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[Preprint]. 2024 Apr 3:2024.04.02.587832. [Version 1] doi: 10.1101/2024.04.02.587832

Figure 2. Vglut2-Cre mediated OPTNΔC causes RGC and spinal cord motor neuron degeneration and ALS-like locomotor deficits.

Figure 2.

A, Left, representative in vivo OCT images of retinas from 4-week-old (4w) to 12w OPTNf/f naïve mice and OPTNf/f::Vglut2-Cre mice. Right, quantification of GCC thickness in OPTNf/f naïve and OPTNf/f::Vglut2-Cre mouse eyes. n = 2–7 mice. B, Visual acuity measured by OKR in 4w and 12w OPTNf/f naïve and OPTNf/f::Vglut2-Cre mouse eyes. n = 3–8 mice. Data are presented as means ± s.e.m, *: p<0.05, ***: p<0.001, ****: p<0.0001, Two-way ANOVA with Sidak’s multiple comparisons test. C, Left, representative confocal images of retina wholemounts showing surviving RBPMS-positive (red) RGCs at 4w and 12w. Scale bars, 20 μm; Right, quantification of surviving RGC somata, represented as percentage of OPTNf/f::Vglut2-Cre eyes compared with OPTNf/f eyes. D, Left, light microscopic images of semi-thin transverse sections of ONs with PPD staining at 4w and 12w. Scale bars, 10 μm. Right, quantification of surviving axons in ONs at 4w and 12w, represented as percentage of OPTNf/f::Vglut2-Cre eyes compared with OPTNf/f eyes. n = 8–15 mice. E, Behavioral tests of locomotion, including four-paw grip strength, distance traveled in open field test, and latency to fall in rotarod test, were performed in 4- (male = 4, female =5) and 12-weeks old (male = 6, female=1) OPTNf/f:: Vglut2-Cre mice and compared to same-age naïve OPTNf/f mice (4 weeks male = 6, female = 4; 12 weeks male = 7 or 6, female = 5). F, Immunofluorescent labeling of neurons with NeuN (green) in lumbar segments 1–3 spinal cord sections of the OPTNf/f ::Vglut2-Cre mice and same-age naïve OPTNf/f mice. The motor neurons in the ventral horns were quantified as NeuN+ and larger than 500 μm2. Scale bars, 100 μm. Quantification of motor neuron survival at 4- or 12-weeks-old are shown to the right. n = 5–12 mice. All the quantification data are presented as means ± s.e.m, *: p<0.05, **: p<0.01, ****: p<0.0001, ns: no significance, C-F with unpaired t-test.