Multiple intrinsic factors act in concert with Lhx2 to direct retinal gliogenesis

Sci Rep. 2016 Sep 8:6:32757. doi: 10.1038/srep32757.

Abstract

Müller glia (MG) are the principal glial cell type in the vertebrate retina. Recent work has identified the LIM homeodomain factor encoding gene Lhx2 as necessary for both Notch signaling and MG differentiation in late-stage retinal progenitor cells (RPCs). However, the extent to which Lhx2 interacts with other intrinsic regulators of MG differentiation is unclear. We investigated this question by investigating the effects of overexpression of multiple transcriptional regulators that are either known or hypothesized to control MG formation, in both wildtype and Lhx2-deficient RPCs. We observe that constitutively elevated Notch signaling, induced by N1ICD electroporation, inhibited gliogenesis in wildtype animals, but rescued MG development in Lhx2-deficient retinas. Electroporation of Nfia promoted the formation of cells with MG-like radial morphology, but did not drive expression of MG molecular markers. Plagl1 and Sox9 did not induce gliogenesis in wildtype animals, but nonetheless activated expression of the Müller marker P27(Kip1) in Lhx2-deficient cells. Finally, Sox2, Sox8, and Sox9 promoted amacrine cell formation in Lhx2-deficient cells, but not in wildtype retinas. These findings demonstrate that overexpression of individual gliogenic factors typically regulates only a subset of characteristic MG markers, and that these effects are differentially modulated by Lhx2.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Cycle Proteins / genetics
  • Cell Cycle Proteins / metabolism
  • Cell Differentiation / genetics
  • Electroporation
  • Eye Proteins / genetics
  • Eye Proteins / metabolism
  • Female
  • Gene Expression Regulation
  • Genes, Tumor Suppressor
  • Homeodomain Proteins / genetics
  • Homeodomain Proteins / metabolism
  • LIM-Homeodomain Proteins / genetics
  • LIM-Homeodomain Proteins / metabolism*
  • Male
  • Mice, Inbred Strains
  • Mice, Knockout
  • NFI Transcription Factors / genetics
  • NFI Transcription Factors / metabolism
  • Neuroepithelial Cells / physiology
  • Neuroglia / cytology
  • Neuroglia / physiology*
  • Retina / cytology*
  • Retina / metabolism
  • SOX Transcription Factors / genetics
  • SOX Transcription Factors / metabolism
  • Transcription Factors / genetics
  • Transcription Factors / metabolism*

Substances

  • Cell Cycle Proteins
  • Eye Proteins
  • Homeodomain Proteins
  • LIM-Homeodomain Proteins
  • Lhx2 protein, mouse
  • NFI Transcription Factors
  • Nfia protein, mouse
  • Plagl1 protein, mouse
  • Rax protein, mouse
  • SOX Transcription Factors
  • Transcription Factors