R-spondin 2 is required for optic vesicle morphogenesis and neuroretina specification in vivo and in vitro

  • Deepti Abbey (Contributor)
  • Geoffrey A. Neale (Contributor)
  • Guillermo Oliver (Contributor)



Accession Number: GSE97209

GPL16570: [MoGene-2_0-st] Affymetrix Mouse Gene 2.0 ST Array [transcript (gene) version]

Organism: Mus musculus

Published on 2017-11-07

Using stem cell–based therapies to treat retinal abnormalities is becoming a likely possibility; therefore, identifying the key factors and the relevant mechanisms controlling optic vesicle morphogenesis and neuroretina (NR) differentiation is important. Recent advances in self-organizing, 3-dimensional (3D) tissue cultures of embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs) provided a valuable in vitro model for characterizing regulatory cascades and signaling pathways controlling mammalian retinal development. Using Rx-GFP expressing ESCs and Six3–/– iPSCs we identified R-spondin 2 (Rspo2)-mediated repression of Wnt signaling as a novel required step during optic vesicle morphogenesis and NR differentiation. Furthermore, we also show that transient ectopic expression of Rspo2 in the anterior neural plate of transgenic mouse embryos was sufficient to arrest NR differentiation. ChIP assays identified Six3-responsive elements in the Rspo2-promoter region, indicating that Six3-mediated repression of Rspo2 is required to restrict Wnt signaling in the developing anterior neuroectoderm and allow eye development to proceed.

Overall Design:
We used microarrays to measure changes in gene expression profiles in mouse embryonic stem cells after retinal differentiation for 4 or 7 days. Mouse ES cells were derived from the mouse embryonic stem cell line EB5 (129/Ola) in which GFP gene is knocked-in under Rx promoter (Eiraku and Sasai, Nat Protocols 7:69-79, 2012; RIKEN cell line AES0145). Expression profiles were compared at day 4 and day 7 of retinal differentiation between WT Rx-GFP ESC.

Name: Geoffrey Neale
Organization: St Jude Childrens Research Hospital
Deparment: Hartwell Center
Address: 262 Danny Thomas Place Memphis TN 38107 USA
Email: geoffrey.neale@stjude.org

Organization: Affymetrix, Inc.
Address: Santa Clara CA 95051 USA
Email: geo@ncbi.nlm.nih.gov, support@affymetrix.com
Phone: 888-362-2447
Web-Link: http://www.affymetrix.com/index.affx
Date made availableMar 29 2017
PublisherGene Expression Omnibus

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