FRET对活体细胞内APP裂解过程的动态研究
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摘要
目的
构建含有突变型APP的荧光真核表达系统以研究APP的酶解过程和Aβ产生的分子机制。
方法
以质粒pcDNA3.0-CFP-CaM-YFP的BglII酶切产物为模板,通过聚合酶链式反应(PCR)分别得到编码蓝色荧光蛋白(cyan fluorescence protein, CFP)和黄色荧光蛋白碱基序列(yellow fluorescence protein, YFP);以pcDNA3.0-APP为模板,通过聚合酶链式反应(PCR)得到含有APP717突变的最后300碱基片段(C99);生物合成含有Swedish突变的APP中间54个碱基片段(54bp)。利用基因工程技术将CFP、54bp、YFP、C99片段克隆至载体质粒pcDNA3.0中,得到重组质粒pcDNA3.0-CFP-54bp-YFP-C99和pcDNA3.0-CFP-54bp-YFP,酶切和测序鉴定。将构建的融合基因转染至SH-SY5Y细胞中,通过RT-PCR检测融合基因mRNA表达,利用多光子共聚焦显微镜观察不同时间点(12h、24h、48h、72h、96h)转染细胞内的荧光蛋白表达情况。细胞转染12h、18h、24h、36h、48h、72h后,检测FRET,予以波长为820nm的CFP激发光激发,采集发射波长为473nm的CFP图像,同时采集发射波长为525nm的YFP图像;计算CFP、YFP荧光强度(ICFP和IYFP)和两者的比率(Fret=IYFP/ICFP),绘制曲线。pcDNA3.0-CFP-54bp-YFP-C99转染细胞48h后,利用多光子共聚焦显微镜观察细胞内黄色荧光蛋白的表达、分布以及YFP标记的Aβ去向。免疫细胞化学鉴定Aβ的生成。pcDNA3.0-CFP-54bp-YFP-C99、pcDNA3.0-CFP-54bp-YFP转染细胞12h、24h、36h、48h、72h、96h后,MTT检测转染细胞活性,了解Aβ对细胞的影响。
结果
1、CFP、YFP、C99的PCR产物大小分别为818bp、738bp、325bp,经过不同的双酶切得到的酶切产物大小分别为702bp、723bp和310bp,电泳证实与预期一致;重组质粒pcDNA3.0-CFP-54bp-YFP-C99、pcDNA3.0-CFP-54bp-YFP分别经过不同的组合酶切,得到的目的片段长度正确;测序鉴定证明融合基因CFP-54bp-YFP-C99、CFP-54bp-YFP的序列与“gene bank”的碱基信息相符。
2、将转染细胞进行RT-PCR显示有目的片段,分别对应于CFP-54bp-YFP-C99和YFP-C99,对照细胞内则无相应片段。
3、转染细胞内有蓝色(和)黄色荧光表达,在转染12h后已经可以观察到细胞内有荧光分布,24h荧光逐渐增多,至48h荧光进一步增多增强,72h荧光有所减少,96h荧光明显减少减弱。
4、pcDNA3.0-CFP-54bp-YFP细胞在转染后始终存在FRET,细胞轮廓光滑;pcDNA3.0-CFP-54bp-YFP-C99细胞在转染12h后有微弱FRET,以后无FRET,细胞内有颗粒广泛沉积。
5、CFP-54bp-YFP转染细胞Fret不变; CFP-54bp-YFP-C99转染细胞Fret下降。
6、CFP-54bp-YFP-C99在转染细胞48h后,能够被β、γ分泌酶裂解产生Aβ。
7、Aβ产生于首先细胞内,聚集成颗粒,广泛分布于细胞内和细胞膜上,细胞外间隙也有少量分布,但没有形成明显沉积。
8、Aβ形成后细胞形态异常。
9、免疫细胞化学进一步证实Aβ的生成。
10、MTT证实细胞内聚集的Aβ使得细胞活性下降,与对照组相比差异有统计学意义。
结论
1、荧光标记并含有Swedish (和APP717 )突变的重组质粒pcDNA3.0-CFP-54bp-YFP-C99和pcDNA3.0-CFP-54bp-YFP构建成功。
2、融合基因在mRNA和蛋白水平均能够正确表达,为下一步研究APP裂解和Aβ产生提供了一种有力工具。
3、FRET能够敏感准确的检测APP有无发生β裂解。
4、CFP-54bp-YFP-C99能够被β分泌酶裂解而CFP-54bp-YFP不能被裂解,提示C99可能起到信号肽样的引导定位作用,对β分泌酶裂解APP具有重要意义;干扰C99可能会抑制Aβ的产生,本实验为探索AD的早期干预提供一种新思路。
5、融合基因能够正确表达并被裂解,生成YFP标记的Aβ。
6、Aβ产生于细胞内多个部位,并有少量被分泌至细胞外。
7、Aβ在细胞外形成沉积之前,首先在细胞内聚集并产生继发性细胞毒作用,造成细胞活性下降,形态异常。
Objective
To investigate the mechanism of the APP cleavage progress and Aβgeneration, the construction of recombinant eukaryotic expression plasmid was made encoding Swedish and APP717 mutations of amyloid precursor protein (APP) and fluorescent protein.
