May 2005
Volume 46, Issue 13
Free
ARVO Annual Meeting Abstract  |   May 2005
Oxidative Stress Triggers the Expression of Bcl–2 Family Proteins in Retinal Pigment Epithelial Cells (RPE)
Author Affiliations & Notes
  • R. Antony
    Neuroscience, LSU Health Sciences Center – New Orleans, New Orleans, LA
  • M. Hardy
    Neuroscience, LSU Health Sciences Center – New Orleans, New Orleans, LA
  • N.G. Bazan
    Neuroscience, LSU Health Sciences Center – New Orleans, New Orleans, LA
  • Footnotes
    Commercial Relationships  R. Antony, None; M. Hardy, None; N.G. Bazan, None.
  • Footnotes
    Support  NIH grant EY05121
Investigative Ophthalmology & Visual Science May 2005, Vol.46, 5357. doi:
  • Views
  • Share
  • Tools
    • Alerts
      ×
      This feature is available to authenticated users only.
      Sign In or Create an Account ×
    • Get Citation

      R. Antony, M. Hardy, N.G. Bazan; Oxidative Stress Triggers the Expression of Bcl–2 Family Proteins in Retinal Pigment Epithelial Cells (RPE) . Invest. Ophthalmol. Vis. Sci. 2005;46(13):5357.

      Download citation file:


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

      ×
  • Supplements
Abstract

Abstract: : Purpose: Previous works from our lab have shown that apoptosis is triggered in RPE cells when exposed to oxidative stress (PNAS, 101:8491–8496, 2004). Bcl–2 family proteins are expressed when cells are exposed to a variety of stimuli. Some of these proteins are anti–apoptotic and others are pro–apoptotic. Our goal in this study was to investigate which members of the Bcl–2 family proteins are triggered by oxidative stress and TNF–α in ARPE–19 cells. We also aimed at defining cellular localization of the Bcl–2 protein by immunohistochemistry. Methods: Human RPE cells were grown in a 37 °C humidified chamber with 5% CO2 in Dulbecco’s modified Eagle’s medium (DMEM). Cells were grown on chamber glass slides (LAB–TEK) with 80% confluence. Cells were treated with TNF–α (10 µg/ml) and H2O2 (0.8 mM); untreated cells were controls. The incubation lasted for 16 hours. Immunostaining was done by using primary antibodies against Bcl–2, Bax, Bcl–xL, Bad (Santa Cruz), and Cytochrome C proteins at 1:100 dilution for 2 hours. Some of the antibodies were conjugated to FITC or TRITC. Cells were viewed using a Zeiss deconvolution microscope. Results: TNF–α / H2O2 triggers the induction of the Bcl–2 family proteins. Our study demonstrates that RPE cells under oxidative stress show a significant increase (10 fold) in the anti–apoptotic proteins Bcl–xL and Bcl–2. Pro–apoptotic proteins Bad and Bax expression levels also increased (at least 20 fold) with cytoplasmic condensation and nuclear shrinkage. Conclusions: Oxidative stress in RPE cells significantly increased the expression of anti–apoptotic members of the Bcl–2 family of proteins. The induction of anti–apoptotic members suggests a protective attempt to counteract the effects of oxidative stress from homeostasis. Supported by NIH EY05121

Keywords: retinal pigment epithelium • oxidation/oxidative or free radical damage • apoptosis/cell death 
×
×

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.

×