White matter and cerebral cortex are reduced in adult mTrappc9 deficient mice. (A-B), NFH (axonal tracts) and MBP (myelin) immunostaining of the corpus callosum sections. Compared with WT (C57BL/6 wildtype) and mTrappc9+/+, the corpus callosum (highlighted with white dotted lines) of mTrappc9m/m mice was significantly thinner. Representative images with indicated thickness from n=3 mice/group (A), and quantitative analysis of n=5 sections (B). (C-D), MBP immunostaining of the cerebral cortex sections, with the white line arrows defining the representative thickness of the cerebral cortex (C, n=3 mice). Quantification shows the cortex is significantly thinner in mTrappc9 m/m mice than the other two groups (D, n=6 sections). (E-F), MBP immunostaining of the striatum. The white dotted lines demarcate the striatum area (E, n=3 mice) for quantification of the coverage area of the nerve bundles in the striatum (F, n=6 sections), which shows that nerve bundles are significantly reduced in mTrappc9 m/m mice compared to WT and mTrappc9+/+ mice. (G-I), NFH immunostaining of spinal cord sections. In mTrappc9 m/m mice, there was no significant difference in gray matter (inside the white dotted line), but the area of white matter (outside the white dotted line) was reduced significantly. Representative images from n=3 mice/group (G); n=7 sections (H, I). (J-K), Western blot analysis of MBP and NFH protein expression in cerebral cortex normalized with GAPDH. Both showed a significant decrease in mTrappc9 m/m mice. n=3 mice/group. Data are means ± SEM; One-way ANOVA (B, D, F, H, I); Two-way ANOVA (K); ****P≤0.0001; ns, not significant (P>0.05). Scale bars: 50 μm (A); 150 μm (C, E, G).
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