Methods
The cyan and yellow fluorescence protein sequences (which were named as CFP and YFP, respectively.) were obtained by polymerase chain reaction (PCR) with the template of BglII-digestion product of the plasmid pcDNA3.0-CFP-CaM-YFP. The last 300 bases of APP sequence (which was named as C99 containing APP717 mutation) were amplified by PCR with the template of the plasmid pcDNA3.0-APP harboring APP717 mutation. The 54 bases in the middle of APP sequence were synthesized (which was named as 54bp containing Swedish mutation) by biological company. The four fragments, mentioned above (which were CFP, YFP, C99 as well as 54bp.) were inserted into the vector pcDNA3.0. By genetic engineering the recombinant plasmid pcDNA3.0-CFP-54bp-YFP-C99 and pcDNA3.0-CFP-54bp-YFP were constructed and identified by enzyme digestion and sequencing assay. The recombinant plasmids pcDNA3.0-CFP-54bp-YFP-C99 and pcDNA3.0-CFP-54bp-YFP were transfected into SH-SY5Y cells respectively. The mRNA expression of the fusion gene was examined by reverse transcript polymerase chain reaction assay (RT-PCR). The protein expression of fusion gene and fluorescence change was detected by multi-photon con-focal microscopy at 12h, 24h, 48h, 72h and 96h after transfection. At 12h, 18h, 24h, 36h, 48h and 72h after transfection of pcDNA3.0-CFP-54bp-YFP-C99 or pcDNA3.0-CFP-54bp-YFP, as soon as the cells were excited at 820nm of CFP excitation wavelength, the CFP images were collected at the CFP emission wavelength of 473nm and YFP images were also collected at the YFP emission wavelength of 525nm. FRET phenomenon was observed to get an understanding ofβ-cleavage of APP. Calculate the intensities of CFP (ICFP) and YFP (IYFP) and then turn them into the ratio Fret ( Fret=IYFP : ICFP). At 48h after pcDNA3.0-CFP-54bp-YFP-C99 transfection, multi-photon con-focal microscope was used to detect the yellow fluorescence, to observe its distribution and trace Aβlabeled by YFP. Aβgeneration was confirmed by immunocytochemistry. The viability of transfection cells was measured via MTT assay to find the Aβeffect on the cells at specific time point.
Results
1. The length of PCR-product of CFP、YFP and C99 was 818bp、738bp and 325bp. The length of product of CFP、YFP and C99 via different double-digestion was 702bp、723bp and 310bp. The electrophoresis result is identical to our expectation. The different double-digestion of the plasmid pcDNA3.0-CFP-54bp-YFP-C99 and pcDNA3.0-CFP-54bp-YFP resulted in different fragments and confirmed the accomplishment of the two recombinant plasmids. Finally sequencing assay showed that the fusion genes of CFP-54bp-YFP-C99 and CFP-54bp-YFP were totally same to that base information in gene bank.
2. The purpose fragment was amplified in transfected cells while it was absent in control cells.
3. Cyan and yellow fluorescence could be detected in cells at 12h after transfection. At 24h the intensity of fluorescence strengthened. At 48h the fluorescence increased further. But at 72h the fluorescence became to decrease and by 96h it diminished.
4. FRET occurred in pcDNA3.0-CFP-54bp-YFP transfection cells and the cell shape was clear. FRET was present in pcDNA3.0-CFP-54bp-YFP-C99 transfected-cells at 12h after transfection, but absent thereafter. Much intracellular fluorescence accumulation spread within the cells.
5. In pcDNA3.0-CFP-54bp-YFP transfected-cells, Fret unchanged. In pcDNA3.0-CFP-54bp-YFP-C99 transfected-cells, Fret declined.
6. At 48h in transfection cells, the fusion gene product of CFP-54bp-YFP-C99 could be cleaved byβ- andγ-secretase and liberate Aβ.
