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Tissue-Specific Stem Cells
Article first published online: 18 NOV 2010
Copyright © 2010 AlphaMed Press
Volume 28, Issue 11, pages 2041–2052, November 2010
How to Cite
Luo, C.-X., Jin, X., Cao, C.-C., Zhu, M.-M., Wang, B., Chang, L., Zhou, Q.-G., Wu, H.-Y. and Zhu, D.-Y. (2010), BIdirectional Regulation of Neurogenesis by Neuronal Nitric Oxide Synthase Derived from Neurons and Neural Stem Cells. STEM CELLS, 28: 2041–2052. doi: 10.1002/stem.522
Author contributions: C.-X.L.: conception and design, data analysis and interpretation, manuscript writing; X.J.: cell culture, cell proliferation assay, collection and assembly of data; C.-C.C.: immunofluorescence, collection and assembly of data.; M.-M.Z.: NO assay in living cells, lentivirus production and infection of cultures; B.W.: RT-PCR, telomerase activity assay; L.C.: Western blot analysis; Q.-G.Z.: Western blot analysis; H.-Y.W.: technical support; D.-Y.Z.: conception and design, data analysis and interpretation, manuscript writing.
First published online in STEM CELLS EXPRESS September 15, 2010.
Disclosure of potential conflicts of interest is found at the end of this article.
- Issue published online: 18 NOV 2010
- Article first published online: 18 NOV 2010
- Accepted manuscript online: 15 SEP 2010 01:49PM EST
- Manuscript Accepted: 2 SEP 2010
- Manuscript Received: 6 MAY 2010
- National Natural Science Foundation of China. Grant Numbers: 3097102, 30901550
- National Basic Research Program of China (973 Program. Grant Number: 2011CB504404
- Natural Science Foundation of Jiangsu Province. Grant Number: 09KJB310004
- Neuronal nitric oxide synthase;
It has been demonstrated that neuronal nitric oxide synthase (nNOS) negatively regulates adult neurogenesis. However, the cellular and molecular mechanisms underlying are poorly understood. Here, we show that nNOS from neural stem cells (NSCs) and from neurons play opposite role in regulating neurogenesis. The NSCs treated with nNOS inhibitor N5-(1-imino-3-butenyl)-L- ornithine (L-VNIO) or nNOS gene deletion exhibited significantly decreased proliferation and neuronal differentiation, indicating that NSCs-derived nNOS is essential for neurogenesis. The NSCs cocultured with neurons displayed a significantly decreased proliferation, and deleting nNOS gene in neurons or scavenging extracellular nitric oxide (NO) abolished the effects of coculture, suggesting that neurons-derived nNOS, a source of exogenous NO for NSCs, exerts a negative control on neurogenesis. Indeed, the NSCs exposed to NO donor DETA/NONOate displayed decreased proliferation and neuronal differentiation. The bidirectional regulation of neurogenesis by NSCs- and neurons-derived nNOS is probably related to their distinct subcellular localizations, mainly in nuclei for NSCs and in cytoplasm for neurons. Both L-VNIO and DETA/NONOate inhibited telomerase activity and proliferation in wild-type (WT) but not in nNOS−/− NSCs, suggesting a nNOS-telomerase signaling in neurogenesis. The NSCs exposed to DETA/NONOate exhibited reduced cAMP response element binding protein (CREB) phosphorylation, nNOS expression, and proliferation. The effects of DETA/NONOate were reversed by forskolin, an activator of CREB signaling. Moreover, disrupting CREB phosphorylation by H-89 or LV-CREB133-GFP simulated the effects of DETA/NONOate, and inhibited telomerase activity. Thus, we conclude that NSCs-derived nNOS stimulates neurogenesis via activating telomerase, whereas neurons-derived nNOS represses neurogenesis by supplying exogenous NO that hinders CREB activation, in turn, reduces nNOS expression in NSCs. STEM CELLS 2010;28:2041–2052