June 2023
Volume 64, Issue 8
Open Access
ARVO Annual Meeting Abstract  |   June 2023
Oxidation of cysteine residues and the regulation of antioxidant defense system in RPE cells
Author Affiliations & Notes
  • sanghamitra bhattacharyya
    Cole Eye, Cleveland Clinic, Cleveland, Ohio, United States
  • Johnathon Sturgis
    Cole Eye, Cleveland Clinic Lerner College of Medicine of Case Western Reserve University, Cleveland, Ohio, United States
  • Vera L Bonilha
    Cole Eye, Cleveland Clinic Lerner College of Medicine of Case Western Reserve University, Cleveland, Ohio, United States
  • Footnotes
    Commercial Relationships   sanghamitra bhattacharyya None; Johnathon Sturgis None; Vera Bonilha None
  • Footnotes
    Support  P30EY025585(BA-A), Research to Prevent Blindness (RPB) Challenge Grant, Cleveland Eye Bank Foundation Grant, Cleveland Clinic Foundation
Investigative Ophthalmology & Visual Science June 2023, Vol.64, 2970. doi:
  • Views
  • Share
  • Tools
    • Alerts
      ×
      This feature is available to authenticated users only.
      Sign In or Create an Account ×
    • Get Citation

      sanghamitra bhattacharyya, Johnathon Sturgis, Vera L Bonilha; Oxidation of cysteine residues and the regulation of antioxidant defense system in RPE cells. Invest. Ophthalmol. Vis. Sci. 2023;64(8):2970.

      Download citation file:


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

      ×
  • Supplements
Abstract

Purpose : The retina and RPE cells are tissues with high metabolic demands constantly exposed to oxidative stress. DJ-1 is an antioxidant protein coded by the PARK7 gene. The DJ-1 cysteine (C) residues and their subsequent oxidation has been known to affect DJ-1’s cytoprotective function. However, to date, DJ-1-mediated regulation of antioxidant function in RPE cells has not been addressed substantially. Therefore, here we analyzed the impact of DJ-1 expression level and oxidation of its cysteine residues on the antioxidant role in the RPE tissues.

Methods : Endogenous DJ-1 levels were decreased with shPARK7 lentivirus. Alternatively, monolayers were also infected with adenoviruses carrying the full-length human DJ-1 cDNA (hDJ) and mutant constructs of DJ-1, which had all 3 cysteine residues mutated to serine (C2S) and also constructs with individual cysteine mutants (C46S, C53S, C106S). These monolayers were then assayed under baseline and low oxidative stress conditions (200µM H2O2 for 17hrs). The results were analyzed by immunofluorescence, Western blot, RT-PCR, mitochondrial membrane potential, and viability assays.

Results : Decreased endogenous DJ-1 levels resulted in increased levels of ROS. Furthermore, we observed morphological changes in the mitochondria structure of all the RPE monolayers transduced with all the DJ-1 constructs. The mitochondrial membrane potential of ARPE-19 monolayers overexpressing all DJ-1 constructs displayed a considerable decrease. Viability was also significantly decreased in ARPE-19 monolayers transduced with the C53S. Overexpression of the DJ-1 mutants affected the expression of antioxidant genes in ARPE-19 monolayers.

Conclusions : The antioxidant activity of DJ-1 relies on the the oxidation of C53 which is crucial for regulating endogenous levels of ROS and viability in RPE cells.

This abstract was presented at the 2023 ARVO Annual Meeting, held in New Orleans, LA, April 23-27, 2023.

×
×

This PDF is available to Subscribers Only

Sign in or purchase a subscription to access this content. ×

You must be signed into an individual account to use this feature.

×