心脏特异表达绿色荧光斑马鱼模型的建立与评估  

Construction and assessment of heart-specific green fluorescence zebrafish line

作　　者：彭夕洋[1] 陈婷芳[1] 黄婷[1] 江志钢[1] 吴秀山[1] 邓云[1] 

机构地区：[1]湖南师范大学心脏发育研究中心,发育生物学与蛋白质组学教育部重点实验室,长沙410081

出　　处：《遗传》2013年第4期511-518,共8页Hereditas（Beijing)

基　　金：国家自然科学基金项目(编号:81170088;30970245;3093005;31171402)资助

摘　　要：本课题组前期研究中,利用斑马鱼cmlc2(Cardiac myosin light chain 2)基因启动子构建了一个用于斑马鱼心脏组织特异表达外源基因的转基因表达载体pTol2-cmlc2-IRES-EGFP。文章利用该载体构建了一个稳定表达EGFP的转基因斑马鱼品系,并初步分析了EGFP的表达对该转基因斑马鱼品系的心脏发育和功能的影响。结果表明,在建立的转基因斑马鱼品系早期胚胎发育过程中,绿色荧光信号在心脏中特异表达,该表达模式与原位杂交分析的cmlc2的表达模式结果相同;该转基因斑马鱼品系的心脏形态及发育生长正常;进一步通过M-Mode分析心脏生理学功能的结果表明:该转基因品系心动周期、心率、收缩与舒张表面积及表面积缩短率等重要生理指标与正常野生型的斑马鱼对照组相比没有显著差别。以上结果表明该转基因品系中绿色荧光蛋白的表达对斑马鱼心脏的发育和功能没有影响。研究结果为进一步利用该载体建立外源目的基因转基因表达模型,研究心脏表达基因的功能奠定了重要基础。Using the promoter for cardiac myosin light chain 2 （cmlc2） gene, an expression vector pTol2-cmlc2-IRES- EGFP for making heart-specific expression of exogenous gene in transgenic zebrafish was generated previously. Here, we reported the construction of a transgenic zebrafish line which stably expresses EGFP using this vector, and the effects of EGFP on the heart development and cardiac function of this transgenic zebrafish line were preliminarily analyzed. The re- sults showed that the green fluorescence signal of cmlc2：EGFP line under fluorescence microscopy specifically expressed in heart and faithfully recapitulated both the spatial and temporal expression pattems of endogenous cmlc2 gene revealed by in situ hybridization in the early developmental stages. The cardiac morphology and development of this transgenic zebrafishline remained to be normal. Furthermore, the heart morphology and physiological function of this transgenic line have been analyzed using M-mode analysis. The results showed that there was no significant difference between the cmlc2：EGFP and the wild type lines with respect to heart period, heart rate, diastolic surface area and systolic surface area, and fractional area change. No tachyarrhythmia was observed in the embryos from either line. Thus, the excessive expression of EGFP in this transgenic line seemed to exert no detrimental effects on the function and development of zebrfish hearts during early stages Our study laid a foundation for the construction of exogenous gene transgenic line using pTol2-cmlc2-IRES-EGFP vector to study the function of genes that expressed in heart.

关 键 词：斑马鱼 转基因 EGFP Tol2转座子 

分 类 号：R-332[医药卫生]