[Techniques for evaluating neuronal death of the retina in vitro and in vivo].
This review describes the techniques to evaluate retinal neurodegeneration induced by excitatory amino acids and transient ischemia. Glutamate-induced neurotoxicity was examined in cultured rat cortical cells. Cultures obtained from the retinas of fetal rats were incubated in Eagle's minimal essential medium supplemented with 10% fetal calf serum or 10% horse serum at 37 degrees C in a humidified 5% CO2 atmosphere for 10-14 days. The neurotoxicity induced by glutamate was quantified by trypan blue exclusion. The viability of cultures was markedly reduced by a 10-min exposure to glutamate followed by incubation with glutamate-free medium for 1 hr. N-methyl-D-aspartate (NMDA)-induced retinal damage was examined in adult rats. Transverse sections of the retinas through the optic disk were stained with hematoxylin and eosin. A single intravitreal injection of NMDA damaged the ganglion cell layer and the inner plexiform layer without affecting the other retinal layers 7 days after injection. Retinal ischemia was induced by elevating the intraocular pressure for 45 min through the needle which was placed in the anterior chamber. Ischemia-induced retinal damage was inhibited by MK-801. These results indicate that the techniques described in this review can be employed to develop new drugs possessing neuroprotective action against neurodegeneration that occurs during retinal ischemia.
Akaike
A
A
Department of Pharmacology, Graduate School of Pharmaceutical Sciences, Kyoto University, Japan.
Adachi
K
K
Kaneda
K
K
D000818:Animals; D016923:Cell Death; D002478:Cells, Cultured; D016291:Dizocilpine Maleate / Q000627:therapeutic use; D018691:Excitatory Amino Acid Antagonists / Q000627:therapeutic use; D005901:Glaucoma / Q000188:drug therapy / Q000209:etiology; D005971:Glutamates / Q000633:toxicity; D006801:Humans; D007267:Injections; D007511:Ischemia / Q000188:drug therapy / Q000209:etiology; D016202:N-Methylaspartate / Q000633:toxicity; D009474:Neurons / Q000187:drug effects; D018696:Neuroprotective Agents / Q000627:therapeutic use; D051381:Rats; D012160:Retina / Q000166:cytology* / Q000187:drug effects; D012171:Retinal Vessels; D014822:Vitreous Body
D018691:Excitatory Amino Acid Antagonists; D005971:Glutamates; D018696:Neuroprotective Agents; D016202:N-Methylaspartate; D016291:Dizocilpine Maleate