April 2011
Volume 52, Issue 14
Free
ARVO Annual Meeting Abstract  |   April 2011
Light-induced Death Of Cells In Culture Is By Necroptosis Rather Than Apoptosis
Author Affiliations & Notes
  • Neville N. Osborne
    Nuffield Lab of Ophthalmology, University of Oxford, Oxford, United Kingdom
    Fundación de Investigación Oftalmológica, Instituto Oftalmológico Fernández-Vega, Oviedo 330, Spain
  • Susana del Olmo-Aguado
    Fundación de Investigación Oftalmológica, Instituto Oftalmológico Fernández-Vega, Oviedo 330, Spain
  • Tengku A. Kamalden
    Nuffield Lab of Ophthalmology, University of Oxford, Oxford, United Kingdom
  • Dan Ji
    Nuffield Lab of Ophthalmology, University of Oxford, Oxford, United Kingdom
  • Footnotes
    Commercial Relationships  Neville N. Osborne, None; Susana del Olmo-Aguado, None; Tengku A. Kamalden, None; Dan Ji, None
  • Footnotes
    Support  NNO:Cátedra de Biomedicina (Chair of Biomedicine), supported by the Fundación BBVA, Spain.
Investigative Ophthalmology & Visual Science April 2011, Vol.52, 3084. doi:
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    • Get Citation

      Neville N. Osborne, Susana del Olmo-Aguado, Tengku A. Kamalden, Dan Ji; Light-induced Death Of Cells In Culture Is By Necroptosis Rather Than Apoptosis. Invest. Ophthalmol. Vis. Sci. 2011;52(14):3084.

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

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Abstract

Purpose: : Purpose: Characterise the mechanism by which light as impinging on the retina in situ kills RGC-5 cells (cell line with certain ganglion cell properties) in vitro.

Methods: : Methods: RGC-5 cell cultures in 96- or 12-well plates were exposed to white light (400-700nm, 1000 lux) or 5mM sodium azide. Their viability (MTT procedure) and production of reactive oxygen species (ROS) were analysed after 24-48 hours. Cultures were also processed for DNA breakdown (TUNEL method) and for certain antigens by immunocytochemistry and electrophoresis/western blotting after exposure to light for 48-96 hours.

Results: : Results: Light and sodium azide caused a loss of cell viability and a stimulation of ROS and breakdown of DNA. The negative effect of light was attenuated by necrostatin-1 and not by the caspase inhibitor zVAD-fmk. In contrast, the influence of sodium azide was unaffected by necrostatin-1 but inhibited by zVAD-fmk. Moreover, light caused an activation of apoptosis inducing factor (AIF), c-Jun, JNK and HO-1 but did not affect alpha fodrin or caspase-3 content. Sodium azide, on the other hand activated of alpha fodrin, caused a stimulation of caspase-3 but did not influence basal expression of AIF, c-Jun, JNK or HO-1. Light, unlike sodium azide also caused p-c-Jun to be translocated from the nucleus to the cytoplasm.

Keywords: ganglion cells • mitochondria • cell survival 
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