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J.M. Barnett, J.D. Kay, V.S. Rajaratnam, K.A. Koepke, J.S. Penn; The Influence of Penetrating Ocular Injury on the Spatial and Temporal Distribution of PEDF mRNA and Protein in the Oxygen Injured Rat Retina . Invest. Ophthalmol. Vis. Sci. 2004;45(13):1888.
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© ARVO (1962-2015); The Authors (2016-present)
Purpose: Penetrating ocular injury produces an angiostatic effect potentially caused by release of endogenous antiangiogenic factors from the wound site (Penn JS, ARVO Abs 510, ). We sought to determine the spatial and temporal distribution of pigment epithelium–derived factor (PEDF) in order to determine its consistency with the pattern of this angiostatic effect. Methods: Sprague–Dawley rat litters were exposed to alternating 50% and 10% oxygen atmospheres from birth through P14. Upon removal from the exposure chamber, rats received a penetrating dry needle puncture at the temporal pole of the eye, and the rats were sacrificed 1,3, and 6 days later. Some of the eyes were pooled for analysis of retinal RNA by Northern blotting, and retinas and vitreous samples from other eyes were pooled for Western blotting. Still other eyes were sectioned for analysis by in situ hybridization to determine the spatial distribution of PEDF mRNA relative to the wound site. Results: Needle puncture caused a 4.1–fold increase in retinal PEDF mRNA and a 2.1–fold increase in retinal PEDF protein 1 day after injury. Vitreous PEDF protein was increased by 3.4X in punctured eyes vs. controls at this time. At day 1 post–injury, in situ hybridization showed an increase in PEDF mRNA in areas surrounding the puncture site. Expression was increased in both the RPE and ganglion cell layers; this increase was not seen in uninjured areas or in unpunctured eyes. Conclusions: In situ hybridization and Northern and Western blots revealed temporal and spatial distributions of retinal PEDF mRNA and protein consistent with the angiostatic effect of penetrating ocular injury. Further study may reveal how PEDF induction participates in penetrating ocular injury and may provide additional insight into its physiologic roles.
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