April 2011
Volume 52, Issue 14
Free
ARVO Annual Meeting Abstract  |   April 2011
Characterization of the DJ-1 Function in the Retina and Retinal Pigment Epithelium
Author Affiliations & Notes
  • Vera L. Bonilha
    Ophthalmology, Cole Eye Inst/Cleveland Clin Lerner Ctr, Cleveland, Ohio
  • Mary E. Rayborn
    Ophthalmology, Cole Eye Inst/Cleveland Clin Lerner Ctr, Cleveland, Ohio
  • Yong Li
    Ophthalmology, Cole Eye Inst/Cleveland Clin Lerner Ctr, Cleveland, Ohio
  • Chengsong Xie
    Lab. of Neurogenetics, National Institute on Aging, NIH, Bethesda, Maryland
  • Huaibin Cai
    Lab. of Neurogenetics, National Institute on Aging, NIH, Bethesda, Maryland
  • Footnotes
    Commercial Relationships  Vera L. Bonilha, None; Mary E. Rayborn, None; Yong Li, None; Chengsong Xie, None; Huaibin Cai, None
  • Footnotes
    Support  Supported in part by NIH grants EY017153 and EY15638, a Research Center Grant from The Foundation Fighting Blindness; a Research to Prevent Blindness Unrestricted Grant and funds from the Cleveland Cl
Investigative Ophthalmology & Visual Science April 2011, Vol.52, 4781. doi:
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      Vera L. Bonilha, Mary E. Rayborn, Yong Li, Chengsong Xie, Huaibin Cai; Characterization of the DJ-1 Function in the Retina and Retinal Pigment Epithelium. Invest. Ophthalmol. Vis. Sci. 2011;52(14):4781.

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

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Abstract

Purpose: : DJ-1 was first discovered as a novel oncogene product that transforms mouse NIH3T3 cells in cooperation with activated ras and was later identified as a causative gene of familial Parkinson’s disease. DJ-1 is a multifunctional protein, which has been shown to robustly protect cells from oxidative stress in several cell types. DJ-1 is ubiquitously expressed in several tissues, with high expression in the testis and astrocytes of the frontal cortex. DJ-1 peptides were detected in rat retinal pigment epithelium (RPE) cell fractions subjected to proteomic analysis. The present study was conducted to further analyze the localization and function of DJ-1 in mouse retinal tissues and RPE cells in culture.

Methods: : Cryosections of control and DJ-1 knockout (KO) mouse retinas were processed for immunofluorescence with DJ-1 specific antibodies and other cell markers followed by confocal microscopy analysis. The presence of DJ-1 was analyzed in whole cell lysates from retina, RPE, and several RPE cell lines. To decipher the DJ-1 function, RPE cultures were treated with H2O2 (100 to 800uM) and 4-HNE (5 to 100uM) for various times followed by biochemical and immunohistological analysis. In addition, cells were infected with a replication-deficient adenovirus carrying the human DJ-1 cDNA prior to stress experiments. Results were analyzed by immunofluorescence, and Westerns.

Results: : DJ-1 immunoreactivity was observed with cells in the ganglion cell layer, cells in the inner nuclear layer, photoreceptors and RPE. In RPE cells under baseline conditions, DJ-1 displays a diffuse cytoplasmic and nuclear staining. Upon oxidative injury, a large portion of DJ-1 redistributed to mitochondria. A major band of ~ 25kDa was observed in the extracts of mouse retina and RPE and in all the RPE cell lines as compared to extracts from mouse brain. Increase in DJ-1 expression was observed when cells were exposed to oxidative stress as analyzed by immunocytochemical and biochemical assays. Overexpression of DJ-1 prior to exposure to oxidative stress led to significant decrease in the generation of reactive oxidative stress species and oxidative stress-related cellular damage. Finally, histological evaluation of the retina in young DJ-1 KO revealed vesicle accumulation and shrinking of outer plexiform layer; RPE pyknosis and photoreceptor cell nuclei death.

Conclusions: : DJ-1 expression is increased upon exposure to oxidative stress. In addition, DJ-1 expression promotes protection of RPE cells to oxidative stress.

Keywords: oxidation/oxidative or free radical damage • protein structure/function • retinal pigment epithelium 
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