May 2008
Volume 49, Issue 13
Free
ARVO Annual Meeting Abstract  |   May 2008
Oxidative Injury to Blood Vessels and Glia of the Optic Nerve Head in Human Glaucoma
Author Affiliations & Notes
  • Z. Feilchenfeld
    Ophthalmology & Vision Sciences, Laboratory Medicine & Pathobiology, Ophthalmic Pathology Laboratory, University of Toronto, Toronto, Ontario, Canada
  • Y. Yucel
    Ophthalmology & Vision Sciences, Laboratory Medicine & Pathobiology, Ophthalmic Pathology Laboratory, University of Toronto, Toronto, Ontario, Canada
    Keenan Research Centre at the Li Ka Shing Knowledge Institute of St. Michael's Hospital, Toronto, Ontario, Canada
  • N. Gupta
    Ophthalmology & Vision Sciences, Laboratory Medicine & Pathobiology, Ophthalmic Pathology Laboratory, University of Toronto, Toronto, Ontario, Canada
    Glaucoma & Nerve Protection Unit,
    Keenan Research Centre at the Li Ka Shing Knowledge Institute of St. Michael's Hospital, Toronto, Ontario, Canada
  • Footnotes
    Commercial Relationships  Z. Feilchenfeld, None; Y. Yucel, None; N. Gupta, None.
  • Footnotes
    Support  Institute of Medical Science Undergraduate Research Program, University of Toronto (ZF), Merck Medical School Grant (NG), Eaton Fund (NG)
Investigative Ophthalmology & Visual Science May 2008, Vol.49, 3675. doi:https://doi.org/
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    • Get Citation

      Z. Feilchenfeld, Y. Yucel, N. Gupta; Oxidative Injury to Blood Vessels and Glia of the Optic Nerve Head in Human Glaucoma. Invest. Ophthalmol. Vis. Sci. 2008;49(13):3675. doi: https://doi.org/.

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

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Abstract

Purpose: : The purpose of this study was to determine in human primary open angle glaucoma whether oxidative injury occurs in pre-laminar optic nerve blood vessels and glial cells.

Methods: : Following IRB approval, sections from post-mortem primary open angle glaucoma eyes (n=5) with mean age of 77 ± 9 yrs (±SD) were compared to age-matched normal control eyes (n=5) with mean age 73 ± 10 yrs (Eye Bank of Canada). Immunostaining with nitrotyrosine, a footprint for peroxynitrite-mediated injury, was performed and sections were double-labelled with markers for vascular endothelial cells, perivascular smooth muscle cells, and astrocytes with CD34, smooth muscle actin (SMA), and glial fibrillary acidic protein (GFAP), respectively. Immunostaining was captured in a masked fashion using confocal microscopy, in defined regions of interest for blood vessels and glial tissue. Intensity measurements of supra-threshold area in pixels as percent of the total number of pixels were measured using Image J (NIH) and compared using two-tailed Mann-Whitney nonparametric tests between glaucoma and control groups. Co-localization coefficients with cell specific markers were determined and compared with random coefficients of correlation.

Results: : Increased nitrotyrosine immunoreactivity was observed in pre-laminar optic nerve head blood vessels of primary open angle glaucoma eyes compared to controls and this difference was statistically significant (1.35 ± 1.11 % [± SD] vs. 0.01 ± 0.01 %, P = 0.008). NT immunoreactivity was also increased in the glial tissue surrounding the pre-laminar optic nerve head in the glaucoma group and compared to controls; the difference was statistically significant (18.37 ± 12.80 % vs. 0.09 ± 0.05 %, P = 0.006). Co-localization studies demonstrated nitrotyrosine staining in vascular endothelial and smooth muscle cells, in addition to astrocytes. Correlation coefficients for CD34, SMA, and GFAP were 0.37, 0.52, and 0.64 respectively.

Conclusions: : Oxidative injury is present in pre-laminar optic nerve head in human primary open angle glaucoma. The presence of peroxynitrite-mediated oxidative injury in vascular endothelial cells and smooth muscle cells, in addition to astrocytes, implicates these tissue elements in the pathobiology of glaucoma disease.

Keywords: astrocytes: optic nerve head • oxidation/oxidative or free radical damage • blood supply 
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