Fig. 7: CRH-dependent c-Abl activation leads to mitochondrial fission, inducing mitochondrial network fragmentation.

A IHC for MAP2 (gray) and c-Abl^T754 (red) in primary hippocampal neurons after different experimental conditions using IMATINIB (ITB), a selective c-Abl blocker (scale bar = 30 μm): vehicle, vehicle + ITB 3 μM, CRH 100 nM for 0.5 h, CRH 100 nM + ITB 3 μM and CRH 100 nM + CRHR1 Blocker NBI30775 100 nM with relative c-Abl^T754 intensity analysis. Nine different neurons acquired from three different wells were analyzed for each condition in each independent experiment. B WB analysis and quantification of DRP1^S616 expression. C Somatic mitochondrial network complexity analysis after IHC, using the same experimental conditions (scale bar = 5 μm): MAP2 (gray), mitochondria labeled with MitoTracker^TM (red), mitochondrial skeletonized structures (green) (white arrows indicates examples of networks and IS) and D–F relative network parameters quantification. Somatic mitochondrial networks were analyzed in six neurons for each condition for each different independent preparation. Experiments were performed in N = 3 independent replicates at DIV14. Data are displayed as Mean ± SEM; one-way ANOVA and Bonferroni’s post hoc comparison test were performed; data non normally distributed were analyzed by Kruskal–Wallis non parametric test followed by Uncorrected Dunn’s multiple comparison test. (*p < 0.05, **p < 0.005, ***p < 0.0005, ****p < 0.0001).