Pcdh18基因抑制对小鼠胚胎中枢神经系统发育的影响

项目名称： Pcdh18基因抑制对小鼠胚胎中枢神经系统发育的影响

项目编号： No.31201864

项目类型： 青年科学基金项目

立项/批准年度： 2013

项目学科： 畜牧学与草地科学、兽医学、水产学

项目作者： 殷俊

作者单位： 扬州大学

项目金额： 23万元

中文摘要： Pcdh18是一种钙黏附蛋白。Pcdh18基因抑制可致斑马鱼神经系统发育异常并呈剂量依赖性胚胎死亡，该结果可为人类Pcdh18基因功能确定提供线索。但是，斑马鱼与人类Pcdh18仅有66%的序列同源性，且结构也不尽相同，若要获得更为可靠的参考，须以哺乳动物为对象做进一步研究。研究显示，Pcdh18极有可能是必须基因，基因敲除可因神经系统发育缺陷导致胚胎死亡，因此，获得Pcdh18基因敲除小鼠的前景不容乐观。将RNA干扰理论与转基因小鼠技术相结合，通过控制基因抑制效率获得Pcdh18恰当"下调"而非完全"敲除"的RNAi小鼠，既可规避胚胎死亡，又可获得缺陷表型进而倒推基因功能。以所建立的RNAi小鼠为模型，通过原位杂交技术追踪神经发育过程中结构标志基因的表达变化，并结合形态学观察和凋亡检测的手段，研究Pcdh18基因抑制对小鼠胚胎中枢神经系统发育的影响，其结果可为该基因功能确定提供可靠参考。

中文关键词： 原钙黏附蛋白18（pcdh18)；RNA干扰；基因功能；转基因技术；

英文摘要： Studies had shown that down regulation of protocadherin18 (Pcdh18), a kind of cadherin, induced abnormal embryonic neurogenesis and caused fetal death in a dose-dependent fashion in zebrafish, which provided clues to determine the gene function of human Pcdh18. The homology of Pcdh18 nucleotide sequence in zebrafish and human is only 66%, and their protein structures are not exactly the same. These differences made it necessary to study Pcdh18 in mammalian models to get more reliable information for determining the gene function. As the data suggested, Pcdh18 was essential for embryonic survival and loss of Pcdh18 resulted in fetal death mainly because of neurogenesis defects. Thus, it was not optimistic to obtain the Pcdh18 knockout mice by traditional knockout technology for further research. Combination of RNA interference (RNAi) theory and transgenic mice technology, a sound method was to construct Pcdh18 RNAi mice in which Pcdh18 was properly knockdown but not knockout by controlling the rate of gene inhibition. The advantage of the RNAi mice model was twofold: (1) to avoid fetal death; and (2) to produce defective phenotype for researching the gene function. Generation of Pcdh18 RNAi mice as a model, the effects of Pcdh18 down regulation on the embryonic central nervous system would be identified by morph

英文关键词： Protocadherin18 (Pcdh18)；RNA interference；Gene function；Transgenic technique；