April 2009
Volume 50, Issue 13
Free
ARVO Annual Meeting Abstract  |   April 2009
Brimonidine Is Neuroprotective Against Glutamate-Induced Neurotoxicity, Oxidative Stress and Hypoxia in Purified Rat Retinal Ganglion Cells
Author Affiliations & Notes
  • K. Y. Lee
    Glaucoma Service, Singapore National Eye Centre, Singapore, Singapore
    Ophthalmology, University of Tokyo, Graduate School of Medicine, Tokyo, Japan
  • M. Nakayama
    Ophthalmology, University of Tokyo, Graduate School of Medicine, Tokyo, Japan
  • M. Aihara
    Ophthalmology, University of Tokyo, Graduate School of Medicine, Tokyo, Japan
  • Y. Chen
    Ophthalmology, University of Tokyo, Graduate School of Medicine, Tokyo, Japan
  • M. Araie
    Ophthalmology, University of Tokyo, Graduate School of Medicine, Tokyo, Japan
  • Footnotes
    Commercial Relationships  K.Y. Lee, None; M. Nakayama, None; M. Aihara, None; Y. Chen, None; M. Araie, None.
  • Footnotes
    Support  None.
Investigative Ophthalmology & Visual Science April 2009, Vol.50, 109. doi:
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      K. Y. Lee, M. Nakayama, M. Aihara, Y. Chen, M. Araie; Brimonidine Is Neuroprotective Against Glutamate-Induced Neurotoxicity, Oxidative Stress and Hypoxia in Purified Rat Retinal Ganglion Cells. Invest. Ophthalmol. Vis. Sci. 2009;50(13):109.

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

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Abstract

Purpose: : To determine the neuroprotective effect of 2-adrenergic agonist, brimonidine, in the presence of glutamate-induced neurotoxicity, oxidative stress and hypoxia on in-vitro cultures of purified rat retinal ganglion cells (RGCs).

Methods: : Purified RGCs cultures were obtained from retinas of 6 to 8 day old Wistar rats, following a two-step immuno-panning procedure. After 72 hours of cultivation, the neuroprotective effect of brimonidine (0.01µM, 0.1µM and 1 µM) was investigated by culturing the RGCs in glutamate, oxidative and hypoxic stress for a further 72 hours, 24 hours and 12 hours respectively. Glutamate neurotoxicity was induced by adding glutamate (25µM), while for oxidative stress, culture medium with B27 supplement without anti-oxidants were substituted. Hypoxic stress was induced by cultivation in controlled-atmosphere incubator with oxygen levels of 5% normal partial pressure. The RGC viability in each condition normalized to that under normal condition was evaluated as live cell percentage based on a total of 7-8 experiments.

Results: : The cell survival percentage of control cultures exposed to glutamate, oxidative and hypoxic stress were 58.2%, 59.3% and 53.2% respectively. Brimonidine (1 µM) significantly increased RGC survival in the presence of glutamate (80.6%), oxidative (79.8%) and hypoxic (77.4%) stress (P=0.0018, <0.0001 and <0.0001 respectively). In the presence of 2-adrenergic antagonist yohimbine (10 µM), RGC viability with brimonidine (1 µM) was not significantly different from controls.

Conclusions: : Brimonidine increased survival of purified rat RGCs in the presence of glutamate neurotoxicity, oxidative and hypoxic stress. The neuroprotective effect of brimonidine is mediated via 2-adrenergic receptors at the RGC level.

Keywords: neuroprotection • retinal culture • apoptosis/cell death 
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