May 2007
Volume 48, Issue 13
Free
ARVO Annual Meeting Abstract  |   May 2007
Complement Factor H Expression Is Induced by Oxidative Stress in ARPE-19 Cells
Author Affiliations & Notes
  • M. Chrenek
    Ophthalmology, Emory University, Atlanta, Georgia
  • A. C. Ziesel
    Ophthalmology, Emory University, Atlanta, Georgia
  • J. R. Dunlevy
    Anatomy and Cell Biology, University of North Dakota, Grand Forks, North Dakota
  • P. Wong
    Ophthalmology, Emory University, Atlanta, Georgia
  • Footnotes
    Commercial Relationships M. Chrenek, None; A.C. Ziesel, None; J.R. Dunlevy, None; P. Wong, None.
  • Footnotes
    Support NSERC, FFB, RPB, Emory Eye Center, Knights Templar Foundation of Georgia, NIH P30EY-06360, UND SMHS
Investigative Ophthalmology & Visual Science May 2007, Vol.48, 5053. doi:
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      M. Chrenek, A. C. Ziesel, J. R. Dunlevy, P. Wong; Complement Factor H Expression Is Induced by Oxidative Stress in ARPE-19 Cells. Invest. Ophthalmol. Vis. Sci. 2007;48(13):5053.

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

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Abstract

Purpose:: Although Complement factor H (CFH) has been shown to have a role in age-related macular degeneration (AMD), the mechanisms of normal and aberrant CFH action are not well understood. In the current study we asked whether oxidative stress in cultured retinal pigment epithelial (RPE) cells can induce a change in expression of CFH, verified the reported expression of CFH in human retina and RPE cells by Mandal and Ayyagari (2006), and report that CFH is specifically expressed in the fovea/macular region.

Methods:: To define the spatial expression of CFH in the retina, we used quantitative RT-PCR (qRT-PCR) to measure the abundance of CFH transcript in RNA isolated from peripheral retina, fovea and RPE from human eyes. For characterization of RPE cell expression of CFH in response to oxidative stress, we treated ARPE-19 cells with 500 uM tert-butylhydroperoxide (tBH) over a 10 hour time course which is known to induce oxidative stress. RNA was extracted from the cells and qRT-PCR used to measure transcript abundance. Protein has also been extracted from the ARPE-19 cells and we plan to measure CFH protein abundance in the same profile by western blot.

Results:: We found a much higher abundance of CFH transcript in RPE as compared to retina. In ARPE-19 cells treated with tBH to induce oxidative stress, we found a 3 fold increase in CFH transcript abundance after a 10 hour treatment.

Conclusions:: CFH expression at the level of transcription is increased in response to oxidative stress in RPE cells in culture. This has important implications as to how AMD could be progressing with age. With age RPE cells that have increased levels of stress, such as the RPE cells underlying the highly active macula, may express CFH at higher levels and thereby increase the risk of developing AMD. CFH variants that have abnormal activity such as the Y402H variant may have an even greater effect.

Keywords: gene/expression • stress response • retinal degenerations: cell biology 
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