The role of S182 and E5-1 on AB secretion in Alzheimer's Disease.

阿尔茨海默病研究的最新进展揭示了由β组成的老年斑淀粉样蛋白作为阿尔茨海默病出现神经原纤维缠结和神经元细胞损失之前的最早事件的重要性。此外，遗传学研究发现了与常见阿尔茨海默病相关的4个致病基因，分别是突变体betaAPP、载脂蛋白E4、S182（早老素-1:PS-1）和E5-1（早老素-2:PS-2）。在本研究中，我们计划1)对2例熟悉的AD病例进行基因分析；2)克隆PS-1野生型，构建突变型PS-1和PS-2；3)制备PS-1和PS-2特异性抗体；4)表达细胞系的建立；5)蛋白水解片段检测及β分泌量测定；6) AD和对照组脑免疫染色和免疫印迹分析；7)表达突变体PS-1的转基因小鼠分析。在PS-1 A260V和PS-1 A285V家族中发现了广泛的老年斑和淀粉样血管病。克隆了野生型PS-1和PS-2、突变型PS-1…More （DELTAM146L、DELTAA246E、DELTAL286V、DELTAExon9、DELTAL392V、DELTAC410Y）和突变型PS-2 （DELTAN141I、DELTAM239V）。鉴定了PS-1 n端（MTELPAPLSYFONAQMSEDNHLS,HSN-2）和c端（LVQPFMDQLAFHQFYI,HS-C）的抗体。瞬时和稳定表达PS-1的细胞系在野生和突变细胞系中均显示出44/40 KD的PS-1全长，26/25 KD的n端片段（NTFs）和c端片段（CTFs）。这些PS-1及其片段无显著性差异。表达betaAPPDELTANL和突变体PS-1的双转染细胞系分泌Abeta1-40和Abeta1-42的量增加。在AD和对照脑中，26/25 KD NTFs和17/16KD CTFs被识别。未观察到全长PS-1。表达DELTAM146L和deltam410y的转基因小鼠在脑内显示出一定分子大小的PS-1及其片段。虽然HSN-2和HS-C在对照脑中标记神经元和过程，但HS-C广泛标记神经原纤维缠结和卷曲纤维，这表明AD脑中PS-1和细胞骨架异常密切相关。少

Recent advance of Alzheimer research revealed the importance of senile plaque amyloids consisting of Abeta as the earliest event of Alzhiemer's disease before appearance of neurofibrillary tangles and neuronal cell loss. Furthermore, genetical study found 4 causal genes linked to familiar Alzheimer's disease, that are, mutant betaAPP,Apolipoprotein E4, S182 (presenilin-1 : PS-1) and E5-1 (presenilin-2 : PS-2). In this study, we planned 1) gene analysis of 2 familiar AD cases ; 2)cloning of wild type of PS-1 and constructing mutant type PS-1 and PS-2 ; 3) making specific antibodies to PS-1 and PS-2 ; 4)establishment of expressing cell lines ; 5)detection of proteolytic fragments and measurement of amount of Abeta secretion ; 6) immunostain and immunoblot analysis of the AD and control brains ; and 7)analysis of transgenic mice expressing mutant PS-1.Extensive senile plaques and amyloid angiopathy were recognized in PS-1 A260V and PS-1 A285V families. Wild type PS-1 and PS-2, mutant PS-1 … More (DELTAM146L,DELTAA246E,DELTAL286V,DELTAExon9, DELTAL392V,DELTAC410Y) and mutant PS-2 (DELTAN141I,DELTAM239V) were cloned. Antilbodies to N-terminus of PS-1 (MTELPAPLSYFONAQMSEDNHLS,HSN-2) and C-terminus of PS-1(LVQPFMDQLAFHQFYI,HS-C) were characterized. Transient and stable cell lines expressing PS-1 showed that 44/40 KD full length of PS-1,26/25 KD N-terminal fragments (NTFs) and C-terminal fragments (CTFs) in both wild and mutant cell lines. No significant difference was pbserved in these PS-1 and their fragments. Doubletransfection cell lines expressing betaAPPDELTANL and mutant PS-1 secreted increased amounts of Abeta1-40 and Abeta1-42. In the AD and control brains, 26/25 KD NTFs and 17/16KD CTFs were recognized. Full length PS-1 was not observed. Transgenic mice expressing DELTAM146L and DELTAC410Y showed the some molecule size PS-1 and their fragmentsin the brains. Although HSN-2 and HS-C labeled neurons and processes in the control brains, neurofibrillary tangles and curly fibers were extensively labeled by HS-C suggesting close relationship with PS-1 and cytoskeletal abnormalities in the AD brains. Less