7. Aβwas firstly generated within the cell. It accumulated and deposited widespread in the cell and membrane. Very little Aβwas secreted outside of the cell but did not form obvious deposition.
8. The cell shape was abnormal due to Aβgeneration and deposition.
9. Aβwas found by immunocytochemistry.
10. Aβintracellular accumulation resulted in the decrease of transfected-cell viability examined by MTT assay and the difference was significant in comparison with the control group.
Conclusion
1. The construction of recombinant plasmid pcDNA3.0-CFP-54bp-YFP-C99 and pcDNA3.0-CFP-54bp-YFP containing Swedish (and APP717) mutation of APP and two types of fluorescent protein sequences was accomplished.
2. The fusion gene could be expressed correctly at mRNA and protein levels and become to be a strong tool for investigating the mechanism of APP cleavage and Aβgeneration.
3. FRET could be used to detect the occurrence ofβ-cleavage sensitively.
4. CFP-54bp-YFP-C99 could be cleaved byβ-secretase and CFP-54bp-YFP could not which indicated that C99 would be significant forβ-cleavage of APP and might function as signal-peptide for directing the cleavage. If C99 function was disturbed, then the initiation of Aβmight be blocked. It may provide a new idea for the early therapy of AD.
5. The fusion gene could be translated into correct protein. The expression product of fusion gene could be sequentially cleaved byβ- andγ-secretase and generate Aβlabeled by YFP.
6. Aβwas produced mainly and firstly within the cell and little was liberated into the cell space.
7. Before Aβdeposited outside of the cell, Aβaggregated within the cell and induced the secondary damage to the cell which leaded to the cell viability decrease and shape abnormality.
构建含有突变型APP的荧光真核表达系统以研究APP的酶解过程和Aβ产生的分子机制。
方法
以质粒pcDNA3.0-CFP-CaM-YFP的BglII酶切产物为模板,通过聚合酶链式反应(PCR)分别得到编码蓝色荧光蛋白(cyan fluorescence protein, CFP)和黄色荧光蛋白碱基序列(yellow fluorescence protein, YFP);以pcDNA3.0-APP为模板,通过聚合酶链式反应(PCR)得到含有APP717突变的最后300碱基片段(C99);生物合成含有Swedish突变的APP中间54个碱基片段(54bp)。利用基因工程技术将CFP、54bp、YFP、C99片段克隆至载体质粒pcDNA3.0中,得到重组质粒pcDNA3.0-CFP-54bp-YFP-C99和pcDNA3.0-CFP-54bp-YFP,酶切和测序鉴定。将构建的融合基因转染至SH-SY5Y细胞中,通过RT-PCR检测融合基因mRNA表达,利用多光子共聚焦显微镜观察不同时间点(12h、24h、48h、72h、96h)转染细胞内的荧光蛋白表达情况。细胞转染12h、18h、24h、36h、48h、72h后,检测FRET,予以波长为820nm的CFP激发光激发,采集发射波长为473nm的CFP图像,同时采集发射波长为525nm的YFP图像;计算CFP、YFP荧光强度(ICFP和IYFP)和两者的比率(Fret=IYFP/ICFP),绘制曲线。pcDNA3.0-CFP-54bp-YFP-C99转染细胞48h后,利用多光子共聚焦显微镜观察细胞内黄色荧光蛋白的表达、分布以及YFP标记的Aβ去向。免疫细胞化学鉴定Aβ的生成。pcDNA3.0-CFP-54bp-YFP-C99、pcDNA3.0-CFP-54bp-YFP转染细胞12h、24h、36h、48h、72h、96h后,MTT检测转染细胞活性,了解Aβ对细胞的影响。
结果
1、CFP、YFP、C99的PCR产物大小分别为818bp、738bp、325bp,经过不同的双酶切得到的酶切产物大小分别为702bp、723bp和310bp,电泳证实与预期一致;重组质粒pcDNA3.0-CFP-54bp-YFP-C99、pcDNA3.0-CFP-54bp-YFP分别经过不同的组合酶切,得到的目的片段长度正确;测序鉴定证明融合基因CFP-54bp-YFP-C99、CFP-54bp-YFP的序列与“gene bank”的碱基信息相符。
2、将转染细胞进行RT-PCR显示有目的片段,分别对应于CFP-54bp-YFP-C99和YFP-C99,对照细胞内则无相应片段。
3、转染细胞内有蓝色(和)黄色荧光表达,在转染12h后已经可以观察到细胞内有荧光分布,24h荧光逐渐增多,至48h荧光进一步增多增强,72h荧光有所减少,96h荧光明显减少减弱。
4、pcDNA3.0-CFP-54bp-YFP细胞在转染后始终存在FRET,细胞轮廓光滑;pcDNA3.0-CFP-54bp-YFP-C99细胞在转染12h后有微弱FRET,以后无FRET,细胞内有颗粒广泛沉积。
