PUBLICATION

Protein-Trap Insertional Mutagenesis Uncovers New Genes Involved in Zebrafish Skin Development, Including a Neuregulin 2a-Based ErbB Signaling Pathway Required during Median Fin Fold Morphogenesis

Authors
Westcot, S.E., Hatzold, J., Urban, M.D., Richetti, S.K., Skuster, K.J., Harm, R.M., Lopez Cervera, R., Umemoto, N., McNulty, M.S., Clark, K.J., Hammerschmidt, M., Ekker, S.C.
ID
ZDB-PUB-150626-2
Date
2015
Source
PLoS One   10: e0130688 (Journal)
Registered Authors
Ekker, Stephen C., Hammerschmidt, Matthias, Harm, Rhianna, Hatzold, Julia, McNulty, Melissa, Richetti, Stefânia, Urban, Mark, Westcot, Stephanie
Keywords
none
MeSH Terms
  • Animals
  • Neuregulins/genetics
  • Neuregulins/metabolism*
  • Skin/embryology*
  • Skin/metabolism
  • Zebrafish/embryology*
  • Zebrafish/genetics
  • Zebrafish/metabolism
  • Mutagenesis, Insertional
  • Alleles
  • Zebrafish Proteins/genetics
  • Zebrafish Proteins/metabolism*
  • Oncogene Proteins v-erbB/genetics
  • Oncogene Proteins v-erbB/metabolism*
  • Organogenesis/genetics*
  • Proto-Oncogene Proteins c-akt/metabolism
  • Animal Fins/embryology*
  • Animal Fins/metabolism
  • Signal Transduction/genetics
  • Gene Expression Regulation, Developmental
(all 20)
PubMed
26110643 Full text @ PLoS One
Abstract
Skin disorders are widespread, but available treatments are limited. A more comprehensive understanding of skin development mechanisms will drive identification of new treatment targets and modalities. Here we report the Zebrafish Integument Project (ZIP), an expression-driven platform for identifying new skin genes and phenotypes in the vertebrate model Danio rerio (zebrafish). In vivo selection for skin-specific expression of gene-break transposon (GBT) mutant lines identified eleven new, revertible GBT alleles of genes involved in skin development. Eight genes-fras1, grip1, hmcn1, msxc, col4a4, ahnak, capn12, and nrg2a-had been described in an integumentary context to varying degrees, while arhgef25b, fkbp10b, and megf6a emerged as novel skin genes. Embryos homozygous for a GBT insertion within neuregulin 2a (nrg2a) revealed a novel requirement for a Neuregulin 2a (Nrg2a) - ErbB2/3 - AKT signaling pathway governing the apicobasal organization of a subset of epidermal cells during median fin fold (MFF) morphogenesis. In nrg2a mutant larvae, the basal keratinocytes within the apical MFF, known as ridge cells, displayed reduced pAKT levels as well as reduced apical domains and exaggerated basolateral domains. Those defects compromised proper ridge cell elongation into a flattened epithelial morphology, resulting in thickened MFF edges. Pharmacological inhibition verified that Nrg2a signals through the ErbB receptor tyrosine kinase network. Moreover, knockdown of the epithelial polarity regulator and tumor suppressor lgl2 ameliorated the nrg2a mutant phenotype. Identifying Lgl2 as an antagonist of Nrg2a - ErbB signaling revealed a significantly earlier role for Lgl2 during epidermal morphogenesis than has been described to date. Furthermore, our findings demonstrated that successive, coordinated ridge cell shape changes drive apical MFF development, making MFF ridge cells a valuable model for investigating how the coordinated regulation of cell polarity and cell shape changes serves as a crucial mechanism of epithelial morphogenesis.
Genes / Markers
Figures
Figure Gallery (14 images) / 2
Show all Figures
Expression
Phenotype
Mutations / Transgenics
Allele Construct Type Affected Genomic Region
gz7TgTransgenic Insertion
    mn0078GtTransgenic Insertion
    mn0156GtTransgenic Insertion
    mn0175GtTransgenic Insertion
    mn0196GtTransgenic Insertion
    mn0237GtTransgenic Insertion
    mn0245GtTransgenic Insertion
    mn0261GtTransgenic Insertion
    mn0263GtTransgenic Insertion
    mn0275GtTransgenic Insertion
    1 - 10 of 15
    Show
    Human Disease / Model
    No data available
    Sequence Targeting Reagents
    Target Reagent Reagent Type
    llgl2MO1-llgl2MRPHLNO
    tp53MO4-tp53MRPHLNO
    1 - 2 of 2
    Show
    Fish
    Antibodies
    Name Type Antigen Genes Isotypes Host Organism
    Ab1-col2amonoclonal
      IgG1Mouse
      Ab1-tp63monoclonalIgG2aMouse
      Ab5-aktmonoclonal
        Rabbit
        Ab26-mapkmonoclonal
          IgG1Mouse
          1 - 4 of 4
          Show
          Orthology
          No data available
          Engineered Foreign Genes
          Marker Marker Type Name
          EGFPEFGEGFP
          GFPEFGGFP
          mRFP1EFGmRFP1
          1 - 3 of 3
          Show
          Mapping
          No data available