斑马鱼不同发育时期血管内皮生长因子受体2基因表达的实时定量PCR分析

Real-time Quantitative PCR Analysis of Vascular Endothelial Growth Factor Receptor-2 Expression at Different Developmental Stages of Zebrafish

[目的]探讨血管内皮生长因子受体2（VEGFR2-）基因在斑马鱼不同发育时期的表达。[方法]分别从12、24、48、72和96 h的斑马鱼胚胎和仔鱼中提取总RNA,用实时定量RT-PCR方法检测VEGFR2-基因表达,采用2^-△△Ct法进行数据分析。[结果]VEGFR2-基因表达量在12-72 h呈上升趋势,在96 h有所下降。12 h表达量最低,72 h表达量最高,与其他发育期均有显著差异。[结论]斑马鱼血管发育至72 h达成熟阶段。血管发育成熟前,VEGFR2-基因表达水平逐步增长;血管发育成熟后,VEGFR2-基因表达水平下降。

[Objective] The research aimed to investigate the expression of vascular endothelial growth factor-2（VEGF-2） in zebrafish in different developmental stages.[Method] Total RNAs were extracted from 12,24,48,72 and 96 h zebrafish embryos and larvae.Real-time quantitative RT-PCR was performed to examine the expression level of VEGFR-2.The data were analyzed by 2^-△△Ct method.[Result] The expression level of VEGFR-2 gene increased gradually from 12 h to 72 h and subsequently decreased at 96 h.The expression level was lowest at 12 h and highest at 72 h,with significant difference with that of other developmental stages.[Conclusion] The expression level of VEGFR-2 increased gradually before vessel maturation and decreased as vessels mature.