5、CFP-54bp-YFP转染细胞Fret不变; CFP-54bp-YFP-C99转染细胞Fret下降。
6、CFP-54bp-YFP-C99在转染细胞48h后,能够被β、γ分泌酶裂解产生Aβ。
7、Aβ产生于首先细胞内,聚集成颗粒,广泛分布于细胞内和细胞膜上,细胞外间隙也有少量分布,但没有形成明显沉积。
8、Aβ形成后细胞形态异常。
9、免疫细胞化学进一步证实Aβ的生成。
10、MTT证实细胞内聚集的Aβ使得细胞活性下降,与对照组相比差异有统计学意义。
结论
1、荧光标记并含有Swedish (和APP717 )突变的重组质粒pcDNA3.0-CFP-54bp-YFP-C99和pcDNA3.0-CFP-54bp-YFP构建成功。
2、融合基因在mRNA和蛋白水平均能够正确表达,为下一步研究APP裂解和Aβ产生提供了一种有力工具。
3、FRET能够敏感准确的检测APP有无发生β裂解。
4、CFP-54bp-YFP-C99能够被β分泌酶裂解而CFP-54bp-YFP不能被裂解,提示C99可能起到信号肽样的引导定位作用,对β分泌酶裂解APP具有重要意义;干扰C99可能会抑制Aβ的产生,本实验为探索AD的早期干预提供一种新思路。
5、融合基因能够正确表达并被裂解,生成YFP标记的Aβ。
6、Aβ产生于细胞内多个部位,并有少量被分泌至细胞外。
7、Aβ在细胞外形成沉积之前,首先在细胞内聚集并产生继发性细胞毒作用,造成细胞活性下降,形态异常。
Objective
To investigate the mechanism of the APP cleavage progress and Aβgeneration, the construction of recombinant eukaryotic expression plasmid was made encoding Swedish and APP717 mutations of amyloid precursor protein (APP) and fluorescent protein.
Methods
The cyan and yellow fluorescence protein sequences (which were named as CFP and YFP, respectively.) were obtained by polymerase chain reaction (PCR) with the template of BglII-digestion product of the plasmid pcDNA3.0-CFP-CaM-YFP. The last 300 bases of APP sequence (which was named as C99 containing APP717 mutation) were amplified by PCR with the template of the plasmid pcDNA3.0-APP harboring APP717 mutation. The 54 bases in the middle of APP sequence were synthesized (which was named as 54bp containing Swedish mutation) by biological company. The four fragments, mentioned above (which were CFP, YFP, C99 as well as 54bp.) were inserted into the vector pcDNA3.0. By genetic engineering the recombinant plasmid pcDNA3.0-CFP-54bp-YFP-C99 and pcDNA3.0-CFP-54bp-YFP were constructed and identified by enzyme digestion and sequencing assay. The recombinant plasmids pcDNA3.0-CFP-54bp-YFP-C99 and pcDNA3.0-CFP-54bp-YFP were transfected into SH-SY5Y cells respectively. The mRNA expression of the fusion gene was examined by reverse transcript polymerase chain reaction assay (RT-PCR). The protein expression of fusion gene and fluorescence change was detected by multi-photon con-focal microscopy at 12h, 24h, 48h, 72h and 96h after transfection. At 12h, 18h, 24h, 36h, 48h and 72h after transfection of pcDNA3.0-CFP-54bp-YFP-C99 or pcDNA3.0-CFP-54bp-YFP, as soon as the cells were excited at 820nm of CFP excitation wavelength, the CFP images were collected at the CFP emission wavelength of 473nm and YFP images were also collected at the YFP emission wavelength of 525nm. FRET phenomenon was observed to get an understanding ofβ-cleavage of APP. Calculate the intensities of CFP (ICFP) and YFP (IYFP) and then turn them into the ratio Fret ( Fret=IYFP : ICFP). At 48h after pcDNA3.0-CFP-54bp-YFP-C99 transfection, multi-photon con-focal microscope was used to detect the yellow fluorescence, to observe its distribution and trace Aβlabeled by YFP. Aβgeneration was confirmed by immunocytochemistry. The viability of transfection cells was measured via MTT assay to find the Aβeffect on the cells at specific time point.
