Ults are constant using the thought that GFs not potential of cell divisions, it remained undetermined to what extent only stimulated proliferation of endogenous NPCs, but additionally proGFP /GFAP cells reflected de novo differentiation of NPCs into moted their neuronal differentiation in vivo. GFs may possibly have supthe astrocyte lineage. ported the survival of newly generated neurons also, but such a Enhanced neurogenesis by Neurogenin2 and BDNF in vitro survival effect couldn’t completely account for the observed improve within the above study demonstrated that the production of new neuthe number of new neurons among DAI3 and DAI7. We found, rons from endogenous NPCs may be induced beneath particular conhowever, that the numbers of GFP /TuJ1 and GFP /HuC/D cells steadily decreased right after DAI7, and they at some point disapditions. This, in turn, suggests the presence of specific mechapeared by DAI28 (data not shown). Furthermore, as described nisms that actively suppress the neurogenic prospective of NPCs in above, no GFP cells were found to express NeuN, which feasitu. We initial addressed this problem employing in vitro culture of NPCs. tures a more mature phenotype of neurons, at any time points To mimic the situation of virus-infected NPCs in vivo, growing examined when manage viruses were made use of for infection (see neurosphere cells were infected with pMXIG viruses, and subsebelow). quently, neuronal and glial differentiation of GFP cells soon after removal of GFs was examined (Fig. five). Unlike these neuronal cells, substantial fractions of GFP cells expressed glial cell markers GFAP (Fig. 4G) and GalC (Fig. It has been shown that the expression of numerous cytokines is four H) with no remedy with GFs, and their percentages have been not drastically upregulated inside the injured spinal cord (Nakamura substantially distinct among Beta-2 Adrenergic Receptor Proteins Recombinant Proteins GF-treated and untreated animals and Bregman, 2001; Setoguchi et al., 2001, 2004; Velardo et al., ( p 0.160 for GFAP cells and p 0.327 for GalC cells) (Fig. 2004; Chen et al., 2005). Among them, BMPs and CNTF have 4 I). Couple of GFP or BrdU cells have been GalC at earlier time points, been shown to inhibit neuronal differentiation of NPCs each in suggesting that GFP /GalC cells detected at DAI7 have been newly vivo and in vitro (Lim et al., 2000; Nakashima et al., 2001; Setogugenerated oligodendrocytes. The truth is, it has been demonstrated chi et al., 2004). Consistent with this, therapy of neurospheres that immature oligodendrocytes are generated in both the intact with BMP4 and CNTF drastically improved the percentage ofOhori et al. Regeneration with the Injured Spinal CordJ. Neurosci., November 15, 2006 26(46):11948 1960 Figure 6. Induction of new neurons by GFs, Ngn2, and BDNF in vivo. A, Effects of GFs and Ngn2 on neuronal differentiation of GFP-labeled cells in vivo. Manage and Ngn2 viruses have been administered with (red bars) or Ubiquitin-Specific Protease 12 Proteins medchemexpress without the need of (white bars) GFs into injured spinal cords, and subsequently the percentages of HuC/D (left) and NeuN (correct) cells amongst total GFP cells have been quantified at DAI7. GFP /HuC/D cells had been detected in dissociated single cells, whereas GFP /NeuN cells were detected in tissue sections. p 0.01 compared with handle virus-infected animals. p 0.01 compared with Ngn2 with out GFs. B , Micrographs showing GFP cells (green) costained for Ngn2 (red) and NeuN (blue) (B) and BrdU (red) and MAP2 (blue) (C, C’) at DAI7, synaptophysin (red) and MAP2 (blue) (D), GABA (red) (E), and NeuN (red) (F) at DAI28. C’ shows a magnified view of a neurons indi.