| 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].,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* |
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| 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） |
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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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