Neuropathological study on Abeta toxicity using transgenic mice with human APP

使用人APP转基因小鼠进行Abeta毒性的神经病理学研究

• 批准号: 11680742
• 负责人: MORI Hiroshi
• 金额: $ 2.37万
• 依托单位: Osaka 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: 1999
• 资助国家: 日本
• 起止时间: 1999 至 2000
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-11680742/
• 关键词: Alzheimer's disease; APP; Abeta, senile plaque; transgenic mouse; model; neurotoxicity; presenilin; プレセニリン; マルツハイマー病

Amyloid plaques and neurofibrillary tangles are neuropathological and molecular hallmarks for Alzheimer's disease (AD). Their presence is highly important to understand the etiology of the disease. Making model mice is undoubtedly crucial to examine the molecular mechanism how to develop plaque formation and cell death in cerebral tissues and to study a potent therapeutic drug. We tried to make a mouse that bears cerebral amyloid plaques by cross the mutated APP717 (referred to as London mutation) mouse and the mutated PS1 mouse. Both mutations were generated by PCR with human APP and PS1 cDNA templates. These two DNA constructs for transgenic mice were integrated in prion promoter plasmid vector (from Dr. D.Borchelt, Jonhs Hopkins Univ.). We succeeded in generate three lines of mice with human APP717 and two lines of mice with human mutated PS1. The lines were established to show their genetic and protein expression. Amyloid protein was exmained and established to show neurotoxicity using fetal hippocampal neurons derived from these mice. An antagonic effect was found to be associated with crystamine G that is an analogue of congo red to bind amyloid fibril in AD brain.

淀粉样蛋白斑和神经原纤维缠结是阿尔茨海默病 (AD) 的神经病理学和分子标志。它们的存在对于了解该疾病的病因非常重要。制作小鼠模型无疑对于研究脑组织斑块形成和细胞死亡的分子机制以及研究有效的治疗药物至关重要。我们试图通过突变的APP717（称为伦敦突变）小鼠与突变的PS1小鼠杂交，制备出具有脑淀粉样斑块的小鼠。这两种突变都是使用人类 APP 和 PS1 cDNA 模板通过 PCR 产生的。这两种转基因小鼠的 DNA 构建体被整合到朊病毒启动子质粒载体（来自约翰·霍普金斯大学 D.Borchelt 博士）中。我们成功地产生了三个带有人类 APP717 的小鼠品系和两个带有人类突变 PS1 的小鼠品系。建立这些细胞系是为了显示它们的遗传和蛋白质表达。使用源自这些小鼠的胎儿海马神经元检查并确定淀粉样蛋白以显示神经毒性。研究发现晶胺 G 具有拮抗作用，晶胺 G 是刚果红的类似物，可与 AD 脑中的淀粉样原纤维结合。

项目成果

Tamaoka, A., Miyatake, F., Matsuno, S., Ishii, K., Nagase, S., Sahara, N., Ono, S., Mori, H., Wakabayashi, K., Tsuji, S., Takahashi, H.& Shoji, S.: "Apolipoprotein E allele-dependent antioxidant activity in brains with Alzheimer's disease"Neurology. 54. 2

玉冈，A.，宫武，F.，松野，S.，石井，K.，长濑，S.，撒原，N.，小野，S.，森，H.，若林，K.，辻，S.，

Sahara,N.,Tomiyama,T.,& Mori,H.: "Missense point mutations of tau to segregate with FTDP-17 exhibit site-specific effects on microtubule structure in COS cells : A novel action of R406W mutation"J.Neurosci.Res.. 60(3). 380-387 (2000)

撒哈拉，北，富山，T.，

Arai,T., Akiyama,H., Ikeda,K., Kondo,H., & Mori,H.: "Immunohistochemical localization of amyloid beta-protein with amino-terminal aspartate in the cerebral cortex of patients with Alzheimer's disease"Brain Res.. 823(1-2). 202-206 (1999)

荒井T.、秋山H.、池田K.、近藤H.、

Mori, H.: "Untagling Alzheimer's disease from fibrous lesions of neurofibrillary tangles and senile plaques"Neuropathology. 20. S55-S60 (2000)

Mori, H.：“从神经原纤维缠结和老年斑的纤维病变中解开阿尔茨海默病”神经病理学。

Murayama,K., Osami,M.,Kametani,F.,Tomita,T.,Mori,H., et al.: "Molecular interactions between presenilin and calpain : Inhibition of m-calpain protease activity by presenilin-1,2 and cleavage of presenilin-1 by m-, u-calpain"Int.J.Molecular Medicine. 5. 26

Murayama,K.、Osami,M.、Kametani,F.、Tomita,T.、Mori,H. 等人：“早老素和钙蛋白酶之间的分子相互作用：早老素-1,2 对 m-钙蛋白酶活性的抑制