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

Authors

  • Karolina Mizeracka,

    1. Department of Genetics, Department of Ophthalmology, Harvard Medical School, Boston, Massachusetts
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  • Jeffrey M. Trimarchi,

    1. Department of Genetics, Department of Ophthalmology, Harvard Medical School, Boston, Massachusetts
    Current affiliation:
    1. Department of Genetics, Development and Cell Biology, Iowa State University, Ames, IA
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  • Michael B. Stadler,

    1. Friedrich Miescher Institute for Biomedical Research, Basel, Switzerland
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  • Constance L. Cepko

    Corresponding author
    1. Department of Genetics, Department of Ophthalmology, Harvard Medical School, Boston, Massachusetts
    2. Howard Hughes Medical Institute, Department of Genetics, Department of Ophthalmology, Harvard Medical School, Boston, Massachusetts
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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. Developmental Dynamics, 242:1147–1159, 2013. © 2013 Wiley Periodicals, Inc.

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