摘要
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 investigate the expression of the natriuretic peptide precursor C coding gene nppc and its role in angiogenesis during embryonic period of the zebrafish. Whole mount in situ hybridization was performed to detect the expression pattern of nppc. nppc specific morpholino and nppc mRNA were injected respectively into the one-cell stage embryo to specifically knock-down and rescue the expression ofnppc in Tg （flkl：GFP） and Tg （flila：nGFP） transgenic lines. The morphology and endothe- lial cell number of intersegmental vessel （ISV） were analyzed after imaging using the laser scanning confocal microscope. The results revealed that nppc was expressed in the brain, heart and vasculature of zebrafish larvae at 24 and 48 hours post-fertilization （hpf）. Knock-down of nppc affected the development of ISV. Endothelial cell number was reduced after the knock-down of nppc. These results suggest that nppc controls zebrafish angiogenesis by affecting the endothelial cell proliferation and migration.
作者
张晶晶
王新
刘东
ZHANG Jlng-Jlng WANG Xin LIU Dong(Affiliated Hospital of Guangdong Medical University, Zhanjiang 524001, China Jiangsu Key Laboratory of Neuroregeneration Nantong University, Nantong 226001, China)
出处
《生理学报》
CAS
CSCD
北大核心
2017年第1期11-16,共6页
Acta Physiologica Sinica
基金
supported by grants from the National Natural Science Foundation of China(No.31370824)
the Special Support Plan of Excellent Young Talents
Guangdong Province
China(No.[2015]8)
Initial Program of Excellent Young Teachers in Colleges and Universities in Guangdong Province
China(No.[2016]6:YQ2015081)
关键词
nppc:原位杂交
血管发育
斑马鱼
nppc
whole mount in situ hybridization
angiogenesis
zebrafish
