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Knockdown of the neuronal nitric oxide synthase gene retard the development of the cerebellar granule neurons in vitro
Author(s) -
Li Mei,
Wang Lin,
Peng Ying,
Wang JiaChuan,
Zhou LiHua
Publication year - 2010
Publication title -
developmental dynamics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.634
H-Index - 141
eISSN - 1097-0177
pISSN - 1058-8388
DOI - 10.1002/dvdy.22189
Subject(s) - gene knockdown , biology , endogeny , nitric oxide synthase , microbiology and biotechnology , cerebellum , viability assay , in situ hybridization , sodium nitroprusside , gene expression , nitric oxide , in vitro , gene , biochemistry , neuroscience , endocrinology
The role of endogenous neuronal nitric oxide synthase (nNOS) gene in the development of cerebellar granule neurons (CGNs) is conflicting. Here, we tested the effect of antisense oligos (AS‐ODN) on the endogenous nNOS gene and the development of the CGNs in vitro. The expression of nNOS increased in a development‐dependent pattern both in terms of mRNA and protein. AS‐ODN down‐regulated nNOS gene, but in a posttranscriptional manner. Knockdown of nNOS protein decreased the viability of the CGNs from 7 to 13 days in culture (DIC). This activity of AS‐ODN was mimicked by nNOS inhibitor I. The antagonist (nNOSi, MK‐801, or ODQ) ‐induced decrease of cell viability was normalized by the provision of the sodium nitroprusside, an NO donor. This study provides direct evidence that endogenous nNOS, mainly by means of its principal product NO, plays an active role in sustaining the survival of developing CGNs at transition from differentiation to maturation. Developmental Dynamics 239:474–481, 2010. © 2009 Wiley‐Liss, Inc.