TRP-ERK通路在低温解除中国卤虫休眠胚胎滞育过程中的作用及分子机制

项目名称： TRP-ERK通路在低温解除中国卤虫休眠胚胎滞育过程中的作用及分子机制

项目编号： No.31272644

项目类型： 面上项目

立项/批准年度： 2013

项目学科： 农业科学

项目作者： 侯林

作者单位： 辽宁师范大学

项目金额： 80万元

中文摘要： 低温冷冻可以诱导中国卤虫休眠胚胎解除滞育，但终止滞育的分子机制目前还没有搞清楚。我们的前期工作发现trp、erk和gk等基因在低温诱导卤虫胚胎解除滞育过程中出现明显上调，提示其在低温诱导卤虫胚胎终止滞育中起到重要作用。本课题在我们前期得到的终止滞育相关基因trp、erk、gk、sk和sd研究的基础上，拟采用RACE技术获得各基因的全长；以Real-Time PCR法分析各基因在低温解除卤虫胚胎滞育各时期的表达模式；以原位杂交和免疫组化法分析各基因及蛋白在卤虫胚胎的表达部位；采用Western-Blot等方法确定上述各基因的相互关系及与ERK通路、糖原-山梨醇代谢、细胞周期相关通路的关系，以siRNA法研究各基因的功能，从而进一步确定由trp-erk通路调控的卤虫终止滞育的分子机制。上述研究为完善甲壳动物终止滞育的分子机制理论具有重要的理论意义，在卤虫资源的开发利用方面具有一定的应用价值。

中文关键词： 中国卤虫；trp-erk调控途径；胚胎滞育解除；早期胚胎发育；分子机制

英文摘要： Diapause termination of dormant embryo from Artemia sinica may be induced by Low temperature and freeze, but the morlecular mechanism of diapause terminated still remains unknown. In our previous researches, we found there was obvious up-regulation of expression level of trp,erk and gk genes when low temperature induced diapause terminated of dormant embryo from A. sinica, which suggested these genes play important roles in diapause termination of dormant embryo. In the project, based on the previous work described above, rapid-amplification of cDNA ends (RACE)technique will be used to obtain the full-length cDNA of each gene; Real-time PCR methord will be used to analyze the expression pattern of these genes in diapause termination of dormant embryo.The expression location in A. sinica embryo of these genes and corresponding proteins will be investigated by in situ hybridization and immunohistochemistry assay. The relationship among these genes,ERK pathway, glycol-sorbitol metabolic enzyme and cell cycle pathway will be confirmed using Western-Blot analysis. The functions of these genes will be analyzed by siRNA gene-silencing assay. The molecular mechanism of diapause termination will be demonstrated further through above experiments. All these researches have important significance of explaining the molecula

英文关键词： Artemia sinica；trp-erk regulation pathway；diapause termination；early embryonic development；molecular mechanism