Nitric oxide mediates excitotoxic and anoxic damage in rat retinal ganglion cells cocultured with astroglia.
BACKGROUND
Nitric oxide has been implicated in the process of retinal ganglion cell death in glaucoma.


OBJECTIVE
To investigate the role of nitric oxide in mediating retinal ganglion cell death in a culture system that models glial-neuronal interactions at the level of the optic nerve head.


METHODS
Dissociated retinal ganglion cells from neonatal rats were plated on monolayers of astroglia and identified by retrograde labeling with the fluorescent marker 1.1-dioctadecyl-,3,3,3,tetramethylindocarbocyanineperchlorate. Two days after dissociation, cocultures of retinal ganglion cells and glia were treated with graded concentrations of the nitric oxide synthase inhibitor N-nitro-L-arginine (NNA), and exposed to either anoxia for 1 to 24 hours or excitatory amino acids for 6 hours. Surviving retinal ganglion cells were counted with fluorescence microscopy and expressed as a percentage of retinal ganglion cells surviving in control cultures.


RESULTS
Cell survival after anoxia increased in a dose-dependent fashion with exposure to NNA. Mean +/- SD survival rate of retinal ganglion cells after 6 hours of anoxia was 57%+/-10% with NNA treatment compared with 31%+/-3% without treatment (P<.01). When treated with excitatory amino acids, cell survival was 31%+/-6% after administration of N-methyl D-aspartate, 500 micromol/L, and 27%+/-8% after administration of sodium glutamate, 500 micromol/L. Survival was increased in cultures with exposure to NNA, 100 micromol/L, to 53%+/-11% and 69%+/-11%, respectively (P<.01).


CONCLUSIONS
In this coculture of retinal ganglion cells and astroglia, reduction of the glial source of nitric oxide through nitric oxide synthase inhibition provided partial but significant protection against the lethal effects of anoxia and excitatory amino acids on retinal ganglion cells.


CONCLUSIONS
Neuroprotective agents may play a role in patients with glaucoma who have progressive visual field loss, despite satisfactory control of intraocular pressure. Inhibition of nitric oxide synthase at the level of the optic nerve head may contribute to a clinically significant level of neuroprotection.
Morgan
J
J
Cardiff Eye Unit, University Hospital of Wales.
Caprioli
J
J
Koseki
Y
Y
D000818:Animals; D001253:Astrocytes / Q000187:drug effects / Q000201:enzymology / Q000473:pathology*; D016923:Cell Death; D015687:Cell Hypoxia; D002470:Cell Survival; D002478:Cells, Cultured; D018920:Coculture Techniques; D004305:Dose-Response Relationship, Drug; D004791:Enzyme Inhibitors / Q000494:pharmacology; D018690:Excitatory Amino Acid Agonists / Q000494:pharmacology*; D018698:Glutamic Acid / Q000494:pharmacology; D016202:N-Methylaspartate / Q000494:pharmacology; D009569:Nitric Oxide / Q000502:physiology*; D019001:Nitric Oxide Synthase / Q000037:antagonists & inhibitors; D019335:Nitroarginine / Q000494:pharmacology; D051381:Rats; D017207:Rats, Sprague-Dawley; D012165:Retinal Ganglion Cells / Q000187:drug effects / Q000473:pathology*
D004791:Enzyme Inhibitors; D018690:Excitatory Amino Acid Agonists; D019335:Nitroarginine; D009569:Nitric Oxide; D018698:Glutamic Acid; D016202:N-Methylaspartate; D019001:Nitric Oxide Synthase