June 2013
Volume 54, Issue 15
Free
ARVO Annual Meeting Abstract  |   June 2013
Localisation of Yap/Taz in corneal epithelia: a marker of mechano-sensitivity and role in epithelial homeostasis
Author Affiliations & Notes
  • Che Connon
    School of Chemistry, Food and Pharmacy, University of Reading, Reading, United Kingdom
  • Roanne Jones
    School of Chemistry, Food and Pharmacy, University of Reading, Reading, United Kingdom
  • James Foster
    School of Chemistry, Food and Pharmacy, University of Reading, Reading, United Kingdom
  • Footnotes
    Commercial Relationships Che Connon, None; Roanne Jones, None; James Foster, None
  • Footnotes
    Support None
Investigative Ophthalmology & Visual Science June 2013, Vol.54, 2582. doi:
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      Che Connon, Roanne Jones, James Foster, ; Localisation of Yap/Taz in corneal epithelia: a marker of mechano-sensitivity and role in epithelial homeostasis. Invest. Ophthalmol. Vis. Sci. 2013;54(15):2582.

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

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Abstract

Purpose: To investigate the cellular location of the nuclear transcription factor Yap/Taz in limbal and central corneal epithelial cells and relate this expression to changes in corneal stiffness centripetally across the ocular surface. Yap/Taz has recently been found to be an important regulator of mechanotransduction. We believe that changes in substrate stiffness occur across the cornea and that a centripetal stiffness gradient drives both differentiation and migration of epithelial cells form the limbus towards the central cornea forming an important structural component of ocular surface homeostasis.

Methods: The localisation of Yap/Taz, CK3, CK14 and ZO-1 across the ocular surface of bovine corneas was examined by immunohistochemistry. Limbal stem cells were isolated form fresh bovine corneas and expanded upon type I collagen gels of differing stiffness for 14 days. The localisation of Yap/Taz, CK3, CK14 and ZO-1 was examined by immunohistochemistry within these corneal constructs. Furthermore, the transcription levels of Yap/Taz, CK3 and ABCG2 were quantified by QPCR.

Results: Across the healthy bovine cornea Yap/Taz was predominately expressed cytoplasmically within the limbus, where as in central corneal epithelial cells Yap/Taz was retained within the nucleus. Isolated limbal epithelial cells expanded upon the more compliant collagen gels showed significantly less gene expression of Yap/Taz, which was predominately cytoplasimic at the protein level, whereas more nuclear expression was seen within epithelial cells expanded upon the stiffer collagen gels. This corresponded with more cells expressing cytokeratin 3 and ZO-1 and less cytokeratin 14 and ABCG2 at the gene and protein level.

Conclusions: The nuclear to cytoplasmic expression ratio of Yap/Taz between limbal and central epithelial cells supports the notion of a centripetal stiffness gradient across the corneal surface. The suitability of YAP/Taz as a marker of mechanosensitivity (substrate stiffness) in corneal epithelial cells was successfully demonstrated by use of collagen gels as cell substrates with differing stiffness. The presence of a centripetal stiffness gradient across the cornea is likely to underpin new directions in corneal wound healing and our understanding of ocular surface homeostasis.

Keywords: 480 cornea: basic science • 482 cornea: epithelium • 500 differentiation  
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