April 2014
Volume 55, Issue 13
Free
ARVO Annual Meeting Abstract  |   April 2014
Modulation of the Notch and Wnt signalling by TGF- β in adult human Müller stem cells.
Author Affiliations & Notes
  • NA WU
    National Institute for Health Research (NIHR) Biomedical Research Centre at Moorfields Eye Hospital NHS Foundation Trust and UCL Institute of Ophthalmology, UCL, London, United Kingdom
    Department of Ophthalmology and Visual Science, Eye & ENT Hospital, Shanghai Medical College, Fudan University, Fudan University, Shanghai, China
  • Joseph Wiseman
    National Institute for Health Research (NIHR) Biomedical Research Centre at Moorfields Eye Hospital NHS Foundation Trust and UCL Institute of Ophthalmology, UCL, London, United Kingdom
  • Yuan Lei
    Department of Ophthalmology and Visual Science, Eye & ENT Hospital, Shanghai Medical College, Fudan University, Fudan University, Shanghai, China
  • Karen Eastlake
    National Institute for Health Research (NIHR) Biomedical Research Centre at Moorfields Eye Hospital NHS Foundation Trust and UCL Institute of Ophthalmology, UCL, London, United Kingdom
  • Xinghuai Sun
    Department of Ophthalmology and Visual Science, Eye & ENT Hospital, Shanghai Medical College, Fudan University, Fudan University, Shanghai, China
  • G Astrid Limb
    National Institute for Health Research (NIHR) Biomedical Research Centre at Moorfields Eye Hospital NHS Foundation Trust and UCL Institute of Ophthalmology, UCL, London, United Kingdom
  • Footnotes
    Commercial Relationships NA WU, None; Joseph Wiseman, None; Yuan Lei, None; Karen Eastlake, None; Xinghuai Sun, None; G Astrid Limb, None
  • Footnotes
    Support None
Investigative Ophthalmology & Visual Science April 2014, Vol.55, 1373. doi:
  • Views
  • Share
  • Tools
    • Alerts
      ×
      This feature is available to authenticated users only.
      Sign In or Create an Account ×
    • Get Citation

      NA WU, Joseph Wiseman, Yuan Lei, Karen Eastlake, Xinghuai Sun, G Astrid Limb; Modulation of the Notch and Wnt signalling by TGF- β in adult human Müller stem cells.. Invest. Ophthalmol. Vis. Sci. 2014;55(13):1373.

      Download citation file:


      © ARVO (1962-2015); The Authors (2016-present)

      ×
  • Supplements
Abstract

Purpose: The transforming growth factor β (TGF- β) and the Wnt, Notch signalling pathways all play crucial roles in many biological processes, including embryonic development, cell fate differentiation and cell proliferation. There is evidence for a crosstalk between the Notch and Wnt pathways during retinal development, but it is not known whether co-activation of these pathways may occur in adult human Müller stem cells (hMSCs) or whether these can be modulated by TGF- β.It was therefore the aim of this study to investigate the effect of this cytokine on the expression of molecules involved in these two pathways

Methods: Human Müller stem cells were cultured on matrigel using DMEM containing foetal calf serum (FCS). hMSCs were cultured in the absence or presence of the γ-secretase inhibitor DAPT plus bFGF to induce their differentiation into RGCs. Exogenous TGF- β1 or TGF- β3 were added to the differentiating or control cells which were cultured for up to 7 days. mRNA and protein extracted from these cells were examined by RT-PCR, western-blot and immuno-staining techniques for the expression of molecules of the Notch and Wnt signalling pathways. The hexosaminidase assay and Ki67 immuno-staining were used to test the effect of TGF- β on cell proliferation.

Results: Notch inhibition by DAPT caused down-regulation in gene expression of Hes1 and Wnt 2b. Similarly, TGF- β1 alone caused a decrease in gene and protein expression of the Notch downstream target Hes1 and the Wnt signalling ligand Wnt2b, while inducing an increase of the canonical Wnt signalling intracellular component β-catenin. Down-regulation of these genes by DAPT was not modified by addition of TGF- β1 or TGF- β3 to the cultured cells. It was also observed that both TGF- β1 and TGF- β3 had a similar effect on the inhibition of cell growth.

Conclusions: These results provide evidence that TGF- β directly induces down-regulation of the Notch target Hes1 and the Wnt signalling ligand Wnt2b, as well as upregulation of β-catenin. Furthermore, down-regulation of Notch by DAPT results in down-regulation of Wnt2b, suggesting a crosstalk between these two signalling pathways. However, TGF- β does not have additive effects on the down-regulation of Hes1 and Wnt2b by DAPT. The observations suggest that these two molecules may activate similar intracellular pathways mediating neural differentiation of Müller stem cells.

Keywords: 721 stem cells • 603 Muller cells  
×
×

This PDF is available to Subscribers Only

Sign in or purchase a subscription to access this content. ×

You must be signed into an individual account to use this feature.

×