May 2005
Volume 46, Issue 13
Free
ARVO Annual Meeting Abstract  |   May 2005
Role of PKC–epsilon in Regulating Human Corneal Epithelial Wound Healing and Barrier Function
Author Affiliations & Notes
  • J. Yin
    Departments of Ophthalmology and Anatomy and Cell Biology, Wayne State University, Detroit, MI
  • F.–S.X. Yu
    Departments of Ophthalmology and Anatomy and Cell Biology, Wayne State University, Detroit, MI
  • Footnotes
    Commercial Relationships  J. Yin, None; F.X. Yu, None.
  • Footnotes
    Support  NIH/NEI RO1 EY10869, EY14080, Research to Prevent Blindness
Investigative Ophthalmology & Visual Science May 2005, Vol.46, 2141. doi:
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      J. Yin, F.–S.X. Yu; Role of PKC–epsilon in Regulating Human Corneal Epithelial Wound Healing and Barrier Function . Invest. Ophthalmol. Vis. Sci. 2005;46(13):2141.

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

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Abstract

Abstract: : Purpose: Our previous study suggested that protein kinase C–epsilon (PKCε) is one of the isozymes responsible for phorbol 12–myristate 13–acetate–induced human corneal epithelial cell (HCEC) activation. In this study, we sought to elucidate the role of PKCε in HCEC wound healing and barrier function. Methods: HCEC lines expressing dominant negative (dn) and constitutive active (ca) PKCε, along with control cell line transfected with pcDNA3, were generated using cDNA–mediated transfection and colony selection. The effects of these mutants on cell migration and wound healing were determined by Boyden chamber and scratch wound assays, respectively. The rate of scratch wound healing was determined by the distance between the leading edge of migrating cells and the scraped edge. Measurement of trans–epithelial resistance (TER) was performed to assess the barrier function. Tight junction was evaluated by immunohistochemistry using ZO–1 antibody. Results: Compared with the control cell line, cells expressing dn–PKCε displayed an accelerated wound closure whereas the healing of a scratch wound was greatly retarded by the expression of ca–PKCε. Cells expressing dn–PKCε also exhibited an increased migrating capability than control cells in Boyden chamber assay. A significant increase in TER was observed in dn–PKCε expressing cells; while TER of ca–PKCε expressing cells was decreased, suggesting compromised barrier function. At confluence, cells expressing dn–PKCε appeared to be larger in size, whereas cells expressing ca–PKCε were morphologically similar to control cells. The staining of ZO–1 in dn–PKCε expressing cells was more pronounced and had a sharper–staining pattern as compared with control cells and cells expressing ca–PKCε. Conclusions: PKCε may play a role in corneal physiology/pathophysiology by inducing differentiation and barrier formation and/or by modulating epithelial migration and wound healing.

Keywords: cornea: epithelium • wound healing • signal transduction 
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