Article Summary
李国然 宋晓伟 马亮亮 郭俊 马丽萍△ 赵仙先△.miR-22心肌特异转基因斑马鱼的构建及心肌肥厚的评估[J].现代生物医学进展英文版,2016,16(14):2608-2611.
miR-22心肌特异转基因斑马鱼的构建及心肌肥厚的评估
Construction of Cardiac-specific miR-22 Transgenic Zebrafish Line andEvaluation of Cardiac Hypertrophy*
  
DOI:
中文关键词: miR-22  心肌特异  转基因斑马鱼  心肌肥厚
英文关键词: miR-22  Cardiac-specific  Transgenic zebrafish line  Cardiac hyoertrophy
基金项目:国家自然科学基金面上项目(81170223)
Author NameAffiliation
李国然 宋晓伟 马亮亮 郭俊 马丽萍△ 赵仙先△ 第二军医大学长海医院心血管内科 
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中文摘要:
      目的:构建miR-22心肌特异转基因斑马鱼系,在体评估miR-22 对于心肌肥厚的作用。方法:构建pTol2-CMLC2-miR-22- IRES-EGFP表达载体。通过显微注射的方法将tol2重组质粒于一细胞期注射入斑马鱼受精卵胚胎中,荧光筛选获得心肌特异表 达绿色荧光的斑马鱼胚胎,并稳定表达传代。然后对稳定传代的成年斑马鱼心脏进行心肌肥厚及心功能的检测。结果:成功建立 了miR-22 心肌特异转基因斑马鱼系,通过定量PCR确定心肌中miR-22表达升高,荧光显微镜观察发现斑马鱼心肌出现绿色荧 光。miR-22 心脏特异过表达的转基因鱼系的成年鱼与野生对照组相比,出现了心肌肥厚的现象,心肌肥厚分子标志物nppa、myh7 明显升高。斑马鱼心脏病理切片结果同样显示出miR-22 心肌特异转基因斑马鱼出现了心肌肥厚的现象。结论:成功构建了 miR-22 心肌特异转基因斑马鱼,为研究心肌中miR-22 的生物学功能提供了重要的工具,并证明miR-22 心脏特异过表达会引起 斑马鱼心肌肥厚。
英文摘要:
      Objective:Construction of cardiac-specific miR-22 transgenic zebrafish line and evaluation of cardiac hypertrophy.Methods:To construct the pTol2-CMLC2-miR-22-IRES-EGFP expression vector, The Tol2 recombinant plasmid was injected into zebrafish embryos in one-cell stage by microinjection, and the embryos expressing cardiac-specific green fluorescence were screened. Different generation were selected and raised to get a stable transgenic line. And then heart section and analysis of heart function were performed on the stably miR-22 overexpressed adult zebrafish.Results:The cardiac-specific miR-22 transgenic zebrafish line was constructed successfully, in which over-expression of miR-22 in cardiac tissues was identified via quantitative PCR and displayed the phenomenon of cardiac hypertrophy. Fluorescence microscopy revealed a green fluorescent zebrafish heart. Molecular markers of cardiac hypertrophy, nppa and myh7, were significantly increased. miR-22 cardiac-specific transgenic zebrafish cardiac pathology results also appeared hypertrophy phenomenon.Conclusion:We successfully constructed a cardiac-specific miR-22 transgenic zebrafish to study the function of miR-22 in myocardial tissues and identified miR-22 cardiac-specific overexpression would cause cardiac hypertrophy.
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