Ex vivo and in vivo efficacy of CAH derivatives against PND. (A) Sciatic nerve explant culture was used for evaluating ex vivo efficacy. Transverse stripes of sciatic nerve explants in the presence or absence of compounds of 100 µM were observed and photographed under a stereomicroscope for three days in vitro (3DIV). Scale bar, 50 µm. The compounds were 3a, 3b, 3c, 3e, 3f, and 3g. (B) The number of transverse stripes in a 2 mm-long sciatic nerve was defined as stripe index and measured by counting under a stereomicroscope (n = 3 mice). (C) Ovoid-shaped myelin formation appeared in the sciatic nerve fibers cultured for 3DIV with or without treatment of the compounds. (D) The ovoid index was obtained by counting the ovoid-shaped myelin in 200 µm-long sciatic nerve fibers under a differential interference contrast (DIC) filtered microscope. Scale bar, 50 µm. (E) In vivo PND model using [Tg(mbp:eGFP)] zebrafish are visualized. (F) MBP: eGFP fluorescence in the caudal fin (green) with 3f treatment of 1 µM for 10 days was observed under a stereomicroscope. Scale bar, 0.25 mm. (G) The number of intact GFP fluorescence was counted in three areas (1 × 1 mm2) of the distal part of the injury. * p < 0.05, ** p < 0.01, and *** p < 0.001.
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