Inhibition of zebrafish epidermal growth factor receptor activity results in cardiovascular defects
Inhibition of zebrafish epidermal growth factor receptor activity results in cardiovascular defects
复制标题
DOI：
10.1016/s0925-4773(03)00068-6
复制
发表时间：
2003-07-01
影响因子：
2.6
通讯作者：
Klagsbrun, M
中科院分区：
文献类型：
--
作者：
Goishi, K;Lee, P;Klagsbrun, M
任何表皮生长因子（EGF）受体酪氨酸激酶的生理作用在斑马鱼中尚未确定。我们分离出一个斑马鱼同源EGFR（EGFR），显示了63%的氨基酸的整体身份，人类EGFR，但与90%的氨基酸的激酶结构域的一致性。整片原位杂交显示egfr转录本在原肠胚形成、体节发生及后期普遍存在。在中国仓鼠卵巢细胞中表达的斑马鱼Egfr是一种功能性受体，通过受体酪氨酸磷酸化和MAP激酶激活对EGF产生应答。通过使用EGFR激酶抑制剂和反义吗啉代（MO）抑制其活性来确定斑马鱼体内EGFR的功能，所述EGFR激酶抑制剂和反义吗啉代（MO）分别抑制EGFR激酶活性和EGFR信使RNA向蛋白质的翻译。斑马鱼是研究心血管发育的一个特别好的模型，因为斑马鱼是透明的，可以直接观察心脏和血管中的循环。抑制斑马鱼Egfr活性在体内阻碍了血流通过流出道进入主动脉，并阻碍了循环中的轴向和intersegmentated血管受精后80小时。心脏分析显示，心腔和心包囊扩张，流出道变窄。总之，这些结果表明，斑马鱼Egfr在发育中的斑马鱼中具有正常循环所需的心血管功能。(C)2003爱思唯尔爱尔兰有限公司保留所有权利。
The physiological role of any of the epidermal growth factor (EGF) receptor tyrosine kinases has yet to be determined in zebrafish. We isolated a zebrafish homologue of EGFR (egfr) that shows a 63% amino acid overall identity to human EGFR but with 90% amino acid identity in the kinase domain. Whole mount in situ hybridization showed ubiquitous distribution of egfr transcripts during gastrulation, somitogenesis and later stages. When expressed in Chinese hamster ovary cells, zebrafish Egfr was a functional receptor that responded to EGF by receptor tyrosine phosphorylation and activation of MAP kinase. The function of zebrafish Egfr in vivo was determined by inhibiting its activity using EGFR kinase inhibitors and antisense morpholinos (MO), which inhibited Egfr kinase activity and translation of egfr messenger RNA into protein, respectively. The zebrafish is a particularly excellent model for studying cardiovascular development because zebrafish are transparent allowing direct visualization of the heart and circulation in the blood vessels. Inhibition of zebrafish Egfr activity in vivo impeded blood flow via the outflow tract into the aorta and impeded circulation in the axial and intersegmental vessels by 80 h post-fertilization. Analysis of the heart showed that the heart chambers and pericardial sacs were dilated and the outflow tracts were narrowed. Together these results suggested that zebrafish Egfr has a cardiovascular function in the developing zebrafish that is required for normal circulation. (C) 2003 Elsevier Ireland Ltd. All rights reserved.