The expression patterns of altered miRNA target genes in injured spinal cord. A–C1: Representative microphotographs of the glial cells on the 14th day after ventral combined with dorsal root avulsion. A1, B1, C1 shows the enlarged photographs of A, B, C, respectively. The increased expression of iNOS protein was revealed in injured spinal segments (A) and was widely distributed both in the gray and white matter (A1). More obviously, GFAP immunoreactivity was observed in the gray matter of the ipsilateral ventral horns; (B) and was activated with larger cell bodies and short processes (B1). The ventral combined with dorsal root avulsion-induced nestin immunoreactivity was concentrated in the Rexed IX layer of the gray matter; (C) and showed a spider shape, larger cell bodies, and coarse and short processes (C1). D–I: Representative microphotographs of the Akt/PKB signaling pathway on the 14th day after ventral combined with dorsal root avulsion. Both Akt (D, E) and PI3K (F, G) were highly expressed in the glial cells in the bilateral ventral horns. The caspase-3 immunoreactivity was remarkably induced within the cytoplasm of the ipsilateral ventral horn motoneurons (H, I) J–O: Representative microphotographs of c-jun, ATF-3 and CREB. c-jun was highly expressed in the nuclei of the ipsilateral ventral horn motoneurons (J, K). CREB protein was distributed in the nuclei of both motoneurons and glial cells in the bilateral ventral horns (N, O). However, the expression of ATF-3 was only observed inside the motoneurons of the ipsilateral ventral horns (L, M). P–U: Representative microphotographs of calpain 2, nNOS and ChAT. Ventral combined with dorsal root avulsion resulted in a clear decrease in the expression of calpain-2 in the ipsilateral ventral horn (P, Q). The induction of nNOS was observed only in the ipsilateral ventral horn motoneurons (R, S). Ventral combined with dorsal root avulsion resulted in the disappearance of ChAT in the ipsilateral motoneurons (T, U). Scale bar = 100 μm.