Results
1. The length of PCR-product of CFP、YFP and C99 was 818bp、738bp and 325bp. The length of product of CFP、YFP and C99 via different double-digestion was 702bp、723bp and 310bp. The electrophoresis result is identical to our expectation. The different double-digestion of the plasmid pcDNA3.0-CFP-54bp-YFP-C99 and pcDNA3.0-CFP-54bp-YFP resulted in different fragments and confirmed the accomplishment of the two recombinant plasmids. Finally sequencing assay showed that the fusion genes of CFP-54bp-YFP-C99 and CFP-54bp-YFP were totally same to that base information in gene bank.
2. The purpose fragment was amplified in transfected cells while it was absent in control cells.
3. Cyan and yellow fluorescence could be detected in cells at 12h after transfection. At 24h the intensity of fluorescence strengthened. At 48h the fluorescence increased further. But at 72h the fluorescence became to decrease and by 96h it diminished.
4. FRET occurred in pcDNA3.0-CFP-54bp-YFP transfection cells and the cell shape was clear. FRET was present in pcDNA3.0-CFP-54bp-YFP-C99 transfected-cells at 12h after transfection, but absent thereafter. Much intracellular fluorescence accumulation spread within the cells.
5. In pcDNA3.0-CFP-54bp-YFP transfected-cells, Fret unchanged. In pcDNA3.0-CFP-54bp-YFP-C99 transfected-cells, Fret declined.
6. At 48h in transfection cells, the fusion gene product of CFP-54bp-YFP-C99 could be cleaved byβ- andγ-secretase and liberate Aβ.
7. Aβwas firstly generated within the cell. It accumulated and deposited widespread in the cell and membrane. Very little Aβwas secreted outside of the cell but did not form obvious deposition.
8. The cell shape was abnormal due to Aβgeneration and deposition.
9. Aβwas found by immunocytochemistry.
10. Aβintracellular accumulation resulted in the decrease of transfected-cell viability examined by MTT assay and the difference was significant in comparison with the control group.
Conclusion
1. The construction of recombinant plasmid pcDNA3.0-CFP-54bp-YFP-C99 and pcDNA3.0-CFP-54bp-YFP containing Swedish (and APP717) mutation of APP and two types of fluorescent protein sequences was accomplished.
2. The fusion gene could be expressed correctly at mRNA and protein levels and become to be a strong tool for investigating the mechanism of APP cleavage and Aβgeneration.
3. FRET could be used to detect the occurrence ofβ-cleavage sensitively.
4. CFP-54bp-YFP-C99 could be cleaved byβ-secretase and CFP-54bp-YFP could not which indicated that C99 would be significant forβ-cleavage of APP and might function as signal-peptide for directing the cleavage. If C99 function was disturbed, then the initiation of Aβmight be blocked. It may provide a new idea for the early therapy of AD.
5. The fusion gene could be translated into correct protein. The expression product of fusion gene could be sequentially cleaved byβ- andγ-secretase and generate Aβlabeled by YFP.
6. Aβwas produced mainly and firstly within the cell and little was liberated into the cell space.
7. Before Aβdeposited outside of the cell, Aβaggregated within the cell and induced the secondary damage to the cell which leaded to the cell viability decrease and shape abnormality.
引文
1. Sukanto S and Lieberburg I. Cellular mechanisms of b-amyloid production and secretion. Proc Natl Acad Sci USA, 1999, 96: 11049–11053.
2. Selkoe D J. Alzheimer’s disease: genes, proteins, and therapy. Physiol Rev , 2001, 81:741–766.
3. Skovronsky DM, Doms RW and Lee VM-Y. Detection of a novel intraneuronal pool of insoluble amyloid b protein that accumulates with time in culture. J Cell Biol 1998,141: 1031–1039.
4. Gouras G, Tsai J, Naslund J, et al. Intraneuronal Ab42 accumulation in human brain. Am J Path, 2000, 156: 15–20.
5. Nunan J, Williamson NA, Hill AF, et al. Proteasome-mediated degradation of the C-terminus of the Alzheimer’s disease beta-amyloid protein precursor: effect of C-terminal truncation on production of beta-amyloid protein. J Neurosci Res, 2003,74:378–385.
6. Haass C, Schlossmacher MG, Hung AY, et al. Amyloid b-peptide is produced by cultured cells during normal metabolism. Nature, 1992, 359: 322–325.
