Development of animal models of human neurodegenerative diseases using transgenic mice

使用转基因小鼠开发人类神经退行性疾病动物模型

• 批准号: 07558106
• 负责人: NAGATSU Toshiharu
• 金额: $ 10.94万
• 依托单位: Fujita Health University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (A)
• 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-07558106/
• 关键词: transgenic mice; immunotoxin; conditional disruption; neurodegeneration; noradorenergic neurons; autonomic nerve failure; Parkinson's disease; Huntington's disease

We development a transgenic approach, termed immunotoxin-mediated cell targeting (IMCT). Transgenic mice were created that express the human interleukin 2 receptor alpha subunit (IL-2Ralpha) in a specific cell type. and the cells were ablated based on the cytotoxicity of a recombinant immunotoxin anti-Tac (Fv) -PE40. DIL transgenic mice were created that express IL-2Ralpha in noradrenergic neurons under the control of the dopamine beta-hydroxylase (DBH) gene promoter. The DIL mice were treated intracerebroventricularly with the immunotoxin, and showed a characteristic behavioral abnormality, accompanied by a loss of DBH-cantaining neurons and a significant decrease in DBH activity and noradrenaline levels in various regions of the brain. When the DIL mice were treated intravenously with the immunotoxin, the sympathetic nerves were degenerated, accompanied by decreases in tissue catecholamine levels, motor activity, body weigh, body temperature, and heart rate, and by abnormal electrocr … More adiogram. The DIL transgenic mice could be the animal models of autonormic nerve failure. The IMCT method was also applied for producing animal models of Parkinson's disease and Huntington's disease. For the animal model of Parkinson's disease, transgenic mice were created that express IL-2Ralpha in dopaminergic neurons in the striatum under the control of the tyrosine hydroxylase (TH) gene promoter. For the animal model of Huntingtons's disease, knockout mice were created that express IL-2Ralpha gene in the locus of dopamine D2 receptor gene by gene targeting. Immunotoxin was injected stereotaxically into the striatum to ablapt either dopamine neurons or GABA neurons. IMCT method was also applied to the negative selection in mouse embryonic stem (ES) cells. By immunotoxin treatment, only ES cells bearing human IL-2Ralpha were efficiently eliminated, indicating that IMCT can be used as a novel strategy for positive and negative selection to enrich ES cell clones with a targeted mutation. Less

我们开发了一种转基因方法，称为免疫毒素介导的细胞靶向 (IMCT)，其在特定细胞类型中表达人白细胞介素 2 受体 α 亚基 (IL-2Ralpha)，并根据细胞毒性消除细胞。构建了重组免疫毒素抗 Tac (Fv)-PE40 转基因小鼠，其在去甲肾上腺素能神经元中表达 IL-2Rα。将多巴胺β-羟化酶（DBH）基因启动子注射到脑室内的DIL小鼠表现出特征性的行为异常，伴随着大脑区域中含有DBH的神经元的损失以及DBH活性和去甲肾上腺素水平的显着降低。当 DIL 小鼠接受免疫毒素静脉注射时，交感神经发生退化，并伴有组织儿茶酚胺水平、运动能力下降。 IMCT方法也可用于制作帕金森病和亨廷顿病的动物模型。对于帕金森病的动物模型，创建了转基因小鼠，在酪氨酸羟化酶 (TH) 基因的控制下，纹状体中的多巴胺能神经元表达 IL-2Rα对于亨廷顿病的动物模型，通过基因靶向将免疫毒素注射到纹状体中以消除多巴胺神经元或GABA神经元，从而创建在多巴胺D2受体基因座中表达IL-2Rα基因的敲除小鼠。该方法也适用于小鼠胚胎干（ES）细胞的阴性选择。通过免疫毒素处理，只有携带人IL-2Rα的ES细胞才有效。消除，表明 IMCT 可用作正向和负向选择的新策略，以通过靶向突变富集 ES 细胞克隆。

项目成果

Kazuto Kobayashi, Ira Pastan and Toshiharu Nagatsu: Controlled genetic ablation by immunotoxin-mediated cell targeting Transgenic Animals-Generation and Use, ed.by L.M.Houdebine, Harwood Academic Publishers. Amsterdam, (1997)

Kazuto Kobayashi、Ira Pastan 和 Toshiharu Nagatsu：通过免疫毒素介导的细胞靶向转基因动物的控制基因消融 - 生成和使用，L.M.Houdebine 编辑，哈伍德学术出版社。

K.Kobayashi: "A novel stratagy for the negative selection in mouse embryonic stem cells operated with immunotoxin-mediated cell targeting" Nucleic Acids Research. 24. 3653-3655 (1996)

K.Kobayashi：“通过免疫毒素介导的细胞靶向操作的小鼠胚胎干细胞负选择的新策略”核酸研究。

K.Kobayashi: "Transgenic Animals : Generation and Use" Harwood Academic Publishers, (1997)

K.Kobayashi：“转基因动物：生成和使用”哈伍德学术出版社，（1997）

Kazuto Kobayashi, Tamae Ohye, Ira Pastan, and Toshiharu Nagatsu: "A novel strategy for the negative selection in mouse embryonic stem cells operated with immunotoxin-mediated cell targeting" Nucleic Acids Research. Vol.24, No.18. 3653-3655 (1996)

Kazuto Kobayashi、Tamae Ohye、Ira Pastan 和 Toshiharu Nagatsu：“通过免疫毒素介导的细胞靶向操作的小鼠胚胎干细胞负选择的新策略”核酸研究。

K.Kobayashi: "A novel stratagy for the negative selection in mouse embryonic stem cells operated with immunotoxin-mediated cell targeting" Nucleic Acids Research. 24・18. 3653-3655 (1996)

K.Kobayashi：“通过免疫毒素介导的细胞靶向操作进行小鼠胚胎干细胞负选择的新策略”，《核酸研究》24·18（1996）。