CaMKII磷酸化4.1N蛋白调控GluR1转运及其分子机制研究

项目名称： CaMKII磷酸化4.1N蛋白调控GluR1转运及其分子机制研究

项目编号： No.31200807

项目类型： 青年科学基金项目

立项/批准年度： 2013

项目学科： 神经、认识与心理学

项目作者： 颜景芝

作者单位： 徐州医学院

项目金额： 26万元

中文摘要： AMPA受体GluR1亚基特异性的转运至突触后膜是LTP形成的重要机制之一，但其转运机制尚需进一步阐释。因此，研究突触可塑性的分子机制具有重要意义。以往的研究表明，GluR1的磷酸化和PDZ蛋白可以调节AMPA受体通道特性及其转运，但这些并不是AMPA受体上膜的充分条件，提示还有其它的可能机制调节AMPA受体的转运。本申请拟在本人前期研究基础上，以大鼠海马脑片和培养的海马神经元为研究对象，综合运用电生理、免疫印迹、免疫沉淀、免疫荧光、细胞表面生物素标记及活细胞工作站等方法，结合干扰小肽和RNA干扰及分子克隆的实验，试图阐明LTP过程中CaMKII激活促进4.1N蛋白Ser/Thr磷酸化，增加CaMKII-4.1N-GluR1的相互作用，从而调节GluR1至突触后膜的转运机制。本研究将进一步从分子水平揭示AMPA受体转运的分子机制。

中文关键词： 长时程增强（LTP）；GluA1；转运；4.1N；CaMKⅡ

英文摘要： The specific trafficking to postsynaptic membrane of AMPA receptor GluR1 subunit could be a major mechanism of LTP. However, the detailed mechanism of GluR1 trafficking remained to be clarified. Therefore, to study the molecular mechanisms of synaptic plasticity has an important significance. Previous studies have shown that GluR1 phosphorylation and PDZ proteins can regulate AMPA receptor channel characteristics and trafficking, but these are not a sufficient condition of the AMPA receptors on the membrane, suggesting that there are other possible mechanisms regulating the trafficking of AMPA receptor. In this application, we attempt to clarify the LTP process in CaMKII activation contributes to the 4.1N protein Ser/Thr phosphorylation, an increase of CaMKII-4.1N-GluR1-interactions, thus regulating the transport mechanism of GluR1 to the postsynaptic membrane with the integrated use of electrophysiological, western blot, immunoprecipitation, immunofluorescence, cell surface biotin-labeled and live cell station approach, combined with the interference of small peptides, RNA interference machine and molecular cloning experiments in rat hippocampal slices and cultured hippocampal neurons. This study will further reveal the molecular mechanism of AMPA receptor transport from the molecular level.

英文关键词： LTP；GluA1；trafficking；4.1N；CaMKⅡ