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)的神经病理和分子特征。它们的存在对于了解疾病的病因是非常重要的。制作小鼠模型无疑是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的关键。我们试图通过将突变的APP717(简称伦敦突变)小鼠与突变的PS1小鼠杂交，制造出一只带有大脑淀粉样斑块的小鼠。这两个突变都是通过用人APP和PS1cDNA模板进行聚合酶链式反应而产生的。将这两个转基因小鼠的DNA构建载体整合到PrP启动子载体中(来自约翰霍普金斯大学D.Borchelt博士)。我们成功地培育了三株携带人APP717基因的小鼠和两株携带人突变PS1基因的小鼠。这些品系的建立是为了显示它们的遗传和蛋白质表达。利用来自这些小鼠的胎鼠海马神经元，对淀粉样蛋白进行了研究，并证实了其具有神经毒性。研究发现，结晶胺G具有抗炎作用，它是一种刚果红的类似物，可以结合AD大脑中的淀粉样原纤维。

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.