May 2008
Volume 49, Issue 13
Free
ARVO Annual Meeting Abstract  |   May 2008
Wnt and Beta-Catenin Expression in the Human Limbus
Author Affiliations & Notes
  • S. L. Watson
    Department of Ophthalmology, Prince of Wales Hospital, Randwick, Australia
  • S. Sandhu
    Histology Unit, School of Medical Sciences, University of New South Wales, Randwick, Australia
  • N. Wanandy
    Histology Unit, School of Medical Sciences, University of New South Wales, Randwick, Australia
  • M. Sarris
    Histology Unit, School of Medical Sciences, University of New South Wales, Randwick, Australia
  • Footnotes
    Commercial Relationships  S.L. Watson, None; S. Sandhu, None; N. Wanandy, None; M. Sarris, None.
  • Footnotes
    Support  NHMRC Health Professional Training Fellowship, Goldstar Award University of New South Wales
Investigative Ophthalmology & Visual Science May 2008, Vol.49, 6076. doi:https://doi.org/
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    • Get Citation

      S. L. Watson, S. Sandhu, N. Wanandy, M. Sarris; Wnt and Beta-Catenin Expression in the Human Limbus. Invest. Ophthalmol. Vis. Sci. 2008;49(13):6076. doi: https://doi.org/.

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

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Abstract

Purpose: : Wnt proteins are signalling molecules that play a role in stem cell communication, corneal epithelial cell proliferation, epithelial-mesenchymal transition and ocular development. Nuclear beta-catenin is a product of Wnt signalling. Wnt4 has been found in human limbus. This study sought to confirm the presence of Wnt4 and investigate whether Wnt3, Wnt5a and beta-catenin are present in the normal adult limbus and cultured limbal epithelial cells.

Methods: : Normal human adult corneolimbal rims (n=15) were obtained. Immunohistochemistry was performed on formalin fixed, paraffin embedded tissue using the Bond-XTM automated immunostainer and Polymer Refine Detection System (Vision BioSystems). Sections were incubated with Wnt3, Wnt4, Wnt5a and beta-catenin antibodies and diaminobenzidine was used as the chromogen. Primers for Wnt3, Wnt4, and Wnt5a were obtained from Sigma - Aldrich. RT-PCR was performed on serial passages of human adult limbal epithelial cells (HLEC) with standard reagents. Slow-cycling (pulsed bromodeoxyuridine label-retaining) limbal epithelium in corneal organ culture was studied for the expression of Wnt3, Wnt4, Wnt5a and beta-catenin.

Results: : Positive immunostaining of the cytoplasm and nucleus were observed for Wnt4 in all layers of the limbal epithelium. Beta-catenin was present in the cell membranes in all layers of the corneal limbus as well as in the central corneal epithelium. Nuclear beta-catenin and Wnt5a staining were confined to a small population of cells in the basal limbus. Wnt3, Wnt4 and Wnt5a were expressed in serial passages of HLEC. Wnt4 and β-catenin were co-localised to BrdU label-retaining cells.

Keywords: cornea: epithelium • cornea: basic science • cell-cell communication 
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