Zfp36l2在胚胎体轴形成中的作用机理研究

项目名称： Zfp36l2在胚胎体轴形成中的作用机理研究

项目编号： No.31272395

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： 李红岩

作者单位： 中国海洋大学

项目金额： 82万元

中文摘要： RNA结合蛋白在早期发育中起重要调控作用，但这些转录后调控因子在脊椎动物体轴形成中的作用还不清楚。我们已克隆了斑马鱼和非洲爪蛙的Zfp36l2，发现其在中胚层特异表达，过量表达时胚胎头部发育缺陷，基因敲降时胚胎出现背部化表型，报告基因实验结果表明其可能影响经典Wnt信号通路，提示Zfp36l2可能是通过经典Wnt信号通路调节体轴形成的新基因。但其具体作用机制尚不明确。因此，我们拟首先采用RIP-差异筛选Zfp36l2的候选靶RNA；采用过量表达与基因敲降方法系统研究Zfp36l2与靶RNA的功能、与Wnt信号通路的关系以及它们在发育过程中的相互作用；采用报告基因、Western、RIP等方法确定Zfp36L2与靶RNA的结合位点及作用机制；同时筛选可能与Zfp36l2结合的蛋白。由此，阐明Zfp36l2在体轴形成中的作用机制，确定其在经典Wnt信号通路所处位置，完善体轴形成基因调控网络。

中文关键词： 斑马鱼；爪蛙；体轴形成；Zfp36l2；Wnt信号通路

英文摘要： During embryonic development, a precise and tightly regulated gene expression is necessary to ensure the proper formation of all cell types and organs. RNA binding proteins play key roles in post-transcriptional regulation of gene expression, and recent studies have shown that these proteins are involved in many developmental processes. Axis formation is an important process in embryonic development. The main signal pathway and many important transcription factors involved in axis formation have been identified, but little information is available to date regarding the functions of RNA binding proteins in vertebrate axis formation. We have identified Zfp36l2, a RNA-binding protein, in frog (Xenopus laevis) and zebrafish (Danio rerio), and whole mount in situ hybridization showed that Zfp36l2 is specifically expressed in mesoderm. We also found that Zfp36l2 mRNA overexpression resulted in head defects. In addition, luciferase experiment revealed that Zfp36l2 was able to regulate Wnt signal pathway, and knockdown of Zfp36l2 led to dorsalized embryos with enlarged head and truncated tail. Taken together, these preliminary results suggest that Zfp36l2 may play a role in axis formation, possibly by targeting RNAs involved in canonical Wnt singal pathway. However, its precise function and mode of action remain enigma

英文关键词： zebrafish；xenopus；axis formation；zfp36l2；Wnt signaling pathway