Transient and transgenic analysis of the zebrafish ventricular myosin heavy chain (vmhc) promoter: An inhibitory mechanism of ventricle‐specific gene expression
Open Access
- 25 March 2009
- journal article
- research article
- Published by Wiley in Developmental Dynamics
- Vol. 238 (6), 1564-1573
- https://doi.org/10.1002/dvdy.21929
Abstract
The zebrafish ventricular myosin heavy chain (vmhc) gene exhibits restricted expression in the ventricle. However, the molecular mechanism underlying this chamber‐specific expression is unclear. Here, we exploited both transient and transgenic technologies to dissect the zebrafish vmhc promoter. We demonstrated that a combination of two transient assays in this animal model quickly identified chamber‐specific cis‐elements, isolating a 2.2 kb fragment upstream from the vmhc gene that can drive ventricle‐specific expression. Furthermore, deletion analysis identified multiple cis‐elements that exhibited cardiac‐specific expression. To achieve chamber specificity, a distal element was required to coordinate with and suppress a proximal enhancer element. Finally, we discovered that Nkx2.5‐binding sites (NKE) were essential for this repressive function. In summary, our study of the zebrafish vmhc promoter suggests that ventricle‐specific expression is achieved through an inhibitory mechanism that suppresses expression in the atrium. Developmental Dynamics 238:1564–1573, 2009.Keywords
This publication has 50 references indexed in Scilit:
- Nkx genes regulate heart tube extension and exert differential effects on ventricular and atrial cell numberDevelopmental Biology, 2008
- Depletion of zebrafish essential and regulatory myosin light chains reduces cardiac function through distinct mechanismsCardiovascular Research, 2008
- The Tol2kit: A multisite gateway‐based construction kit for Tol2 transposon transgenesis constructsDevelopmental Dynamics, 2007
- Vessel and Blood Specification Override Cardiac Potential in Anterior MesodermDevelopmental Cell, 2007
- Depletion of Zebrafish Titin Reduces Cardiac Contractility by Disrupting the Assembly of Z-Discs and A-BandsCirculation Research, 2007
- Transposons as tools for enhancer trap screens in vertebratesGenome Biology, 2007
- Tol2: a versatile gene transfer vector in vertebratesGenome Biology, 2007
- Evaluating the biological relevance of putative enhancers using Tol2 transposon-mediated transgenesis in zebrafishNature Protocols, 2006
- Single-stranded DNA-binding Proteins PURα and PURβ Bind to a Purine-rich Negative Regulatory Element of the α-Myosin Heavy Chain Gene and Control Transcriptional and Translational Regulation of the Gene ExpressionPublished by Elsevier BV ,2003
- Prediction of complete gene structures in human genomic DNAJournal of Molecular Biology, 1997