7. Shoji M, Golde TE, Ghiso J, et al. Production of the Alzheimer amyloid b protein by normal proteolytic processing. Science, 1992, 258: 126–129.
8. Esch FS, Keim PS, Beattie EC, et al. Cleavage of amyloid b peptide during constitutive processing of its precursor. Science, 1990, 248: 1122–1124.
9. Haass C, Hung AY, Schlossmacher MG, et al. beta-Amyloid peptide and a 3-kDa fragment are derived by distinct cellular mechanisms. J Biol Chem, 1993, 268: 3021–3024.
10. Knops J, Suomensaari S, Lee M, et al. Cell-type and amyloid precursor protein-type specific inhibition of Abeta release by bafilomycin A1, a selective inhibitor of vacuolar ATPases. J Biol Chem, 1995, 270: 2419–2422.
11. Haass C, Koo EH, Mellon A, et al. Targeting of cell-surface b-amyloid precursor protein to lysosomes: alternative processing into amyloid bearing fragments. Nature 1992, 357: 500–503.
12. Koo EH and Squazzo SL. Evidence that production and release of amyloid beta-protein involves the endocytic pathway. J Biol Chem, 1994, 269: 17386–17389.
13. Haass C, Lemere CA, Capell A, et al. The swedish mutation causes early-onset Alzheimer’s disease by b-secretase cleavage within the secretory pathway. Nat Med 1995, 1: 1291–1296.
14. Xu H, Sweeny D, Wang R, et al. Generation of Alzheimer beta-amyloid protein in the trans-Golgi network in the apparent absence of vescicle formation. Proc Natl Acad Sci USA, 1997, 94: 3748–3752.
15. Cook DG, Forman MS, Sung JC, et al. Alzheimer’s Abeta(1-42) is generated in the endoplasmic reticulum:intermediate compartment of NT2N cells. Nat Med, 1997, 3: 1021–1023.
16. Cook DG, Forman MS, Sung JC, et al. Alzheimer’s A beta(1-42) is generated in the endoplasmic reticulum/intermediate compartment of NT2N cells. Nat Med, 1997, 3: 1021-1023.
17. Wilson CA, Doms RW and Lee VM. Intracellular APPprocessing and A beta production in Alzheimer disease. J Neuropathol Exp Neurol, 1999, 58: 787-794.
18. Huse JT, Pijak DS, Leslie GJ, et al. Maturation and endosomal targeting of beta-site amyloid precursor protein-cleaving enzyme: the Alzheimer’s disease beta-secretase. J Biol Chem, 2000, 275: 33729–33737.
19. Haniu M, Denis P, Young Y, et al. Characterization of Alzheimer’s beta-secretase protein BACE. J Biol Chem, 2000, 275: 21099–21106.
20. Prokop S, Shirotani K, Edbauer D, et al. Requirement of PEN-2 for stabilization of the presenilin N-/Cterminal fragment heterodimer within the gamma-secretase complex. J Biol Chem, 2004, 279: 23 255–23 261.
21. Thinakaran G, Borchelt DR, Lee MK, et al. Endoproteolysis of presenilin 1 and accumulation of processed derivatives in vivo. Neuron, 1996, 17: 181–190.
22. Steiner H, Capell A, Pesold B, et al. Expression of Alzheimer’s disease-associated presenilin-1 is controlled by proteolytic degradation and complex formation. J Biol Chem, 1998, 273: 32322–32331.
23. Annaert W, Levesque L, Craessaerts K, et al. Presenilin 1 controlsγ-secretase processing of amyloid precursor protein in pre-Golgi compartments of hippocampal neurons. J Cell Biol, 1999, 147: 277–294.
24. Xia W, Zhang J, Ostaszewski BL, et al. Presenilin 1 regulates the processing of beta-amyloid precursor protein C-terminal fragments and the generation of amyloid beta-protein in endoplasmic reticulum and Golgi. Biochem, 1998, 37: 16465–16471.
25. Lah JJ and Levey AI. Endogenous presenilin-1 targets to endocytic rather than biosynthetic compartments. Mol Cell Neurosci, 2000, 16: 111–126.
26. Glabe C. Intracellular mechanisms of amyloid accumulation and pathogenesis inAlzheimer’s disease. J Mol Neurosci, 2001, 17: 137-145.
27. Woltjer RL, Nghiem W, Maezawa I, et al. Role of glutathione in intracellular amyloid-a precursor protein/carboxy-terminal fragment aggregation and associated cytotoxicity. J Neurochem, 2005, 93: 1047-1056.
