May 2003
Volume 44, Issue 13
Free
ARVO Annual Meeting Abstract  |   May 2003
Molecular Responses to Oxidative and Non-Oxidative Stress in the Retinal Pigment Epithelium
Author Affiliations & Notes
  • E. Chaum
    Dept of Ophthalmology, Univ of Tennessee Health Science Center, Memphis, TN, United States
  • H. Yang
    Dept of Ophthalmology, Univ of Tennessee Health Science Center, Memphis, TN, United States
  • X. Yang
    Dept of Ophthalmology, Univ of Tennessee Health Science Center, Memphis, TN, United States
  • J.C. Lang
    Alcon Research Ltd., Ft. Worth, TX, United States
  • Footnotes
    Commercial Relationships  E. Chaum, Alcon Research Ltd. Ft. Worth, TX F; H. Yang, Alcon Research Ltd., Ft. Worth, TX F; X. Yang, Alcon Research Ltd., Ft. Worth, TX F; J.C. Lang, Alcon Research Ltd., Ft. Worth, TX E.
  • Footnotes
    Support  Alcon Research Ltd. Ft. Worth, TX; NIH EY00381; Plough Foundation, Memphis
Investigative Ophthalmology & Visual Science May 2003, Vol.44, 1626. doi:
  • Views
  • Share
  • Tools
    • Alerts
      ×
      This feature is available to Subscribers Only
      Sign In or Create an Account ×
    • Get Citation

      E. Chaum, H. Yang, X. Yang, J.C. Lang; Molecular Responses to Oxidative and Non-Oxidative Stress in the Retinal Pigment Epithelium . Invest. Ophthalmol. Vis. Sci. 2003;44(13):1626.

      Download citation file:


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

      ×
  • Supplements
Abstract

Abstract: : Purpose: The focus of these studies is to characterize the molecular responses to oxidative and non-oxidative stress in the human retinal pigment epithelium (RPE). Methods: We analyzed changes in the expression of a panel of genes following oxidative stress (OS) and heat shock stress (HS) in vitro, including: Lens epithelial-derived growth factor (LEDGF), Heat shock protein 27 (Hsp27), Alpha B-crystallin (CRYAB), and Nuclear factor kappa-B (NF-kB). Confluent cultures of explanted RPE cells were treated with either 200µM H2O2 (37°C, 1 hour) or heat shock (42°C, 20 minutes). Total RNA was isolated from replicate cultures immediately following treatment, and after 2, 4, 6, and 24 hours. RT-PCR was performed with 1 µg of total cellular RNA, followed by standard amplification using gene-specific primers. Semi-quantitative analysis of the amplification products was performed on digitized images normalized to ß-actin levels and compared to controls. Results: LEDGF transcription decreased by 50% after both stressors and remained at similarly low levels over the first six hours. After 24 hours, LEDGF transcription returned to baseline levels in cells exposed to OS but increased 3-fold in cells exposed to HS. CRYAB transcription increased by 40% over the first 2 hours after HS, but decreased 40% following OS. Both treatments resulted in an eventual decline in expression to 40% of baseline over 24 hours. Hsp27 transcription increased by 40% in the first 2 hours after treatment by both stressors. Transcription remained at a steady state and elevated level in HS treated cells, but declined to baseline levels after 24 hours following OS. NF-kB transcription increased 2-fold and 5-fold following OS and HS, respectively. Transcription remained elevated after HS but declined to baseline levels after 24 hours following oxidative stress. Conclusions: The human RPE demonstrates characteristic and quantifiable molecular responses to induced sublethal stress in vitro. These studies suggest that the degree and timing of stress-specific responses correlate with specific changes in gene expression in the RPE in response to both oxidative and non-oxidative stressors.

Keywords: retinal pigment epithelium • oxidation/oxidative or free radical damage • gene/expression 
×
×

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.

×