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. 2012 Sep 24;287(46):38531–38542. doi: 10.1074/jbc.M111.336925

FIGURE 8.

FIGURE 8.

Intramuscular administration of ChABC is effective for skeletal muscle regeneration and improvement of mdx pathology in mice. A, transverse TA muscle sections from vehicle- and ChABC-treated wild-type mice were immunostained with anti-laminin (green) plus anti-CS stub (ΔCS, red) antibodies. Cell nuclei were stained with Hoechst 33342 and pseudo-colored red. The time schedule of injection and immunohistochemistry (IHC) is shown at the bottom of each panel. ChABC treatment degrades CS chains and produces characteristic CS-stub structures (ΔCS) around individual myofibers. Centrally nucleated myofibers were not observed in either ChABC-treated muscles or in the control. Scale bar, 100 μm. B and C, ChABC treatment promoted regeneration of wild-type TA muscles injured by CTX. B, time schedules of CTX injection with/without ChABC injection and immunohistochemistry (bottom) and representative images of lesion areas of CTX-treated TA muscles from wild-type mice. Most myofibers labeled with anti-laminin antibody (green) had central nuclei (red). Scale bar, 100 μm. C, CSA of all of the centrally nucleated myofibers within injured regions of the CTX-treated TA muscles was measured (CTX, n = 4 (the number of TA muscles analyzed), 2,381 fibers (total number of fibers analyzed); CTX ≫ ChABC, n = 4, 2,545 fibers; CTX + ChABC, n = 5, 2,552 fibers). Note a significant increment in the CSA in both ChABC-treated muscles compared with the control group treated with CTX alone (results are expressed as means ± S.D.; *, p < 0.01, CTX versus CTX ≫ ChABC or CTX + ChABC). D and E, ChABC treatment of TA muscles of mdx mice. D, transverse TA muscle sections from vehicle- and ChABC-treated mdx mice were labeled with anti-laminin antibody (green) and Hoechst 33342 (pseudocolor, red). The time schedule of injection and IHC is shown at the bottom of each panel. Scale bar, 100 μm. E, fraction of centrally nucleated myofibers (% of total fibers) within each TA muscle section obtained from mdx mice on day 7 post-treatment was measured (n = 5, for each group; results are expressed as means ± S.D.; *, p < 0.005).