A study on the cell death of primary cultured rat's cortical neurons induced by exicitatory amino acids and protective strategies against it.

兴奋性氨基酸诱导原代培养大鼠皮层神经元细胞死亡及其保护策略的研究

• 批准号: 04670701
• 负责人: OKAMOTO Motoi
• 金额: $ 1.28万
• 依托单位: OKAYAMA UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1992
• 资助国家: 日本
• 起止时间: 1992 至 1993
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-04670701/
• 关键词: cell death; excitatory amino acids; cell culture; cortical neuron; proteoglycan; chondroitin sulfate; heparan sulfate; ELISA; 培養神経細胞; ラット; 大脳皮質; グルタミン酸; 培養細胞; 神経細胞死; 無血清培養; ミクロデキマトリン; コンドロイチン硫酸プロテオグリカン; グリタミン酸

In this project, we established an in vitro model system for studying the mechanism of neuronal death by using primary cultured rat's cortical neurons, and examined the effects of extracellular matrix molecules on the cell death induced by excitatory amino acids.1)The death of primary cultured cortical neurons of rat fetuses increased in parallel with developmental stage in vivo, providing the fact that primary cultured neurons can be a in vitro model for investing developmental changes of neuronal sensitivity to excitatory amino acids.2)Chondroitin sulfate proteoglycans (CSPG)prepared from neonatal rat's brain protected neuronal death induced by 24 h exposure to glutamate.3)The protective effect of CSPG was due to its core protein, but not due to glycosaminoglycan side chains nor adsorption of glutamate.4)The protective action of CSPG was comparable to a NMDA antagonist, MK-801, and was stronger than AMPA antagonist, NBQX.5)CSPG also protected delayd neuronal death following 10 min. exposure to glutamate, as well as kainate, NMDA,and AMPA.6)A enzyme-linked immunosolvent assay for measuring nanogram level of heparan sulfate proteoglycan was established by using lipoprotein lipase (LPL) and anti-LPL rabbit's serum.Based on these findings, we suggested a possible involvement of CSPG in the neuronal death in acute pathological conditions such as ischemia, hypoxia and in chronic degenerative diseases.

本项目利用原代培养的大鼠皮层神经元，建立了研究神经元死亡机制的体外模型系统，并考察了细胞外基质分子对兴奋性氨基酸诱导的细胞死亡的影响。1）原代细胞死亡培养的大鼠胎儿皮质神经元与体内发育阶段平行增加，这提供了原代培养神经元可以作为研究神经元对兴奋性氨基酸敏感性发育变化的体外模型的事实。 2）软骨素从新生大鼠脑中制备的硫酸蛋白聚糖（CSPG）可以保护24小时接触谷氨酸引起的神经元死亡。3）CSPG的保护作用是由于其核心蛋白，而不是由于糖胺聚糖侧链或谷氨酸的吸附。4） CSPG 的保护作用与 NMDA 拮抗剂 MK-801 相当，并且强于 AMPA 拮抗剂 NBQX。5)CSPG 还具有保护作用10分钟后神经元延迟死亡。暴露于谷氨酸盐、红藻氨酸、NMDA和AMPA。6)使用脂蛋白​​脂肪酶(LPL)和抗LPL兔血清，建立了一种酶联免疫溶剂测定法，用于测量纳克级硫酸乙酰肝素蛋白聚糖。 ，我们认为 CSPG 可能参与急性病理状况（例如缺血、缺氧）和慢性退行性疾病中的神经元死亡。

项目成果

Okamoto Motoi 他: "Excitotoxic death of cultured cortical neurons:developmental changes and prophylactic effect of chondroitin sulfate proteoglycans." Neurosciences. 19. 163-172 (1993)

Okamoto Motoi 等人：“培养的皮质神经元的兴奋毒性死亡：硫酸软骨素蛋白聚糖的发育变化和预防作用。神经科学”19. 163-172 (1993)。

Okamoto Motoi 他: "Utilization of a serum-free primary culture of cortical neurons by using cyclodextrins in neurobiological research." 岡山大学医療技術短期大学部紀要. (in press). (1994)

Okamoto Motoi 等人：“在神经生物学研究中使用环糊精来利用皮质神经元的无血清原代培养物。”冈山大学医学技术学院通报（1994 年）。

Okamoto Motoi 他: "A protective action of chondroitin sulfate proteoglycans against neuronal cell death induced by glutainate" Brain Research. (in press). (1994)

Okamoto Motoi 等人：“硫酸软骨素蛋白聚糖对谷氨酸诱导的神经元细胞死亡的保护作用”《大脑研究》（出版中）。