Etsrp/Etv2 is directly regulated by Foxc1a/b in the zebrafish angioblast. Circ Res 2012 Jan 20;110(2):220-9
Date
12/03/2011Pubmed ID
22135404Pubmed Central ID
PMC3457812DOI
10.1161/CIRCRESAHA.111.251298Scopus ID
2-s2.0-84856043722 (requires institutional sign-in at Scopus site) 45 CitationsAbstract
RATIONALE: Endothelial cells are developmentally derived from angioblasts specified in the mesodermal germ cell layer. The transcription factor etsrp/etv2 is at the top of the known genetic hierarchy for angioblast development. The transcriptional events that induce etsrp expression and angioblast specification are not well understood.
OBJECTIVE: We generated etsrp:gfp transgenic zebrafish and used them to identify regulatory regions and transcription factors critical for etsrp expression and angioblast specification from mesoderm.
METHODS AND RESULTS: To investigate the mechanisms that initiate angioblast cell transcription during embryogenesis, we have performed promoter analysis of the etsrp locus in zebrafish. We describe three enhancer elements sufficient for endothelial gene expression when place in front of a heterologous promoter. The deletion of all 3 regulatory regions led to a near complete loss of endothelial expression from the etsrp promoter. One of the enhancers, located 2.3 kb upstream of etsrp contains a consensus FOX binding site that binds Foxc1a and Foxc1b in vitro by EMSA and in vivo using ChIP. Combined knockdown of foxc1a/b, using morpholinos, led to a significant decrease in etsrp expression at early developmental stages as measured by quantitative reverse transcriptase-polymerase chain reaction and in situ hybridization. Decreased expression of primitive erythrocyte genes scl and gata1 was also observed, whereas pronephric gene pax2a was relatively normal in expression level and pattern.
CONCLUSIONS: These findings identify mesodermal foxc1a/b as a direct upstream regulator of etsrp in angioblasts. This establishes a new molecular link in the process of mesoderm specification into angioblast.
Author List
Veldman MB, Lin SAuthor
Matthew B. Veldman PhD Assistant Professor in the Cell Biology, Neurobiology and Anatomy department at Medical College of WisconsinMESH terms used to index this publication - Major topics in bold
AnimalsAnimals, Genetically Modified
Base Sequence
Binding Sites
Cell Lineage
Chromatin Immunoprecipitation
Electrophoretic Mobility Shift Assay
Embryo, Nonmammalian
Embryonic Stem Cells
Endothelial Cells
Forkhead Transcription Factors
Gene Expression Regulation, Developmental
Gene Knockdown Techniques
In Situ Hybridization
Mesoderm
Microscopy, Fluorescence
Molecular Sequence Data
Promoter Regions, Genetic
Recombinant Fusion Proteins
Reverse Transcriptase Polymerase Chain Reaction
Transcription, Genetic
Zebrafish
Zebrafish Proteins
