May 2004
Volume 45, Issue 13
Free
ARVO Annual Meeting Abstract  |   May 2004
Adenovirus–mediated delivery of catalase protects the mouse retina in vivo and RPE cells in vitro from oxidative stress induced cell death
Author Affiliations & Notes
  • I. Tsui
    F.M Kirby Center for Molecular Ophthalmology, University of Pennsylvania, Philadelphia, PA
  • T. Rex
    F.M Kirby Center for Molecular Ophthalmology, University of Pennsylvania, Philadelphia, PA
  • A. Maguire
    F.M Kirby Center for Molecular Ophthalmology, University of Pennsylvania, Philadelphia, PA
  • J. Bennett
    F.M Kirby Center for Molecular Ophthalmology, University of Pennsylvania, Philadelphia, PA
  • J.L. Dunaief
    F.M Kirby Center for Molecular Ophthalmology, University of Pennsylvania, Philadelphia, PA
  • Footnotes
    Commercial Relationships  I. Tsui, None; T. Rex, None; A. Maguire, None; J. Bennett, None; J.L. Dunaief, None.
  • Footnotes
    Support  NIH EY00417, RPB, IRRF, The Steinbach Foundation
Investigative Ophthalmology & Visual Science May 2004, Vol.45, 3699. doi:
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      I. Tsui, T. Rex, A. Maguire, J. Bennett, J.L. Dunaief; Adenovirus–mediated delivery of catalase protects the mouse retina in vivo and RPE cells in vitro from oxidative stress induced cell death . Invest. Ophthalmol. Vis. Sci. 2004;45(13):3699.

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

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Abstract

Abstract: : Purpose: To determine whether over–expression of the antioxidant enzyme catalase protects the retina from oxidative stress Methods:(in vivo) Ad.CMV.catalase or Ad.lacZ were injected into the subretinal space of Balb/c mice 1 day prior to light damage. Mice were dark adapted and then subjected to 7 hours of 8,000 lux white fluorescent light. 24 hours after light damage, a group of eyes was fixed in 4% paraformaldehyde and stained with TUNEL and anti–catalase. Seven days after light damage, another group of eyes was fixed in 2% glutaraldehyde/2% paraformaldehyde, embedded in paraffin, and stained with hematoxylin and eosin. (in vitro) ARPE19 cells were plated on chamber slides and exposed to Ad.CMV.catalase, Ad.GFP, or no virus. 24 hours after infection, the cells were subjected to 1 mM H2O2 for 12 hours. Cells were fixed in 2% gluteraldehyde/2% paraformaldehyde and stained with TUNEL and anti–catalase. Results:At 24 hours after light damage, many TUNEL–positive nuclei were found in light–damaged retinas injected with Ad.lacZ, but very few were found in light–damaged retinas injected with Ad.CMV.catalase. At 7 days after light damage, eyes injected with Ad.lacZ showed loss of inner and outer segments and thinning of the outer nuclear layer. In contrast, eyes injected with Ad.CMV.catalase had retinas with normal architecture. In vitro, there were fewer TUNEL positive cells surrounding RPE cells which over –expressed catalase. Conclusions:In vivo, catalase over–expression in the RPE by gene transfer with Ad.CMV.catalase is able to prevent light damage induced photoreceptor death. In vitro, catalase over–expressing RPE cells can protect their neighbors, possibly through diffusion of H2O2 down a concentration gradient. These results implicate hydrogen peroxide, the catalase substrate, in the pathogenesis of photic injury and provide a paradigm for anti–oxidative gene therapy in retinal degenerations involving oxidative stress.

Keywords: adenovirus • antioxidants • photoreceptors 
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