Equarin is involved as an FGF signaling modulator in chick lens differentiation

Dev Biol. 2012 Aug 1;368(1):109-17. doi: 10.1016/j.ydbio.2012.05.029. Epub 2012 May 31.

Abstract

Lens growth involves the proliferation of epithelial cells, followed by their migration to the equator region and differentiation into secondary fiber cells. It is widely accepted that fibroblast growth factor (FGF) signaling is required for the differentiation of lens epithelial cells into crystallin-rich fibers, but this signaling is insufficient to induce full differentiation. To better understand lens development, investigatory and functional analyses of novel molecules are required. Here, we demonstrate that Equarin, which is a novel secreted molecule, was expressed exclusively in the lens equator region during chick lens development. Equarin upregulated the expression of fiber markers, as demonstrated using in ovo electroporation. In a primary lens cell culture, Equarin promoted the biochemical and morphological changes associated with the differentiation of lens epithelial cells to fibers. A loss-of-function analysis was performed using zinc-finger nucleases targeting the Equarin gene. Lens cell differentiation was markedly inhibited when endogenous Equarin was blocked, indicating that Equarin was essential for normal chick lens differentiation. Furthermore, biochemical analysis showed that Equarin directly bound to FGFs and heparan sulfate proteoglycan and thereby upregulated the expression of phospho-ERK1/2 (ERK-P) proteins, the downstream of the FGF signaling pathway, in vivo and in vitro. Conversely, the absence of endogenous Equarin clearly diminished FGF-induced fiber differentiation. Taken together, our results suggest that Equarin is involved as an FGF modulator in chick lens differentiation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Avian Proteins / genetics
  • Avian Proteins / metabolism*
  • Base Sequence
  • Blotting, Western
  • COS Cells
  • Cell Differentiation
  • Cells, Cultured
  • Chick Embryo
  • Chickens
  • Chlorocebus aethiops
  • Eye Proteins / genetics
  • Eye Proteins / metabolism*
  • Fibroblast Growth Factors / genetics
  • Fibroblast Growth Factors / metabolism*
  • Gene Expression Regulation, Developmental
  • Heparan Sulfate Proteoglycans / metabolism
  • Immunohistochemistry
  • In Situ Hybridization
  • Lens, Crystalline / cytology
  • Lens, Crystalline / embryology
  • Lens, Crystalline / metabolism*
  • Molecular Sequence Data
  • Mutation
  • Protein Binding
  • Sequence Homology, Nucleic Acid
  • Signal Transduction*
  • beta-Crystallins / metabolism

Substances

  • Avian Proteins
  • Eye Proteins
  • Heparan Sulfate Proteoglycans
  • beta-Crystallins
  • Fibroblast Growth Factors