May 2007
Volume 48, Issue 13
ARVO Annual Meeting Abstract  |   May 2007
BMP-4: A Mediator of RPE Cell Senescence
Author Affiliations & Notes
  • D. Zhu
    Univ of Southern California, Los Angeles, California
  • S. J. Ryan
    Univ of Southern California, Los Angeles, California
    Doheny Eye Institute, Arnold and Mabel Beckman Macular Research Center, Los Angeles, California
  • D. R. Hinton
    Univ of Southern California, Los Angeles, California
    Doheny Eye Institute, Arnold and Mabel Beckman Macular Research Center, Los Angeles, California
  • Footnotes
    Commercial Relationships D. Zhu, None; S.J. Ryan, None; D.R. Hinton, None.
  • Footnotes
    Support NIH grants EY 02061 and EY 03040
Investigative Ophthalmology & Visual Science May 2007, Vol.48, 2205. doi:
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      D. Zhu, S. J. Ryan, D. R. Hinton; BMP-4: A Mediator of RPE Cell Senescence. Invest. Ophthalmol. Vis. Sci. 2007;48(13):2205.

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

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Purpose:: Bone morphogenetic protein-4 (BMP-4) is involved in a variety of important biologic processes, including programmed cell death and cell senescence. We have shown previously that BMP-4 is highly expressed in the retinal pigment epithelium (RPE) of patients with dry age-related macular degeneration (AMD). We sought to determine the effects of BMP4 on the promotion of programmed death and/or senescence of RPE cells.

Methods:: 1). ARPE-19 and human fetal RPE (fRPE) cells were treated with tert--butylhydroperoxide (tBH, 30 uM) and hydrogen peroxide (H2O2, 150 uM) in culture medium containing 10% fetal bovine serum for 2 hours for each stress. The cells were serially stressed five times with tBH treatment and three times with H2O2 treatment. 2). BMP4 overexpressed ARPE-19 stable cell lines (ARPE-BMP4) were established by transducing BMP4-lentiviral constructs into ARPE-19 cells. BMP4-lentiviral vector was constructed by inserting BMP4 cDNA into the pLenti6/V5-TOPO vector (Invitrogen). 3). Western blot and real time RT-PCR were used to semiquantitate or quantitate the expression levels of target genes. 4). SA-ß-Gal staining assay was used to detect the senescent cells.

Results:: Western blot showed significantly increased expression and secretion of recombinant BMP4 protein in ARPE-BMP4 cells. ß-galactosidase activity was dramatically increased in ARPE-BMP4 cells when the cells were cultured in serum free ARPE medium for one week. BMP4 expression in ARPE-19 and fRPE cells increased about eight-fold after tBH or H2O2 treatment compared to that of the non-stressed cells. tBH and H2O2 induced irreversible cell growth arrest and premature senescence of ARPE-19 and fRPE cells.

Conclusions:: Both BMP4 and oxidative stress induce RPE cell senescence. Oxidative stress also increases the BMP4 expression in human RPE cells. The results indicate that the expression of RPE senescence-related genes or biomarkers under oxidative stress may be mediated by the BMP4 pathway.

Keywords: age-related macular degeneration • growth factors/growth factor receptors • retinal pigment epithelium 

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