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海马神经元中糖皮质激素受体表达的培养条件和胚胎阶段依赖性沉默

Culture condition and embryonic stage dependent silence of glucocorticoid receptor expression in hippocampal neurons.

作者信息

Xiao Lin, Chen Yizhang

机构信息

Institute of Neuroscience, Department of Neurobiology, Second Military Medical University, 800 XiangYin Road, Shanghai 200433, PR China.

出版信息

J Steroid Biochem Mol Biol. 2008 Jul;111(1-2):147-55. doi: 10.1016/j.jsbmb.2008.03.036. Epub 2008 Jun 25.

Abstract

Glucocorticoid (GC) plays a key role in controlling numerous cellular processes during embryogenesis and fetal development. The actions of glucocorticoids are mediated by interaction with their receptors. We previously reported that hippocampal neurons from embryonic day 18 (E18) rats showed silence of glucocorticoid receptor (GR) expression when cultured in serum-free condition. In this study, using western blot, immunofluorescence staining and real-time RT-PCR, we found that while this silence occurred in hippocampal neurons isolated from E16 and E18 rats, it did not happen in those from E20 and neonatal (P0) rats. And when cultured under serum-containing condition, none of them showed GR silence anymore. Corticosterone could not rescue the expression of GR in E16 and E18 neurons in serum-free condition, whereas adding of serum could induce the re-expression of the silenced GR. The absence of GR silence in P0 neurons was not due to the perturbation during parturition. Moreover, the unique expression profile of GR in protein and mRNA level was well reflected in the changes of GR function. These results suggested that under in vitro condition, serum was critical for the maintaining of GR expression in hippocampal neurons of early embryonic stages but less important in later developmental stages. Thus, our data implied that at different developmental stages, the expression of GR in hippocampal neurons might have different susceptibilities to environment changes and there might be a critical time window for the switching of such characteristics during development.

摘要

糖皮质激素(GC)在胚胎发生和胎儿发育过程中控制众多细胞过程中起关键作用。糖皮质激素的作用是通过与其受体相互作用介导的。我们之前报道,在无血清条件下培养时,胚胎第18天(E18)大鼠的海马神经元显示糖皮质激素受体(GR)表达沉默。在本研究中,我们使用蛋白质免疫印迹、免疫荧光染色和实时逆转录-聚合酶链反应发现,虽然这种沉默发生在从E16和E18大鼠分离的海马神经元中,但在从E20和新生(P0)大鼠分离的海马神经元中并未发生。并且在含血清条件下培养时,它们都不再显示GR沉默。皮质酮在无血清条件下不能挽救E16和E18神经元中GR的表达,而添加血清可诱导沉默的GR重新表达。P0神经元中不存在GR沉默并非由于分娩过程中的干扰。此外,GR在蛋白质和mRNA水平上独特的表达谱在GR功能变化中得到了很好的体现。这些结果表明,在体外条件下,血清对于维持早期胚胎阶段海马神经元中GR的表达至关重要,但在后期发育阶段则不太重要。因此,我们的数据表明,在不同发育阶段,海马神经元中GR的表达可能对环境变化有不同的敏感性,并且在发育过程中可能存在一个关键的时间窗口用于这种特性的转换。

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