生长激素促进斑马鱼尾鳍的再生

Growth hormone promotes regeneration of the zebrafish caudal fin

目的建立生长激素过表达的转基因斑马鱼,研究生长激素在斑马鱼尾鳍再生过程中的作用。方法利用Gateway技术构建表达质粒"pDestTol2CG2; ubi:GH-polyA",在一细胞期显微注射表达质粒和转座酶mRNA后,通过荧光显微镜和qPCR技术筛选鉴定GH过表达的转基因斑马鱼。将斑马鱼分为对照组(野生型)和生长激素过表达组,尾鳍横切后,记录分析斑马鱼尾鳍再生过程。结果转基因斑马鱼中心脏被绿色荧光蛋白标记。荧光定量PCR检测结果显示GH表达水平显著高于对照组(P<0.05)。斑马鱼尾鳍横断后,生长激素过表达组的再生速度显著提高(P<0.05)。结论建立稳定生长激素过表达的转基因斑马鱼品系,过表达生长激素能够提高斑马鱼尾鳍再生速度。

Objective To generate a transgenic zebrafish line expressing growth hormone (GH) and evaluate the effects of growth hormone (GH) overexpression on zebrafish caudal fin regeneration. Methods An expression plasmid (pDestTol2CG2;ubi:GH-polyA) was constructed by Gateway technology. After coinjection of the expression plasmid and transposase-encoding mRNA at the one cell stage, GH-overexpressing transgenic zebrafish were screened by fluorescence microscopy and qPCR. The zebrafish were divided into wild type control and GH overexpression groups. After the zebrafish caudal fin was transected, its regeneration process was recorded and analyzed. Results The heart of the transgenic zebrafish was labeled with green fluorescent protein. The real-time PCR showed that the expression level of GH was significantly higher than that in the wild type zebrafish ( P < 0.05). After the zebrafish caudal fin was transected, the regeneration rate of the GH overexpression group was increased significantly ( P < 0.05). Conclusions In this study, a transgenic zebrafish strain overexpressing GH is generated, and it demonstrates that GH overexpression promotes the regeneration of zebrafish caudal fin.