbers of mitochondria were analyzed are: n = 57 for + /+, n = 89 for fl/+, n = 89 for fl/fl) (i) and retrograde (The numbers of mitochondria were analyzed are: n = 68 for + /+, n = 97 for fl/+, n = 76 for fl/fl) (j) direction. k: % of retrogradely moving mitochondria. The numbers of axons in k were analyzed are: n = 22 for + /+, n = 26 for fl/+, n = 27 for fl/fl. l: % of total moving mitochondria. The numbers of axons in l were analyzed are: n = 23 for + /+, n = 26 for fl/+, n = 25 for fl/fl. Results are shown as mean ± SEM. Significance analysis was carried out using Prism. Statistical significances were calculated by One-Way ANOVA. n.s. = non significance. All p values are shown in the graphs.