新基因Chin1在神经细胞凋亡中的作用及其分子机制

项目名称： 新基因Chin1在神经细胞凋亡中的作用及其分子机制

项目编号： No.30800311

项目类型： 青年科学基金项目

立项/批准年度： 2009

项目学科： 生物科学

项目作者： 陈米娜

作者单位： 四川大学

项目金额： 21万元

中文摘要： 早期研究发现Chin1基因功能可能与神经系统发育相关，可能参与维持神经细胞存活和抑制神经细胞凋亡。本研究主要通过构建Chin1条件性基因敲除小鼠，利用分子、细胞生物学等方法研究Chin1基因在脑发育以及神经细胞存活、凋亡中的作用和分子机制。我们已成功构建了Chin1中枢神经系统条件性基因敲除小鼠，发现该品系小鼠脑结构并无明显异常，并且未检测到神经细胞的凋亡增加。通过GST-pulldown以及Co-IP的方法，我们对凋亡抑制蛋白Prosaposin（早期经酵母双杂交发现的Chin1可能的相互作用蛋白）与Chin1的相互作用进行验证，结果二者并无直接相互作用。通过mRNA微阵列分析Chin1中枢神经系统条件性敲除小鼠与对照小鼠的差异表达基因发现，多个髓鞘相关基因如mbp/cnp/plp/mobp等mRNA水平在敲除小鼠中下降。利用荧光定量PCR的方法对该结果得到了验证。同时观察到在少突胶质细胞分化过程中，Chin1蛋白逐渐向细胞核内转移,提示Chin1基因可能在少突胶质细胞分化方面具有重要的调节功能。根据以上实验结果,我们将重点研究Chin1基因在少突胶质细胞分化方面的功能。

中文关键词： Chin1；细胞凋亡；基因敲除；少突胶质细胞；细胞分化

英文摘要： In the early studies we found that Chin1 gene may play a role in the development of nervous system. Also, there were some data that suggested that Chin1 might involve in the maintenance of survival and the inhibition of apoptosis of the neuronal cells. Therefore, we proposed to develop Chin1 knockout mice to study Chin1 gene functions and mechanisms in the brain development and the neuronal cell survival and apoptosis. We have successfully developed central nervous system specific Chin1 knockout mice. We found that loss of Chin1 does not cause obvious abnormality of the brain nor increases neuronal cell apoptosis. To validate the interaction of the Chin1 protein and the pro-apoptotic protein Prosaposin, a potential interact protein of Chin1 indicated by Yeast two- hybrid, we performed co-IP and GST-pulldown experiments. The result showed that there is no direct interaction of the two proteins. However, we did notice that the mRNA level of multiple myelin genes such as mbp,cnp,plp and mobp etc. are down-regulated in Chin1 knockout brains by Microarray. We validated this result by real-time PCR. Furthermore, we found that the differentiation of oligodendrocyte is accompanied by the translocation of Chin1 protein from cytoplasma to the nucleus. These results indicate that Chin1 may play an important role in the regulation of oligodendrocyte differentiation. Therefore, we are going to study the functions and mechanisms of Chin1 gene in the oligodendrocyte differentiation in future.

英文关键词： Chin1;apoptosis;gene knockout;oligodendrocyte;cell differentiation