May 2008
Volume 49, Issue 13
Free
ARVO Annual Meeting Abstract  |   May 2008
Up-Regulation of ZO-1 by the Fibronectin-Derived Peptide PHSRN in Cultured Human Corneal Epithelial Cells
Author Affiliations & Notes
  • R. Yanai
    Ophthalmology, Yamaguchi University Graduate School of Medicine, Ube City, Japan
  • J.-A. Ko
    Ophthalmology, Yamaguchi University Graduate School of Medicine, Ube City, Japan
  • T.-I. Chikama
    Ocular Pathophysiology, Yamaguchi University, Ube City, Japan
  • T. Nishida
    Ophthalmology, Yamaguchi University Graduate School of Medicine, Ube City, Japan
  • Footnotes
    Commercial Relationships  R. Yanai, None; J. Ko, None; T. Chikama, None; T. Nishida, None.
  • Footnotes
    Support  None.
Investigative Ophthalmology & Visual Science May 2008, Vol.49, 3373. doi:https://doi.org/
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      R. Yanai, J.-A. Ko, T.-I. Chikama, T. Nishida; Up-Regulation of ZO-1 by the Fibronectin-Derived Peptide PHSRN in Cultured Human Corneal Epithelial Cells. Invest. Ophthalmol. Vis. Sci. 2008;49(13):3373. doi: https://doi.org/.

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

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Abstract

Purpose: : The barrier function of epithelia depends primarily on tight junctions, of which zonula occludens (ZO)-1 is a major component. We have previously shown that the peptide PHSRN, which corresponds to the second cell-binding domain of fibronectin, stimulates both migration of the corneal epithelium in vitro and corneal epithelial wound healing in vivo. We have now investigated the effect of PHSRN on ZO-1 expression in cultured human corneal epithelial cells.

Methods: : Simian virus 40-transformed human corneal epithelial (HCE) cells were cultured with PHSRN or the control peptide NRSHP. Expression of ZO-1 was determined at the mRNA and protein levels by reverse transcription-polymerase chain reaction analysis and by both immunoblot analysis and flow cytometry, respectively. Phosphorylation of the signaling proteins ERK, JNK, p38 MAPK, and c-Jun was measured with the Bio-Plex system (Bio-Rad) and by immunoblot analysis.

Results: : Exposure of HCE cells to PHSRN increased the abundance of ZO-1 mRNA in a concentration- and time-dependent manner. Incubation of the cells with the control peptide NRSHP had no effect on the amount of ZO-1 mRNA. PHSRN, but not NRSHP, also increased the abundance of ZO-1 protein in a concentration- and time-dependent manner. Exposure of the cells to PHSRN increased the phosphorylation of ERK, JNK, p38 MAPK, and c-Jun.

Conclusions: : The fibronectin-derived peptide PHSRN up-regulates the expression of ZO-1 at the protein and mRNA levels in cultured human corneal epithelial cells. The effect of PHSRN on ZO-1 expression was accompanied by activation of mitogen-activated protein kinase cascades and the transcription factor c-Jun. In addition to its stimulation of the migration of corneal epithelial cells, PHSRN may thus also promote the formation of tight junctions between these cells.

Keywords: cornea: epithelium • wound healing • cell adhesions/cell junctions 
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