28. Klein WL. A-beta toxicity in Alzheimer’s disease: globular oligomers (ADDLs) as new vaccine and drug targets. Neurochem Intl, 2002, 41: 345–352.
29. Montine T J, Neely M D, Quinn J F, et al. Lipid peroxidation in aging brain and Alzheimer’s disease. Free Radic Biol Med, 2002, 33: 620–626.
30. Dewachter I, Reverse D, Caluwaerts N, et al. Neuronal deficiency of presenilin 1 inhibits amyloid plaque formation and corrects hippocampal long-term potentiation but not a cognitive defect of amyloid precursor protein [V717] transgenic mice. J Neurosci, 2002, 22: 3445-3453.
31. Takahashi RH, Nam EE, Edqar M, et al. Alzheimer beta-amyloid peptides: normal and abnormal localization. Histol Histopathol, 2002, 17: 239-246.
32. Cataldo AM, Petanceska S, Terio NB, et al. Abeta localization in abnormal endosomes: association with earliest Abeta elevations in AD and Down syndrome. Neurobiol Aging, 2004, 25: 1263-1272.
33. Schmitz C, Rutten BP, Pielen A, et al. Hippocampal neuron loss exceeds amyloid plaque load in a transgenic mouse model of Alzheimer’s disease. Am J Pathol, 2004, 164: 1495-1502.
1. Goedert M, Spillantini MG, Cairns NJ, et al. Tau proteins of Alzheimer paired helical filaments : abnormal phosphorylation of all six brain isoforms. Neuron, 1992, 8: 159-168.
2. Lee VM-Y, Balin BJ, Otvos L, et al. A68: a major subunit of paired helical filaments and derivatized forms of normal tau. Science, 1991, 251: 675-678.
3. Probst A, Langui D, Ipsen S, et al. Deposition ofβ/A4 protein along neuronal plasma membranes in diffuse senile plaques. Acta Neuropathol, 1991, 83: 21-29.
4. Lamb BT, Sisodia SS, Lawler AM, et al. Introduction and expression of the 400 kilobases precursor amyloid protein gene in transgenic mice. Nature Genet, 1993, 5: 22-30.
5. Casoli T, Di Stefano G, Giorgetti B, et al. Release of beta-amyloid from high-density platelets: implications for Alzheimer’s disease pathology. Ann N Y Acad Sci, 2007, 1096: 170-178.
6. Monning U, Konig G, Banati RB, et al. Alzheimer beta A4-amyloid protein precursor in immunocompetent cells. J Biol Chem, 1992, 267: 2350-2356.
7. Arai H, Lee VMY, Messinger ML, et al. Expression patterns of beta-amyloid precursor protein in neural and nonneuronal human tissues from Alzheimer’s disease and control subjects. Ann Neurol, 1991, 30: 686-693.
8. Perez R G, Soriano S, Hayes J D, et al. Mutagenesis identified new signals for b-amyloid precursor protein endocytosis, turnover, and the generation of secreted fragments, including Ab42. J Biol Chem, 1999, 274: 18851-18856.
9. Perez R G, Squazzo S L and Koo E H. Enhanced release of amyloid beta-protein from codon 670/671‘Swedish’mutant beta-amyloid precursor protein occurs in both secretory and endocytic pathways. J Biol Chem, 1996, 271: 9100-9107.
10. Ulery P G, Beers J, Mikhailenko I, et al. Modulation of beta-amyloid precursor protein processing by the low density lipoprotein receptor-related protein (LRP). Evidence thatLRP contributes to the pathogenesis of Alzheimer’s disease. J Biol Chem, 2000, 275: 7410-7415.
11. Yamazaki T, Selkoe DJ and Koo EH. Trafficking of cell surface beta-amyloid precursor protein: retrograde and transbytotic transport in cultured neurons. J Cell Biol, 1995, 129: 431-442.
12. Simons M, Ikonen E, Tienari P, et al. Intracellular routing of human amyloid protein precursor axonal delivery followed by transport to the dentrites. J Neurosci Res, 1995, 41: 121-128.
13. Moya KL, Benowitz LI, Scheider GE, et al. The amyloid precursor protein is developmentally regulated and correlated with synaptogenesis. Dev Biol, 1994, 161: 597-603.
14. Nakamura Y, Ikeda Y, Niigawa H, et al. Amyloid beta-protein precursor deposition in rat hippocampus lesioned by ibotenic acid injection. Neurosci. Lett, 1992, 136: 95-98.
15. Otsuka N, Tomanaga M and Ikeda K. Rapid appearance of beta-amyloid precursor protein immunoreactivity in damaged axons and reactive glial cells in rat brain following needle stab injury. Brain Res, 1991, 568: 335-338.
