Premium
Isolation of a ventricle‐specific promoter for the zebrafish ventricular myosin heavy chain ( vmhc ) gene and its regulation by GATA factors during embryonic heart development
Author(s) -
Park JaeSun,
Kim HyungSeok,
Kim JunDae,
Seo Jungwon,
Chung KyungSook,
Lee HoSa,
Huh TaeLin,
Jo Inho,
Kim YongOu
Publication year - 2009
Publication title -
developmental dynamics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.634
H-Index - 141
eISSN - 1097-0177
pISSN - 1058-8388
DOI - 10.1002/dvdy.21964
Subject(s) - biology , zebrafish , gene knockdown , gata4 , myosin , microbiology and biotechnology , promoter , heart development , gene , gene expression , green fluorescent protein , genetics , embryonic stem cell
We investigated chamber‐specific gene expression by isolating a 2.2‐kb polymerase chain reaction product containing the 5′‐flanking region of the zebrafish ventricular myosin heavy‐chain gene ( vmhc ). Promoter analysis revealed that the fragment, consisting of nucleotides from −301 to +26, is sufficient for vmhc promoter activity. Among several putative cis ‐acting elements in the region, a GATA‐binding site was identified to be crucial for the activity of the promoter, as evidenced by the complete abolishment of promoter activity by a single nucleotide substitution of GATA‐binding site (−287, C→T). Knockdown of GATA‐binding proteins 4 and 6 (GATA4 and ‐6) by their antisense morpholino oligonucleotides resulted in reduced green fluorescent protein (GFP) reporter gene and endogenous vmhc expression. These findings suggest that GATA4 and ‐6 play a key role in the regulation of vmhc expression in the ventricle. In addition, the vmhc promoter and the transgenic zebrafish ( vmhc:gfp ) are useful tools to study the formation and function of the ventricle. Developmental Dynamics 238:1574–1581, 2009. © 2009 Wiley‐Liss, Inc.