CAJ | 학술논문

[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96 h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12～72 h呈上升趋势,在96 h时有所下降.12 h相对表达量最低,与其他发育期差异极显著(P <0.01),72 h相对表达量最高,与12、24和96 h差异极显著( P <0.01),与48 h差异显著(P <0.05).[结论] 斑马鱼血管发育至72 h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.
[목적]탐토혈관내피생장인자수체2(VEGFR-2)기인재반마어불동발육시기적표체.[방법]채용Trizol법분별제취12、24、48、72화96 h반마어배태화자어총RNA;재사용MMLV반전록매장기반전록위cDNA;합성적cDNA용실시정량RT-PCR방법검측VEGFR-2기인적표체.실시정량RT-PCR확증반응결속후,이용용해곡선대확증산물진행특이성분석.채용2-△△Ct법진행수거분석.[결과] VEGFR-2기인표체량재12～72 h정상승추세,재96 h시유소하강.12 h상대표체량최저,여기타발육기차이겁현저(P <0.01),72 h상대표체량최고,여12、24화96 h차이겁현저( P <0.01),여48 h차이현저(P <0.05).[결론] 반마어혈관발육지72 h체성숙계단,혈관발육성숙전,VEGFR-2기인표체수평축보증장;혈관발육성숙후,VEGFR-2기인표체수평하강.
[Objective] To investigate the expression of zebrafish vascular endothelial growth factor-2 (VEGFR-2) at different developmental stages. [Method] Total RNAs were extracted from 12, 24, 48, 72 and 96 hpf stage zebrafish embryos and larvae. Real-time quantitative RT-PCR was performed to examine the expression of VEGFR-2. The data were analyzed by 2-△△Ct method. [Result] The expression level of VEGFR-2 gene increased gradually from 12 to 72 hpf, and subsequently decreased at 96 hpf. The expression level was lowest at 12 hpf, highest at 72 hpf, and had significant differences when compared with that of other developmental stages. [Conclusion] The expression level of VEGFR-2 increases gradually before blood vessel maturation and decreases as blood vessels mature.