PUBLICATION

An evolutionarily conserved kernel of gata5, gata6, otx2 and prdm1a operates in the formation of endoderm in zebrafish

Authors

Tseng, W.F., Jang, T.H., Huang, C.B., and Yuh, C.H.

ID

ZDB-PUB-110719-20

Date

2011

Source

Developmental Biology 357(2): 541-57 (Journal)

Registered Authors

Yuh, Chiou-Hwa (Cathy)

Keywords

kernel, mesendoderm, zebrafish

MeSH Terms

Oligonucleotides, Antisense/pharmacology
DNA Mutational Analysis
Endoderm/drug effects
Endoderm/embryology*
Endoderm/metabolism
RNA, Messenger/genetics
RNA, Messenger/metabolism
Conserved Sequence/genetics*
GATA5 Transcription Factor/genetics
GATA5 Transcription Factor/metabolism
Genetic Loci/genetics
Gene Regulatory Networks/genetics*
Protein Binding
Epistasis, Genetic/drug effects
Molecular Sequence Data
Base Sequence
DNA-Binding Proteins/genetics
DNA-Binding Proteins/metabolism
Gene Expression Regulation, Developmental/drug effects
Otx Transcription Factors/genetics
Otx Transcription Factors/metabolism
Nuclear Proteins/genetics
Nuclear Proteins/metabolism
GATA Transcription Factors/genetics
GATA Transcription Factors/metabolism
Evolution, Molecular*
Zebrafish/embryology*
Zebrafish/genetics*
Reproducibility of Results
Zebrafish Proteins/genetics
Zebrafish Proteins/metabolism*
Time Factors
Gene Expression Profiling
Animals
Body Patterning/drug effects
Body Patterning/genetics

(all 36)

PubMed

21756893 Full text @ Dev. Biol.

Abstract

An evolutionarily conserved subcircuit (kernel) dedicated to a specific developmental function is found at the top of the gene regulatory networks (GRNs) hierarchy. Here we comprehensively demonstrate that a pan-deuterostome endoderm specification kernel exists in zebrafish. We analyzed interactions among gata5, gata6, otx2 and prdm1a using specific morpholino knockdowns and measured the gene expression profiles by quantitative real-time RT-PCR and in situ hybridization. The mRNA rescue experiment validated the specificity of the morpholino knockdown. We found that the interactions among gata5, gata6, otx2 and prdm1a determine the initial specification of the zebrafish endoderm. Although otx2 can activate both gata5 and gata6, and the prdm1a/krox homologue also activates some endoderm transcription factors, a feedback loop from Gata to otx2 and prdm1a is missing. Furthermore, we found the positive regulation between gata5 and gata6 to further lock-on the mesendoderm specification by the Gata family. Chromatin immunoprecipitation was used to further validate the recruitment of Otx2 to the gata5 and gata6 loci. Functional assays revealed that module B of gata6 and the basal promoter of gata5 drive the gene at the mesendoderm, and mutational analysis demonstrated that Otx2 and Gata5/6 contribute to reporter gene activation. This is the first direct evidence for evolutionarily conserved endoderm specification across echinoderms and vertebrates.

Genes / Markers

Figures