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[目的]探讨血管内皮生长因子受体2(VEGFR-2)基因在斑马鱼不同发育时期的表达。[方法]采用Trizol法分别提取12、24、48、72和96h斑马鱼胚胎和仔鱼总RNA;再使用MMLV反转录酶将其反转录为cDNA;合成的cDNA用实时定量RT-PCR方法检测VEGFR-2基因的表达。实时定量RT-PCR扩增反应结束后,利用熔解曲线对扩增产物进行特异性分析。采用2-△△Ct法进行数据分析。[结果]VEGFR-2基因表达量在12～72h呈上升趋势,在96h时有所下降。12h相对表达量最低,与其他发育期差异极显著(P<0.01),72h相对表达量最高,与12、24和96h差异极显著(P<0.01),与48h差异显著(P<0.05)。[结论]斑马鱼血管发育至72h达成熟阶段,血管发育成熟前,VEGFR-2基因表达水平逐步增长;血管发育成熟后,VEGFR-2基因表达水平下降。
[Objective] To investigate the expression of vascular endothelial growth factor receptor 2 (VEGFR-2) gene in different stages of zebrafish development. [Method] The total RNA of zebrafish embryos and larvae of 12, 24, 48, 72 and 96 h were extracted by Trizol method respectively, and then reverse transcribed into cDNA using MMLV reverse transcriptase. The synthesized cDNA was analyzed by real-time quantitative RT-PCR The expression of VEGFR-2 gene was detected. After the real-time quantitative RT-PCR amplification reaction was completed, the amplification product was analyzed by melting curve. Data analysis was performed using the 2- △△ Ct method. [Result] The expression of VEGFR-2 gene increased from 12 to 72 hours and decreased at 96 hours. (P <0.01). The highest relative expression at 12h, 12h, 24h and 96h was significant (P <0.01), and the difference was significant at 48h (P <0.05). [Conclusion] The development of blood vessel of zebrafish reached the mature stage at 72h. The expression of VEGFR-2 gene gradually increased before the development of blood vessels. After the development of blood vessels, the expression of VEGFR-2 gene decreased.