A morpholino-based screen to identify novel genes involved in craniofacial morphogenesis

Dev Dyn. 2013 Jul;242(7):817-31. doi: 10.1002/dvdy.23969. Epub 2013 Jun 3.

Abstract

Background: The regulatory mechanisms underpinning facial development are conserved between diverse species. Therefore, results from model systems provide insight into the genetic causes of human craniofacial defects. Previously, we generated a comprehensive dataset examining gene expression during development and fusion of the mouse facial prominences. Here, we used this resource to identify genes that have dynamic expression patterns in the facial prominences, but for which only limited information exists concerning developmental function.

Results: This set of ∼80 genes was used for a high-throughput functional analysis in the zebrafish system using Morpholino gene knockdown technology. This screen revealed three classes of cranial cartilage phenotypes depending upon whether knockdown of the gene affected the neurocranium, viscerocranium, or both. The targeted genes that produced consistent phenotypes encoded proteins linked to transcription (meis1, meis2a, tshz2, vgll4l), signaling (pkdcc, vlk, macc1, wu:fb16h09), and extracellular matrix function (smoc2). The majority of these phenotypes were not altered by reduction of p53 levels, demonstrating that both p53-dependent and -independent mechanisms were involved in the craniofacial abnormalities.

Conclusions: This Morpholino-based screen highlights new genes involved in development of the zebrafish craniofacial skeleton with wider relevance to formation of the face in other species, particularly mouse and human.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • Calcium-Binding Proteins / genetics
  • Calcium-Binding Proteins / metabolism
  • Facial Bones / embryology*
  • Facial Bones / metabolism*
  • Gene Expression Regulation, Developmental / genetics
  • Gene Expression Regulation, Developmental / physiology
  • Homeodomain Proteins / genetics
  • Homeodomain Proteins / metabolism
  • Mice
  • Morphogenesis / genetics
  • Morphogenesis / physiology
  • Morpholinos
  • Myeloid Ecotropic Viral Integration Site 1 Protein
  • Neoplasm Proteins / genetics
  • Neoplasm Proteins / metabolism
  • Repressor Proteins / genetics
  • Repressor Proteins / metabolism
  • Skull / embryology*
  • Skull / metabolism*
  • Zebrafish
  • Zebrafish Proteins / genetics
  • Zebrafish Proteins / metabolism

Substances

  • Calcium-Binding Proteins
  • Homeodomain Proteins
  • MEIS1 protein, human
  • Meis1 protein, mouse
  • Morpholinos
  • Mrg1 protein, mouse
  • Myeloid Ecotropic Viral Integration Site 1 Protein
  • Neoplasm Proteins
  • Repressor Proteins
  • SMOC-2 protein, mouse
  • Tshz2 protein, mouse
  • Zebrafish Proteins