July 1996
Volume 37, Issue 8
Free
Articles  |   July 1996
The Fas-Fas ligand system and other modulators of apoptosis in the cornea.
Author Affiliations
  • S E Wilson
    Eye Institute, Cleveland Clinic Foundation, Ohio 44195, USA.
  • Q Li
    Eye Institute, Cleveland Clinic Foundation, Ohio 44195, USA.
  • J Weng
    Eye Institute, Cleveland Clinic Foundation, Ohio 44195, USA.
  • P A Barry-Lane
    Eye Institute, Cleveland Clinic Foundation, Ohio 44195, USA.
  • J V Jester
    Eye Institute, Cleveland Clinic Foundation, Ohio 44195, USA.
  • Q Liang
    Eye Institute, Cleveland Clinic Foundation, Ohio 44195, USA.
  • R J Wordinger
    Eye Institute, Cleveland Clinic Foundation, Ohio 44195, USA.
Investigative Ophthalmology & Visual Science July 1996, Vol.37, 1582-1592. doi:
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      S E Wilson, Q Li, J Weng, P A Barry-Lane, J V Jester, Q Liang, R J Wordinger; The Fas-Fas ligand system and other modulators of apoptosis in the cornea.. Invest. Ophthalmol. Vis. Sci. 1996;37(8):1582-1592.

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

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Abstract

PURPOSE: Previous studies have suggested that the disappearance of anterior keratocytes after injury to the overlying epithelium is mediated by apoptosis. The authors examined the expression of the apoptosis-related modulators, Fas (receptor), Fas ligand, Bax, Bcl-2, Bcl-XL, and interleukin-1 beta converting enzyme (ICE) in corneal cells as candidate mediators of this response and tested the effect of Fas receptor-stimulating antibody on corneal stromal fibroblast cells in vitro. METHODS: Reverse-transcription-polymerase chain reaction was used to detect FAS, FAS ligand, Bax, Bcl-2, Bcl-XL, and ICE mRNA expression in primary cultures of human corneal epithelial, stromal fibroblast, and endothelial cells. Immunohistochemistry was applied to detect Fas and Fas ligand proteins in fresh-frozen sections of normal human cornea. The effect of FAS-stimulating monoclonal antibody on first-passage stromal fibroblasts was studied using a DNA fragmentation assay, the live-dead assay with fluorescent microscopy, toluidene blue staining with light microscopy, and electron microscopy. RESULTS: FAS, Fas ligand, Bax, Bcl-2, Bcl-XL, and ICE mRNAs are expressed in all three major cell types of the cornea. Fas protein is expressed in corneal epithelial, keratocyte, and endothelial cells in fresh-frozen human cornea. Fas ligand protein, however, was detected in corneal epithelial and endothelial, but not keratocyte, cells. Fas-stimulating antibody induced first-passage stromal fibroblast cell death with morphologic changes and DNA fragmentation consistent with apoptosis. CONCLUSIONS: The Fas system (Fas and Fas ligand) modulators and final common pathway mediators of apoptosis are expressed in corneal cells. The distribution of Fas (epithelial, keratocyte, and endothelial cells) and Fas ligand (epithelial and endothelial cells) protein expression in fresh-frozen corneal tissue suggests that Fas ligand expressed in corneal epithelial and endothelial cells modulates functions in keratocyte cells and, possibly, autocrine-juxtacrine functions in epithelium and endothelium. The Fas-Fas ligand system is expressed in the cornea and could have important functions in normal corneal physiology and in the pathophysiology of corneal disease, including modulation of keratocyte apoptosis after epithelial injury.

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