Fig. 6: DRP1 mediates cell death after ONC.
From: DLK-dependent axonal mitochondrial fission drives degeneration after axotomy
a Schematic representation of RGC survival following optic nerve crush (ONC). b Western blots of Control and DRP1 gRNA transduced optic nerved. Immunoblot for DRP1 and loading control ß actin. c Quantification of DRP1 knockdown by Western blot. N = 4 samples. d Representative images of cleaved caspase 3 positive cells in saCas9-Control and saCas9-DRP1 gRNA transduced RGCs 3 days after ONC. Scalebar 25 µm. e Average cleaved caspase 3 positive cells per section in saCas9-Control, saCas9-DRP1 gRNA and EGFP transduced RGCs after nerve crush. N = 6 Control gRNA, n = 4 DRP1 gRNA and n = 6 EGFP animals per condition. One-way ANOVA, Bonferroni correction. (Control gRNA vs EGFP, not significant, ns. Control gRNA vs DRP1 gRNA p = 0.025 *. DRP1 gRNA vs EGFP p = 0.0093 **). f Representative images of retinas 7 days post ONC and immunostained for RGC marker RBPMS (magenta). Images acquired at similar areas and the same distance from the optic nerve head (ONH). Scalebar 100 µm. g Quantification of percentage RBPMS-positive RGCs in the retinas of control and DRP1 gRNA mice at 7DPI (normalized to contralateral condition). (N = 8 mice per condition. Unpaired, two-tailed, t test. p < 0.0001 ****). h Representative isodensity maps display the topographical survival of RBPMS+RGCs at 7 days post-injury. DRP1 gRNA treatment delays ONC-induced RGC degeneration across the retina. Color scale for isodensity maps ranges from 0 (purple) to 3600 (red) RGCs/mm². All graphical representations show mean ± SEM.
