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本研究旨在探索C型利钠肽前体蛋白基因(natriuretic peptide precursor C,nppc)在斑马鱼胚胎期的表达及其在血管发育过程中的功能。利用胚胎整体原位杂交技术检测nppc基因在斑马鱼胚胎期的表达。同时使用转基因斑马鱼系Tg(flk1:GFP)和Tg(fli1a:n GFP),显微注射nppc特异性吗啉基反义寡核苷酸(morpholino)和nppc m RNA调控nppc基因表达,利用激光共聚焦显微镜观察分析斑马鱼节间血管(intersegment vessel,ISV)表型,并统计内皮细胞数目。结果显示,在受精后24 h和48 h,nppc基因在斑马鱼脑、心脏、血管系统中都有表达。下调nppc基因表达导致ISV发育缺陷,ISV内皮细胞数目减少。以上结果表明下调nppc基因表达可能通过抑制内皮细胞增殖和内皮细胞迁移调控斑马鱼胚胎血管发育。
This study aimed to explore the expression of natriuretic peptide precursor C (nppc) during zebrafish embryogenesis and its function during vascular development. Detection of nppc gene expression in zebrafish embryos by whole - mount in situ hybridization. At the same time, the expression of nppc gene was regulated by microinjection of nppc-specific morpholino-morpholino and nppc-m RNA into the transgenic zebrafish lines Tg (flk1: GFP) and Tg (fli1a: n GFP) Confocal microscopy was used to analyze the phenotypes of the segments of the testes (ISV), and the number of endothelial cells was counted. The results showed that nppc gene was expressed in brain, heart and blood system of zebrafish at 24 h and 48 h after fertilization. Down-regulation of nppc gene expression leads to defects in ISV development and reduction of ISV endothelial cells. The above results show that down-regulation of nppc gene expression may regulate zebrafish embryonic vascular development by inhibiting endothelial cell proliferation and endothelial cell migration.