Akaike A, Adachi K, Kaneda K
Department of Pharmacology, Graduate School of Pharmaceutical Sciences, Kyoto University, Japan.
Nihon Yakurigaku Zasshi. 1998 Feb;111(2):97-104. doi: 10.1254/fpj.111.97.
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.
本综述描述了评估兴奋性氨基酸和短暂性缺血诱导的视网膜神经变性的技术。在培养的大鼠皮质细胞中检测了谷氨酸诱导的神经毒性。从胎鼠视网膜获得的培养物在补充有10%胎牛血清或10%马血清的伊格尔最低必需培养基中，于37℃、5%二氧化碳饱和湿度的气氛中培养10 - 14天。通过台盼蓝排斥法对谷氨酸诱导的神经毒性进行定量。培养物暴露于谷氨酸10分钟，然后在无谷氨酸培养基中孵育1小时后，其活力显著降低。在成年大鼠中检测了N - 甲基 - D - 天冬氨酸（NMDA）诱导的视网膜损伤。通过视盘的视网膜横切片用苏木精和伊红染色。单次玻璃体内注射NMDA在注射7天后损伤神经节细胞层和内网状层，而不影响其他视网膜层。通过置于前房的针将眼压升高45分钟诱导视网膜缺血。缺血诱导的视网膜损伤被MK - 801抑制。这些结果表明，本综述中描述的技术可用于开发对视网膜缺血期间发生的神经变性具有神经保护作用的新药。
