Neuropathological study on Abeta toxicity using transgenic mice with human APP

淀粉样斑块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特征。它们的存在对了解该病的病因非常重要。制作模型小鼠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无疑是至关重要的。我们试图通过将突变的APP717（称为伦敦突变）小鼠与突变的PS1小鼠杂交，制造出携带脑淀粉样蛋白斑块的小鼠。用人APP和PS1 cDNA模板进行PCR，得到两个突变体。这两种用于转基因小鼠的DNA构建被整合到朊病毒启动子质粒载体中（来自Dr. D.Borchelt, johns Hopkins university）。我们成功地培育了3株携带人类APP717的小鼠和2株携带人类突变PS1的小鼠。建立了这些细胞系，以显示它们的遗传和蛋白质表达。利用取自这些小鼠的胎儿海马神经元对淀粉样蛋白进行了检测，并证实其具有神经毒性。发现一种对抗作用与晶体胺G有关，晶体胺G是一种类似于刚果红的物质，可以结合AD大脑中的淀粉样蛋白纤维。

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.