Inhibition of zebrafish epidermal growth factor receptor activity results in cardiovascular defects.
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.
Goishi
Katsutoshi
K
Vascular Biology Program, Children's Hospital and Harvard Medical School, Boston, MA 02115, USA.
Lee
Percy
P
Davidson
Alan J
AJ
Nishi
Eiichiro
E
Zon
Leonard I
LI
Klagsbrun
Michael
M
D000595:Amino Acid Sequence; D000818:Animals; D018376:Cardiovascular Abnormalities / Q000209:etiology / Q000378:metabolism*; D066246:ErbB Receptors / Q000037:antagonists & inhibitors / Q000378:metabolism*; D006801:Humans; D008969:Molecular Sequence Data; D011743:Pyrimidines / Q000494:pharmacology; D011758:Pyrroles / Q000494:pharmacology; D011799:Quinazolines; D051381:Rats; D020032:Tyrphostins / Q000494:pharmacology; D015027:Zebrafish / Q000196:embryology* / Q000378:metabolism
D011743:Pyrimidines; D011758:Pyrroles; D011799:Quinazolines; D020032:Tyrphostins; C101044:RTKI cpd; C410026:PKI 166; D066246:ErbB Receptors