Figure 4
From: Rewiring of regenerated axons by combining treadmill training with semaphorin3A inhibition
Histological analyses of the treatment effects on microvasculature and remyelination in the spinal cord. (A) Visualization of blood vessels using an anti-RECA-1 antibody. Images in the upper row are low-magnification views of the gray matter areas of sagittal sections immunostained for RECA-1 at 1 mm caudal to the transected site. Scale bars = 50 μm. Images in the lower row are high-magnification views that correspond to the boxed areas in the upper row images. Scale bars = 50 μm. (B) Representative image of a blood vessel with a lumen with a diameter larger than 20 μm (arrow), which indicated newly formed blood vessels following injury. Scale bars = 50 μm. Arrows in (A) also represent blood vessels with lumen diameters larger than 20 μm. The left side is rostral (A,B). (C) Quantitative analysis of RECA-1-positive areas in each group. (D) Quantitative analysis of the total areas of RECA-1-positive blood vessels with lumen diameters larger than 20 μm. *P < 0.05, **P < 0.01. Statistical analyses were based on one-way ANOVA and Bonferroni post hoc analyses. (E-M) Analyses of remyelination performed using immunohistochemistry against MBP or electron microscopy 12 weeks post-injury. (E,F,H,I,K,L) Reconstructed confocal images showing double staining (sagittal sections) for MBP (green) and GAP43 (red) in the control group (E,F), SM-345431 treatment group (H,I) and combined group (K, L). F, I and L show magnified images of the boxed areas in E, H and K, respectively. Scale bars = 100 μm. The arrow in F shows a non-myelinated (MBP-negative) GAP-43-positive axon, and the arrows in I and L show myelinated (MBP-positive) GAP-43-positive axons. The left side is rostral. (G,J) Electron microscopic images of transverse sections from the control group (G) and SM-345431 treatment group (J) at the lesion site. Scale bars = 2 μm. (N) Statistical analysis of the number of myelinated (MBP-positive) GAP-43-positive axons in each group, which were analyzed by immunohistochemistry. *P < 0.05, **P < 0.01. Statistical analyses were performed using one-way ANOVA and Bonferroni post hoc analyses. All the data are represented as the mean ± S.E.M.