Unknown,Transcriptomics,Genomics,Proteomics

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Analysis of gene expression in wildtype and Notch1 mutant retinal cells by single cell profiling

ABSTRACT: Loss of Notch1 in retinal progenitor cells (RPCs) during postnatal retinal development results in the overproduction of rod photoreceptors at the expense of interneurons and glia. To examine the molecular underpinnings of this observation, microarray analysis of singla retinal cells from wildtype (WT) or Notch1 conditional knockout (N1-CKO) retinas was performed. The majority of N1-CKO cells lost expression of known Notch target genes. These cells also had low levels of RPC and cell cycle genes, and robustly upregulated rod precursor genes. In addition, single WT cells, in which cell cycle marker genes were downregulated, expressed markers of both rod photoreceptors and interneurons. These results demonstrate that individual, newly postmitotic retinal cells can begin to differentiate into more than one cell type, and that this transitional state may be dependent on Notch1 signaling. We used microarrays to examine gene expression changes that occur in single cells after the removal of Notch1 during retinal development. Retinas of Notch1fl/fl P0 pups were electroporated in vivo with plamids encoding Cre driven by a broadly active promoter, CAG, along with a Cre-responsive GFP reporter, also driven by CAG promoter (CALNL-GFP). For controls, the retinas of sibling Notch1fl/fl pups were electroporated with CAG:GFP. The animals were sacrificed at P3 to allow time for Notch1 to be genetically removed and downstream gene expression changes to occur. Retinas electroporated with either CAG;Cre and CALNL-GFP or CAG:GFP alone were dissected and dissociated to individual cells, which were harvested under a dissecting microscope on the basis of their GFP signal.

ORGANISM(S): Mus musculus

SUBMITTER: Karolina Mizeracka

PROVIDER: E-GEOD-35682 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Notch1 is required in newly postmitotic cells to inhibit the rod photoreceptor fate.

Mizeracka Karolina K DeMaso Christina R CR Cepko Constance L CL

Development (Cambridge, England) 20130703 15

Several models of cell fate determination can be invoked to explain how single retinal progenitor cells (RPCs) produce different cell types in a terminal division. To gain insight into this process, the effects of the removal of a cell fate regulator, Notch1, were studied in newly postmitotic cells using a conditional allele of Notch1 (N1-CKO) in mice. Almost all newly postmitotic N1-CKO cells became rod photoreceptors, whereas wild-type (WT) cells achieved a variety of fates. Single cell profil ...[more]

PMID: 23824579

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Dataset Information

Analysis of gene expression in wildtype and Notch1 mutant retinal cells by single cell profiling

Publications

Notch1 is required in newly postmitotic cells to inhibit the rod photoreceptor fate.

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