文章摘要
郭亚南,王丛丛,刘云,武晓会,孙建飞,陈程,许强华.miR-210-5p对南极独角雪冰鱼心脏发育的作用机制研究[J].上海海洋大学学报,2016,25(4):481-487.
GUO Yanan,WANG Congcong,LIU Yun,WU Xiaohui,SUN Jianfei,CHEN Cheng,XU Qianghua.Functional mechanism researches of the Chionodraco hamatus miR-210-5p on the cardiac development[J].Journal of Shanghai Ocean University,2016,25(4):481-487.
miR-210-5p对南极独角雪冰鱼心脏发育的作用机制研究
Functional mechanism researches of the Chionodraco hamatus miR-210-5p on the cardiac development
投稿时间:2016-01-07  修订日期:2016-03-27
DOI:10.12024/jsou.20160101630
中文关键词: miR-210-5p  心脏发育  tbx20  南极冰鱼  bmp/smad通路
英文关键词: miR-210-5p  cardiac development  tbx20  Chionodraco hamatus  bmp/smad pathway
基金项目:国家自然科学基金面上项目(31572598);浦江人才计划(16PJ1404000);国家自然科学基金重大研究计划培育项目(91131006);上海市教育发展基金会和上海市教育委员会曙光计划(13SG51)
作者单位E-mail
郭亚南 上海海洋大学 海洋科学学院, 上海 201306  
王丛丛 上海海洋大学 海洋科学学院, 上海 201306
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 
 
刘云 上海海洋大学 海洋科学学院, 上海 201306  
武晓会 上海海洋大学 海洋科学学院, 上海 201306  
孙建飞 上海海洋大学 海洋科学学院, 上海 201306  
陈程 上海海洋大学 海洋科学学院, 上海 201306
远洋渔业协同创新中心, 上海 201306 
 
许强华 上海海洋大学 海洋科学学院, 上海 201306
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
远洋渔业协同创新中心, 上海 201306
国家远洋渔业工程技术研究中心, 上海 201306 
qhxu@shou.edu.cn 
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中文摘要:
      microRNA为短链非编码RNA,通过与靶基因3'UTR序列互补在转录后水平发挥作用。已有研究表明,microRNA在心脏发育过程中起着重要的调控作用。南极冰鱼因体内缺乏功能性血红细胞,其心脏出现了补偿性增生。前期的研究提示,独角雪冰鱼心脏中特异表达的microRNAs可能与冰鱼心脏的补偿性增生相关。本研究针对南极冰鱼心脏中高表达的miR-210-5p,运用斑马鱼显微注射、靶基因预测等手段研究了miR-210-5p对心脏发育的作用机制。结果表明:斑马鱼胚胎注射miR-210-5p后,出现心包膜水肿,心脏发育畸形等现象。qRT-PCR分析显示,过表达miR-210-5p的斑马鱼胚胎中,心脏发育相关的标志性基因bmp4smad1gata6以及tbx2b的表达水平下调。Western blotting分析发现,bmp/smad通路中的BMP2、BMP4、SMAD1、GATA6以及TBX2B的蛋白表达水平也显著下调。通过对tbx20基因3'UTR的靶基因生物信息学预测,以及对其进行GFP荧光表达分析,发现tbx20基因可能是miR-210-5p的一个靶基因。由此推测,miR-210-5p可能通过抑制tbx20基因的表达,并调控bmp/smad通路以抑制冰鱼心脏发育。
英文摘要:
      MicroRNA, a short non-coding RNA chain, plays important roles in post-transcription regulations by targeting gene 3'UTR through sequence complementary principle. Previous researches showed that microRNAs play important roles in heart development process. Heart development of the Antarctic icefish (Chionodraco hamatus) shows compensatory physiological enlargements due to the lack of functional red blood cells in the body. Previous studies suggested that microRNAs specifically expressed in the C. hamatus heart may be associated with icefish compensatory physiological enlargement. In this study, according to the highly expressed miR-210-5p, we studied the heart development functional mechanism of miR-210-5p by using zebrafish microinjection and gene's target prediction. The results showed that overexpression of miR-210-5p in 72hdf zebrafish embryos could affect zebrafish heart development, characterized by cardiac malformations such as pericardial edema. QRT-PCR analysis showed that overexpression of miR-210-5p in zebrafish downregulated heart development marker genes including bmp4, smad1, gata6, tbx2b. Western blot analysis found that the protein expression levels of BMP4, BMP2, SMAD1, GATA6 and TBX2B were downregulated. According to the bioinformatics prediction and GFP fluorescence analysis, we found tbx20 may be a target of miR-210-5p. Therefore, we speculated that miR-210-5p might inhibit tbx20 gene expression and might suppress heart development of C. hamatus through bmp/smad pathway.
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