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C型利钠肽基因调控斑马鱼胚胎血管发育

张晶晶, 王新, 刘东

广东医科大学附属医院，湛江 524001；南通大学神经再生重点实验室，南通 226001

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:nGFP)，显微注射nppc特异性吗啉基反义寡核苷酸(morpholino)和nppc mRNA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。

关键词： nppc; 原位杂交 ; 血管发育 ; 斑马鱼

分类号：Q253

Natriuretic peptide precursor C coding gene contributes to zebrafish angiogenesis

ZHANG Jing-Jing, WANG XIN, LIU Dong

Affiliated Hospital of Guangdong Medical University, Zhanjiang 524001, China； Jiangsu Key Laboratory of Neuroregeneration, Nantong University, Nantong 226001, China

Abstract

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 of nppc in Tg (flk1:GFP) and Tg (fli1a:nGFP) transgenic lines. The morphology and endothelial 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.

Key words: nppc; whole mount in situ hybridization ; angiogenesis ; zebrafish

收稿日期：2016-07-29　　录用日期：2016-08-29

通讯作者：张晶晶,刘东　　E-mail: jingjing.zhang@live.com, liudongtom@gmail.com

引用本文：

张晶晶, 王新, 刘东. C型利钠肽基因调控斑马鱼胚胎血管发育[J]. 生理学报 2017; 69 (1): 11-16.

ZHANG Jing-Jing, WANG XIN, LIU Dong. Natriuretic peptide precursor C coding gene contributes to zebrafish angiogenesis. Acta Physiol Sin 2017; 69 (1): 11-16 (in Chinese with English abstract).