Oxidative injury to blood vessels and glia of the pre-laminar optic nerve head in human glaucoma

Experimental Eye Research
Zac FeilchenfeldNeeru Gupta

Abstract

Glaucoma is a leading cause of irreversible world blindness. Oxidative damage and vascular injury have been implicated in the pathogenesis of this disease. 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. 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 normal control eyes (n=4) with mean age 70 +/- 9 yrs (Eye Bank of Canada). Immunostaining with nitrotyrosine, a footprint for peroxynitrite-mediated injury, was performed and sections were double-labeled 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, and 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 calculated using ImageJ (NIH) and compared using two-tailed Mann-Whitney nonparametric tests between glaucoma and control groups. Colocal...Continue Reading

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