神经细胞发育过程中钙调蛋白激酶I表达的调控机制

项目名称： 神经细胞发育过程中钙调蛋白激酶I表达的调控机制

项目编号： No.31271134

项目类型： 面上项目

立项/批准年度： 2013

项目学科： 生物科学

项目作者： 王惠敏

作者单位： 华东师范大学

项目金额： 85万元

中文摘要： 本项目拟探讨在神经细胞生长发育过程中,进化上保守的非编码DNA序列以及它们的互作蛋白是如何调节钙调蛋白激酶Igamma的表达以及对神经细胞发育所起的作用。通过鉴定CaMKIgamma基因的增强子序列以及其特定的调节蛋白，了解增强子和调节蛋白因子在神经细胞发育尤其是树突发育中的功能，并阐明神经细胞发育及树突发育的分子细胞机制。在我们的前期实验中，我们发现至少两个候选CaMKIgamma增强子序列（CR1和CR5）引导的GFP表达模式与该基因表达模式相吻合。此外，我们还观察到在脑室壁和海马神经元祖细胞内以及SVZ和RMS星型胶质细胞内观均有CR1和CR5引导的GFP表达。这些前期结果表明，CR1和CR5很可能是参与神经细胞发育过程的增强子。本项目将进一步确认与神经元发育尤其是树突发育相关的增强子序列，鉴定增强子结合蛋白，并在功能上确认增强子及其结合蛋白在神经细胞发育过程中的作用和机制。

中文关键词： 钙调蛋白激酶I；树突发育；基因表达；增强子；微小核糖核酸

英文摘要： The goal of the proposed project is to determine the role of evolutionarily conserved non-coding DNA sequences and their interacting protein factors in regulating Ca2+/calmodulin-dependent protein kinaseIgamma(CaMKIgamma) expression during neuronal development. By identifying the enhancers and their trans-acting factors of CaMKIgamma, defining the function of enhancers and trans-acting factors during neuronal development, especially during dendritic development, are expected to provide novel understanding of the contribution of enhancers to neuronal development. In our preliminary experiments, we have identified two enhancer candidates (CR1 and CR5), which reproducibly drive the reporter gene (GFP) expression in mouse brain in a similar manner as CaMKIgamma?? expression.Moreover, our analysis of the GFP expression and antibody staining patter shows that GFP+ cells in VZ and hippocampus are also positively stained with DCX, whereas GFP+ cells in SVZ and RMS are positively stained with GFAP and nestin, suggesting that CR1 and CR5 maybe novel enhancers of CaMKIgamma for neuronal development. We will further define the enhancers of CaMKIgamma for dendritic development, identify enhancer-binding proteins and define the functional role of CaMKIgamma enhancers/transacting factors during neuronal development.

英文关键词： CaMKIγ；dentritic development；gene expression；enhancer；microRNA