Membrane Properties, Sunaptic Currents, and Intracellular Ca Transients in Transplanted Neurons in the Model Rats of Diseases of the Central Nervous System

中枢神经系统疾病模型大鼠移植神经元的膜特性、突触电流和细胞内 Ca 瞬变

• 批准号: 07680897
• 负责人: FUKUDA Atsuo
• 金额: $ 1.47万
• 依托单位: Nagoya City University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1995
• 资助国家: 日本
• 起止时间: 1995 至 1996
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07680897/
• 关键词: Parkinson disease; Cerebral ischemia; 3-Nitropropionic acid; Intracellular Ca; Cerebral Cortex; Striatum; Neural transplantation; Cell death; 脳移植; シナプス; 黒質; パッチクランプ

(1) Preparing Model Rats of Diseases of the Central Nervous System : Hemiparkinsonian disease model rats were made with unilateral injection of 6-OHDA to substantia nigra. Cerebral ischemia was performed by unilateral mid cerebral artery occlusion in adult rats and by unilateral common carotid artery ligation with hypoxia (8% oxygen) in immature rats. 3-Nitropropionic acid was administered intraperitoneously to induce Huntington diesease-like symptoms and striatal lesions in rats.(2) In Vitro Model of the Diseases Described Above : Cerebralischemia was simulated by oxygen-glucose deprivation of neocortical slices. LayrII/III pyramidal cells were more vulnerable to Ca^<2+>-overload than those in the other layrs due to NMDA receptor susceptibility. We also found that GABA may aggravate ischemic damage directly and/or via mediation of glutamate-NMDA excitotoxicity as a result of E_<el> shift during ischemia. Perfusion of cultured striatal and cortical glial cells with 3-nitropropionic aci … More d caused cellular Ca^<2+>-overload leading to necrotic deathby reverse operation of Na^+-Ca^<2+> exchanger.Physiological Experiments on Transplanted Neurons in Brain Slices from the Model Rats : We established methods for labeling and identification of living donor cells in brains slices of recipient himiparkinsonian and cerebral ischemia model rats. Voltage dependent Na^+ and K^+ currents as well as glutamate-induced intracellular Ca^<2+> increase were successfully recorded from the grafted cells identified by the labeling. Interconnection between the host and grafted cells was functionally proved in brain slices as intracellular Ca^<2+> transients in response to the electrical stimulation of neighboring host tissue.Detection of Gene Expression Induced in the Model Rats : In the 3-nitropropionic acid-induced Huntington disease-like striatal lesion, expression of iNOS mRNA increased as revealed by RT-PCR method Immunohistochemical staining also showed an increased level of iNOS in parallel with mRNA. Less

(1)中枢神经系统疾病模型大鼠的制备：通过单侧黑质注射6-OHDA制成偏侧帕金森病模型大鼠。采用成年大鼠单侧大脑中动脉闭塞法和幼年大鼠单侧颈总动脉结扎缺氧（8%氧）法制备脑缺血模型。3-腹腔注射硝基丙酸诱导大鼠出现亨廷顿病样症状和纹状体病变。(2)上述疾病的体外模型：通过新皮层切片的氧-葡萄糖剥夺来模拟脑缺血。由于NMDA受体的敏感性，第II/III层锥体细胞比其他层锥体细胞更易受到Ca^<2+>超载的影响。我们还发现GABA可直接或通过介导缺血时E_移位引起的谷氨酸-NMDA兴奋毒性而加重缺血性损伤<el>。用3-硝基丙酸灌注培养的纹状体和皮质胶质细胞 ...更多信息 d通过Na^+-Ca^&lt;2 +&gt;交换器的反向操作引起细胞Ca^&lt;2 +&gt;超载，导致坏死性死亡。模型大鼠脑片移植神经元的生理学实验：建立了帕金森病和脑缺血模型大鼠脑片活体供者细胞的标记和鉴定方法。从标记鉴定的移植细胞中成功记录到电压依赖性Na^+和K^+电流以及谷氨酸诱导的细胞内Ca^2+增加。在脑切片中，宿主细胞和移植细胞之间的相互连接在功能上被证明为响应于邻近宿主组织的电刺激的细胞内Ca^&lt;2+&gt;瞬变。在3-硝基丙酸诱导的亨廷顿病样纹状体病变中，RT-PCR法显示iNOS mRNA表达增加，免疫组化染色显示iNOS表达水平与mRNA水平平行增加。少

项目成果

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Nishino, H. et al.: "3-Nitropropionic acid produces striatum selective lesions accompanied by iNOS expression." Journal of Chemical Neuroanatomy. 10. 209-212 (1996)

Nishino, H. 等人：“3-硝基丙酸产生纹状体选择性损伤，并伴有 iNOS 表达。”