May 2004
Volume 45, Issue 13
Free
ARVO Annual Meeting Abstract  |   May 2004
Serum Deprivation Induced Apoptosis Signaling Pathways of RGC–5 Retinal Ganglion Cells.
Author Affiliations & Notes
  • N. Agarwal
    Cell Biology & Genetics,
    UNT Health Science Center, Fort Worth, TX
  • I. Charles
    Cell Biology & Genetics,
    UNT Health Science Center, Fort Worth, TX
  • D.M. Kumar
    Cell Biology & Genetics,
    UNT Health Science Center, Fort Worth, TX
  • R.R. Krishnamoorthy
    Pharmacology & Neuroscience,
    UNT Health Science Center, Fort Worth, TX
  • R.S. Roque
    Cell Biology & Genetics,
    UNT Health Science Center, Fort Worth, TX
  • Footnotes
    Commercial Relationships  N. Agarwal, None; I. Charles, None; D.M. Kumar, None; R.R. Krishnamoorthy, None; R.S. Roque, None.
  • Footnotes
    Support  American Health Assistance Foundation–National Glaucoma Program
Investigative Ophthalmology & Visual Science May 2004, Vol.45, 2165. doi:
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    • Get Citation

      N. Agarwal, I. Charles, D.M. Kumar, R.R. Krishnamoorthy, R.S. Roque; Serum Deprivation Induced Apoptosis Signaling Pathways of RGC–5 Retinal Ganglion Cells. . Invest. Ophthalmol. Vis. Sci. 2004;45(13):2165.

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

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Abstract

Abstract: : Purpose: To determine the signaling pathways of apoptosis in retinal ganglion cells deprived of growth factors. Methods: An established line of transformed rat retinal ganglion cells, RGC–5, was subjected to serum deprivation for 3–6 days. Control cells were cultured in growth medium containing 10% fetal calf serum. Cell viability was measured by neutral red assay, apoptotic signaling by immunoblot analyses, electrophoretic mobility shift assay, and biochemical methods. Results: Serum deprivation of RGC–5 cells for three days resulted in 50% cell loss due to apoptosis as established by DNA laddering and propidium iodide staining. Serum deprivation also resulted in increased oxidative stress as revealed by lowering of reduced glutathione (GSH) and increased levels of malonydialdehyde (MDA). Reduced levels of NF–kB binding activity was observed upon serum deprivation of RGC–5 cells. Serum deprivation was also associated with a loss of mitochondrial function as revealed by cytochrome–c release and rhodamine 123 staining. The RGC–5 cell death was further augmented by PI3K inhibitor (LY 294002), MAPK inhibitor (PD 098059), and a Trk– receptor inhibitor (K252a). Serum deprivation also resulted in increased phosphorylation of ERK proteins. Conclusions: These results indicate that serum deprivation of RGC–5 cells result in apoptotic cell death by means of mitochondrial pathways involving oxidative stress, PI3K, MAPK and trk receptors. Supported by American Health Assistance Foundation–National Glaucoma Program (NA).

Keywords: cell death/apoptosis • signal transduction • ganglion cells 
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