PUBLICATION

Grainyhead-like 3 regulation of endothelin-1 in the pharyngeal endoderm is critical for growth and development of the craniofacial skeleton

Authors
Dworkin, S., Simkin, J., Darido, C., Partridge, D.D., Georgy, S.R., Caddy, J., Wilanowski, T., Lieschke, G.J., Doggett, K., Heath, J.K., Jane, S.M.
ID
ZDB-PUB-140611-1
Date
2014
Source
Mechanisms of Development   133: 77-90 (Journal)
Registered Authors
Doggett, Karen, Dworkin, Seb, Heath, Joan K., Jane, Stephen M., Lieschke, Graham J.
Keywords
Craniofacial defects, Endoderm, Endothelin-1, Grainyhead-like, Pharynx, grhl3
MeSH Terms
  • Animals
  • Animals, Genetically Modified
  • Binding Sites/genetics
  • Chromatin Immunoprecipitation
  • Endothelin-1/genetics*
  • Endothelin-1/metabolism
  • Epistasis, Genetic
  • Facial Bones/growth & development
  • Facial Bones/metabolism
  • Gene Expression Regulation, Developmental
  • Gene Knockdown Techniques
  • Humans
  • Models, Biological
  • Neural Crest/cytology
  • Neural Crest/metabolism
  • RNA, Messenger/genetics
  • RNA, Messenger/metabolism
  • Regulatory Sequences, Nucleic Acid
  • Signal Transduction
  • Skull/growth & development
  • Skull/metabolism
  • Zebrafish/genetics*
  • Zebrafish/growth & development*
  • Zebrafish/metabolism
  • Zebrafish Proteins/antagonists & inhibitors
  • Zebrafish Proteins/genetics*
  • Zebrafish Proteins/metabolism
PubMed
24915580 Full text @ Mech. Dev.
Abstract
Craniofacial development is a highly conserved process that requires complex interactions between neural crest cells (NCCs) and pharyngeal tissues derived from all three germ layers. Signals emanating from the pharyngeal endoderm drive differentiation of NCCs into craniofacial cartilage, and disruption of this process underpins several human craniofacial defects (CFD). Here, we demonstrate that morpholino (MO)-mediated knockdown in zebrafish of the highly conserved transcription factor grainyhead-like 3 (grhl3), which is selectively expressed in the pharyngeal endoderm, leads to severe hypoplasia of the lower jaw cartilages. Phylogenetic analysis of conserved grhl-binding sites in gene regulatory regions identified endothelin-1 (edn1) as a putative direct grhl3 target gene, and this was confirmed by chromatin precipitation (ChIP) assays in zebrafish embryos. Injection of sub-phenotypic concentrations of MOs targeting both grhl3 and edn1 induced jaw abnormalities, and injection of edn1 mRNA into grhl3-morphants rescued both pharyngeal expression of the downstream effectors of edn1, and jaw cartilage formation. This study sheds new light on the role of endodermal endothelin-1 in vertebrate jaw development, and highlights potential new genetic defects that could underpin human CFD.
Genes / Markers
Figures
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Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Antibodies
Orthology
Engineered Foreign Genes
Mapping