Clec14a基因调控斑马鱼胚胎血管发育  

Clec14a gene contributes to zebrafish angiogenesis

作　　者：杨晋娴 王新[3] 刘东[3] 张海骏 邵义祥 YANG Jinxian;WANG xin;LIU Dong;ZHANG Haijun;SHAO Yixiang(Xinglin College of Nantong University, Nantong University, Jiangsu 226001;Laboratory Animal Center, Nantong University, Jiangsu 226001;Jiangsu Key Laboratory of Neuroregeneration, Nantong University, Jiangsu 226001)

机构地区：[1]南通大学杏林学院,江苏226001 [2]南通大学实验动物中心,江苏226001 [3]南通大学江苏省神经再生重点实验室,江苏226001

出　　处：《交通医学》2018年第6期535-539,共5页Medical Journal of Communications

基　　金：国家自然科学基金项目(81870359);江苏省研究生科研创新项目(KYCX17-1909);南通大学杏林学院科研基金项目(2016K138)

摘　　要：目的:探索Clec14a在斑马鱼胚胎期的表达及其在血管发育过程中的功能,探讨该基因调控血管发育机制。方法:通过胚胎整体原位杂交技术,观察Clec14a基因在斑马鱼胚胎期的表达情况;在转基因斑马鱼系Tg(kdrl:mCherry)和Tg(fli1a:nGFP)中,显微注射吗啉修饰的反义寡核苷酸(morpholinoantisense oligos)下调Clec14a基因表达,同时在转基因斑马鱼系Tg(kdrl:mCherry)中注射Clec14a-mRNA调控Clec14a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,统计分析内皮细胞迁移速度和数目的变化,并应用qRT-PCR技术检测下调后该基因mRNA表达情况。结果:在受精后24 h、48 h和72 h,Clec14a基因在斑马鱼脑、心脏、血管系统中有表达;Clec14a基因表达下调导致ISV发育缺陷,内皮细胞数目减少,抑制其迁移速度。结论:下调Clec14a基因表达能够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。Objective:To investigate the expression of Clec14 a and its role in angiogenesis during embryonic period of the zebrafish. Methods:Whole mount in situ hybridization was performed to detect the expression pattern of Clec14 a.Clec14 aspecificmorpholino was injected respectively into the one-cell stage embryo to specifically knock-down expression of Clec14 a in Tg(kdrl:mCherry) and Tg(fli1 a:nGFP) transgenic lines. And Clec14 amRNA was injected respectively into the embryo to rescue the expression of Clec14 a in Tg(kdrl:mCherry). The morphology and endothelial cell number of intersegmental vessel(ISV) were analyzed after imaging by using the laser scanning confocal microscope. And qRT-PCR technology was used to detect the mRNA expression after the knock-down of Clec14 a. Results:Clec14 a was expressed in the brain, heart and vasculature of zebrafish larvae at 24, 48 and 72 hours post-fertilization(hpf). Knock-down of Clec14 a affected the development of ISV. Endothelial cell number was reduced after the knock-down of Clec14 a. Conclusions:Clec14 a controls zebrafish angiogenesis by affecting the endothelial cell proliferation and migration.

关 键 词：Clec14a 血管发育 原位杂交 斑马鱼 

分 类 号：Q75[生物学—分子生物学]