FIGURE 4.

Protection from glutamate-induced mitochondrial dysfunction and neuronal death by MFN2 in primary spinal cord motor neurons. Primary mouse spinal cord motor neurons (DIV7) isolated from Tg neurons and NTg neurons of transgenic mice overexpressing human MFN2 were treated with or without 25 μm glutamate for 24 h. A, representative immunoblot and quantification analysis confirming the overexpression of MFN2 (relative to actin) in Tg motor neurons are shown. Equal protein amounts (10 μg) were loaded and confirmed by actin. Measurement of mitochondrial length by mitoDsRed2 labeling (B) and intracellular levels of ROS (C), MMP (D), OCR (E and F), ATP (G), and neuronal death by LDH assays (H). All experiments were repeated three times. Data are means ± S.E. Statistics: one-way ANOVA followed by Tukey's multiple comparison test. *, p < 0.05; **, p < 0.01; ***, p < 0.001; ns, nonsignificant.