Transcriptome Analysis Did Not Show Endogenous Stem Cell Characteristics in Murine Lgr5+ Retinal Cells

Int J Mol Sci. 2019 Jul 19;20(14):3547. doi: 10.3390/ijms20143547.

Abstract

Lgr5, an intestinal adult stem cell marker, was recently also found in neuronal tissues. We investigated whether retinal Lgr5+ cells express properties of neural stem cells (NSC) and/or of differentiated interneurons during retinal development. RNA was isolated from Lgr5+ and Lgr5- populations from postnatal day 5 (PN5) and adult retinas of Lgr5EGFP-Ires-CreERT2 knock-in mice sorted by fluorescence-activated cell sorting (FACS). Transcriptome analyses were performed on two RNA samples of each developmental stage (PN5 and adult). The online platform PANTHER (Protein ANalysis THrough Evolutionary Relationships) was used to determine overrepresented gene ontology (GO) terms of biological processes within the set of differentially expressed genes. The detailed evaluation included gene expression in regard to stem cell maintenance/proliferation, cell cycle, and Wnt signaling but also markers of differentiated retinal neurons. None of the enriched GO terms of upregulated genes of Lgr5+ cells showed a positive association to NSC. On the contrary, NSC maintenance and proliferation rather prevail in the Lgr5- cell population. Furthermore, results suggesting that Wnt signaling is not active in the Lgr5+ population. Therefore, our transcriptome analysis of Lgr5+ retinal cells suggest that these cells are differentiated neurons, specifically glycinergic amacrine cells.

Keywords: Lgr5+ cells; amacrine cells; interneurons; retinal development; stem cells; transcriptomics.

MeSH terms

  • Amacrine Cells / metabolism
  • Animals
  • Cell Differentiation / genetics
  • Cell Proliferation
  • Female
  • Gene Expression Profiling*
  • Gene Expression Regulation, Developmental
  • Male
  • Mice
  • Neurons / metabolism
  • Receptors, G-Protein-Coupled / genetics*
  • Receptors, G-Protein-Coupled / metabolism
  • Retina / cytology*
  • Stem Cells / cytology*
  • Stem Cells / metabolism*
  • Transcriptome*

Substances

  • Lgr5 protein, mouse
  • Receptors, G-Protein-Coupled