December 2002
Volume 43, Issue 13
ARVO Annual Meeting Abstract  |   December 2002
Increased Apoptosis in Retinal Pigment Epithelial Cells With Reduced Antioxidant Enzymes
Author Affiliations & Notes
  • WF Obritsch
    Ophthalmology University of Minnesota Minneapolis MN
  • SJ Coloso
    Ophthalmology University of Minnesota Minneapolis MN
  • H Van Remmen
    Grecc South Texas Veterans Health Care System San Antonio TX
  • J Yang
    Ophthalmology University of Minnesota Minneapolis MN
  • DS Gregerson
    Ophthalmology University of Minnesota Minneapolis MN
  • DA Ferrington
    Ophthalmology University of Minnesota Minneapolis MN
  • Footnotes
    Commercial Relationships   W.F. Obritsch, None; S.J. Coloso, None; H. Van Remmen, None; J. Yang, None; D.S. Gregerson, None; D.A. Ferrington, None. Grant Identification: Minnesota Medical Foundation and Research to Prevent Blindness
Investigative Ophthalmology & Visual Science December 2002, Vol.43, 674. doi:
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    • Get Citation

      WF Obritsch, SJ Coloso, H Van Remmen, J Yang, DS Gregerson, DA Ferrington; Increased Apoptosis in Retinal Pigment Epithelial Cells With Reduced Antioxidant Enzymes . Invest. Ophthalmol. Vis. Sci. 2002;43(13):674.

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

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Abstract: : Purpose: Apoptosis in retinal pigment epithelial (RPE) cells has been proposed to be a mechanism for age-related macular degeneration, and some supportive evidence has been reported. Oxidative stress has been shown to induce apoptosis in cultured RPE cells. We postulated that decreased protection by antioxidant enzymes could place the RPE at greater risk for apoptotic cell death. Methods: RPE cells were isolated from wild type and transgenic knock-out (KO) mice expressing decreased levels of either copper-zinc superoxide dismutase (SOD1 +/-) or glutathione peroxidase (GPX1 -/-). Cells were transformed, grown to confluence, then exposed to varying levels of hydrogen peroxide (1-4 mM) for one hour. The relative concentration of intracellular hydrogen peroxide was monitored using dichlorodihydrofluorescein (DCHF). Apoptosis was determined by quantification of DNA fragmentation by FACS analysis of TUNEL positive cells. Cell membrane permeability was evaluated from propidium iodide (PI) fluorescence. Results: Following exposure of RPE cells to increasing concentrations of hydrogen peroxide, we observed a peroxide dose-dependent response in DCHF fluorescence, confirming an intracellular increase in hydrogen peroxide. Four hours after peroxide stress, there was a two-fold increase in TUNEL positive cells for RPE from KO mice compared with RPE from wild type mice. There was a peroxide dose-dependent increase in PI fluorescence in all cell lines, with the greatest increase exhibited in RPE from GPX1 mice. Conclusion: RPE cells with decreased levels of copper-zinc SOD and GPX are more susceptible to peroxide-induced apoptotic cell death. Decreased GPX activity may place RPE cells at a greater risk for membrane damage during oxidative stress.

Keywords: 567 retinal pigment epithelium • 504 oxidation/oxidative or free radical damage • 321 antioxidants 

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