A Gata4 nuclear GFP transcriptional reporter to study endoderm and cardiac development in the mouse.

Claire S Simon,Nestor Saiz,Chen-Leng Cai,Tao Wu,Weibin Cai,Lu Zhang,Sonja Nowotschin,Anna-Katerina Hadjantonakis

doi:10.1242/bio.036517

Abstract

ABSTRACTThe GATA zinc-finger transcription factor GATA4 is expressed in a variety of tissues during mouse embryonic development and in adult organs. These include the primitive endoderm of the blastocyst, visceral endoderm of the early post-implantation embryo, as well as lateral plate mesoderm, developing heart, liver, lung and gonads. Here, we generate a novel Gata4 targeted allele used to generate both a Gata4H2B-GFP transcriptional reporter and a Gata4FLAG fusion protein to analyse dynamic expression domains. We demonstrate that the Gata4H2B-GFP transcriptional reporter faithfully recapitulates known sites of Gata4 mRNA expression and correlates with endogenous GATA4 protein levels. This reporter labels nuclei of Gata4 expressing cells and is suitable for time-lapse imaging and single cell analyses. As such, this Gata4H2B-GFP allele will be a useful tool for studying Gata4 expression and transcriptional regulation.This article has an associated First Person interview with the first author of the paper.

Highlights

GATA4 is a member of the highly conserved GATA family zinc-finger transcription factors (Viger et al, 2008), which bind to the GATA motif consensus sequence 5-AGATAG-3 in promoter and cis-regulatory elements of target genes to activate transcription (Arceci et al, 1993; Merika and Orkin, 1993)
While other primitive endoderm (PrE) markers GATA6, PDGFRA, SOX17 are expressed to some extent in uncommitted inner cell mass (ICM) cells, GATA4 is the earliest known PrE-specific marker (Artus et al, 2011; Plusa et al, 2008; Kurimoto et al, 2006; Arceci et al, 1993; Niakan et al, 2010; Morris et al, 2010; Chazaud et al, 2006; Ohnishi et al, 2014), making it a prime candidate reporter for the identification of PrE cells as they emerge in realtime
Generation of a Gata4H2B-GFP reporter To follow transcriptional activity of Gata4 in mouse embryonic development, we generated a bright nuclear localised fluorescent reporter that would be suitable for live imaging and for single-cell resolution analysis

Summary

Introduction

GATA4 is a member of the highly conserved GATA family zinc-finger transcription factors (Viger et al, 2008), which bind to the GATA motif consensus sequence 5-AGATAG-3 in promoter and cis-regulatory elements of target genes to activate transcription (Arceci et al, 1993; Merika and Orkin, 1993). GATA4 mutations are associated with a wide range of congenital heart defects including ventricular septal defects and gonadal anomalies (Garg et al, 2003; Pehlivan et al, 1999; Lourenco et al, 2011; Wang et al, 2011). Overexpression of either GATA4 or GATA6 is sufficient to induce differentiation of pluripotent embryonic stem (ES) cells to extra-embryonic endoderm like (XEN) cells (Fujikura et al, 2002; Schroter et al, 2015; Morrisey et al, 1996; Shimosato et al, 2007; Niakan et al, 2013), Gata is dispensable for PrE specification in vivo (Molkentin et al, 1997; Kuo et al, 1997)

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