海七鳃鳗pma-miR-200c-3p对斑马鱼心脏发育的作用研究  

Functional effects of pma-miR-200c-3p of sea lamprey on heart development of zebrafish

作　　者：刘云[1] 王丛丛[1,2] 郭亚南[1] 许强华[1,2,3,4] 

机构地区：[1]上海海洋大学海洋科学学院,上海201306 [2]大洋渔业资源可持续开发省部共建教育部重点实验室,上海201306 [3]国家远洋渔业工程技术研究中心,上海201306 [4]远洋渔业协同创新中心,上海201306

出　　处：《大连海洋大学学报》2016年第5期510-515,共6页Journal of Dalian Ocean University

基　　金：国家自然科学基金面上项目(31572598);上海市教育发展基金会和上海市教育委员会曙光计划项目(13SG51);教育部科学技术研究项目(213013A);上海市教委水产学高峰学科项目

摘　　要：为研究海七鳃鳗Petromyzon marinus pma-miR-200c-3p在心脏发育过程中的生物学功能,采用显微注射技术,将pma-miR-200c-3p模拟体注射到斑马鱼Danio rerio胚胎,过表达pma-miR-200c-3p后,使用qRT-PCR及Western blot法检测了一些与心脏发育相关的标志性基因的表达水平,以及BMP/Smad通路重要基因蛋白水平的变化,并利用生物信息学预测、GFP荧光表达分析了pma-miR-200c-3p的生物学作用。结果表明:过表达pma-miR-200c-3p的斑马鱼胚胎中,与心脏发育相关的标志性bmp4、bmp5、smad1、gata5、tbx2b和notch1a基因的表达水平极显著上调(P<0.01),同时BMP/Smad通路中BMP4、Smad1、GATA5蛋白表达水平也明显升高;细胞试验表明,cdx1b基因可能是pma-miR-200c-3p的一个靶基因。研究表明,pma-miR-200c-3p可能通过抑制cdx1b基因的表达,参与BMP/Smad通路以促进心脏发育。The pma-miR-200c-3p mimics from sea lamprey Petromyzon marinus were injected into zebrafish Danio rerio embryos by microinjection,and qRT-PCR and Western blot were used to analyze the marker gene expression of the development of heart,and the expression levels of several genes in BMP/Smad pathway to study the biological function of pma-miR-200c-3p of the sea lamprey. Besides,target gene bio-informatics and GFP fluorescence expression analysis were used to study the function of pma-miR-200c-3p. The results showed that the gene expression levels of some cardiac development markers including bmp4,bmp5,smad1,gata5,tbx2 b and notch1 a were significantly increased in the pma-miR-200c-3p-injected embryos（ P〈 0. 01）. The protein expression levels of the BMP4,Smad1 and GATA5 were significantly increased bya BMP/Smad pathway,and it appears that cdx1 b is a target gene of pma-miR-200c-3p. Therefore,the findings indicate that pma-miR-200c-3p might repress the gene expression of cdx1 b,and be involved in the BMP/Smadpathway to promote the cardiac development process.

关 键 词：海七鳃鳗 miR-200c-3p 斑马鱼 心脏发育 cdx1b 

分 类 号：Q951[生物学—动物学]