Analysis of gene expression in wild-type and Notch1 mutant retinal cells by single cell profiling

Dev Dyn. 2013 Oct;242(10):1147-59. doi: 10.1002/dvdy.24006. Epub 2013 Sep 2.

Abstract

Background: The vertebrate retina comprises sensory neurons, the photoreceptors, as well as many other types of neurons and one type of glial cell. These cells are generated by multipotent and restricted retinal progenitor cells (RPCs), which express Notch1. Loss of Notch1 in RPCs late during retinal development results in the overproduction of rod photoreceptors at the expense of interneurons and glia.

Results: To examine the molecular underpinnings of this observation, microarray analysis of single retinal cells from wild-type or Notch1 conditional knockout retinas was performed. In situ hybridization was carried out to validate some of the findings.

Conclusions: The majority of Notch1-mutant cells lost expression of known Notch target genes. These cells also had low levels of RPC and cell cycle genes, and robustly up-regulated rod precursor genes. In addition, single wild-type cells, in which cell cycle marker genes were down-regulated, expressed markers of both rod photoreceptors and interneurons.

Keywords: Notch; cell fate; microarray; progenitor; retina.

Publication types

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

MeSH terms

  • Animals
  • Cell Cycle / physiology*
  • Gene Expression Profiling
  • Gene Expression Regulation, Developmental / physiology*
  • Gene Knockdown Techniques
  • Mice
  • Mutation
  • Oligonucleotide Array Sequence Analysis
  • Receptor, Notch1 / genetics
  • Receptor, Notch1 / metabolism*
  • Retinal Rod Photoreceptor Cells / cytology
  • Retinal Rod Photoreceptor Cells / metabolism*
  • Stem Cells / cytology
  • Stem Cells / metabolism*

Substances

  • Notch1 protein, mouse
  • Receptor, Notch1