16. Mattson MP, Cheng B, Culwell A, et al. Evidence for excitoprotective and intraneuronal calcium-regulating roles for secreted forms of beta-amyloid precursor protein. Neuron, 1993, 10: 243-254.
17. Allinquant B, Hantraye P, Mailleux P, et al. Downregulation of amyloid precursor protein inhibits neurite outgrowth in vitro. J Cell Biol, 1995, 128: 919-927.
18. Fazeli MS, Breen SK, Errington ML, et al. Increase in extracellular NCAM and amyloid precursor protein following induction of long-term potentiation in the dentate gyrus of anaesthetized rats. Neurosci Lett, 1994, 69: 77-80.
19. Marin S and Baudry M. Properties and mechanism of long term synaptic plasticity in the mammalian brain: relationship to learning and memory. Neurobiol Learn Mem, 1995, 63: 1-18.
20. Smith-Swintosky V, Pettigrew L, Craddock S, et al. Secreted forms of beta-amyloid precursor protein protect against ischemic brain injury. J Neurochem, 1994, 63: 781-784.
21. Jarrett JT, Berger EP, Lansbury PT, et al. The carboxy terminus of the beta- amyloid protein is critical for the seeding of amyloid formation: implications for the pathogenesis of Alzheimer’s disease. Biochemistry, 1993, 32: 4693–4697.
22. Buxbaum JD, Liu KN, Luo Y, et al. Evidence that tumor necrosis factor converting a enzyme is involved in regulated a-secretase cleavage of the Alzheimer amyloid protein precursor. J Biol Chem, 1998, 273: 27765–27767.
23. Skovronsky DM, Moore DB, Milla ME, et al. Protein kinase C-dependent beta-secretase competes with beta-secretase for cleavage of amyloid-beta precursor protein in the trans-Golgi network. J Biol Chem, 2000, 275: 2568–2575.
24. Seubert P, Vigo-Pelfrey C, Esch F, et al. Isolation and quantitation of soluble Alzheimer’s , beta-peptide from biological fluids. Nature, 1992, 359: 325-327.
25. Roberts GW, Gentleman SM, Lynch A, et al. Beta-A4 amyloid protein deposition in brain after head injury. Lancet, 1994, 338: 1422- 1423.
26. Gabuzda D, Busciglio J, Chen L, et al. Inhibition of energy metabolism alters the processing of amyloid precursor protein and induces a potentially amyloidogenie derivative. J Biol Chem, 1994, 269: 13623-13628.
27. Tilley L, Morgan K, Kalsheker N. Genetic risk factors in Alzheimer’s disease. Mol Pathol, 1998, 51: 293–304.
28. Chartier-Harlin MC, Crawford F, Houlden H, et al. Early-onset Alzheimer’s disease caused by mutations at codon 717 of the beta-amyloid precursor protein gene. Nature, 1991, 353: 844–846.
29. Levy E, Carman M D, Fernandez-Madrid I I, et al. Mutation of the Alzheimer’s disease amyloid gene in hereditary cerebral hemorrage, Dutch type. Science, 1990, 248: 1124-1126.
30. Hendricks L, van Duijn C M, Cras P, et al. Presenile dementia and cerebal hemorrhage linked to a mutation at codon 692 of the beta-amyloid precursor protein gene. NatureGenet, 1992, 1:218-221
31. Stein TD, Anders NJ, DeCarli C, et al. Neutralization of transthyretin reverses the neuroprotecitve effects of secreted amyloid precursor protein (APP) in APPsw mice resulting in tau phosphorylation and loss of hippocampal neurons: support for the amyloid hypothesis. J Neurosci, 2004, 24:7707-7717.
32. Asai M, Hattori C, Szab B, et al. Putative function of ADAM9, ADAM10, AND ADAM17 as APP alpha-secretase. Biochem Biophys Res Commu, 2003,301:231-235.
33. Postina R, Schroeder A, Dewachter I, et al. A disintegrin-metalloproteinase prevents amyloid plaque formation and hippocampla defects in an Alzheimer’s disease mouse model. J Clini Invest, 2004, 113: 1456-1464.
34. Ercheberrigaray R, Tan M, Dewachter I, et al. Therapeutic effects of PKC activation in Alzheimer’s disease transgenic mice. Proc Natl Acad Sci USA, 2004, 101: 11141-11146.
35. Vassar R, Bennett BD, Babu-Khan S, et al. Beta-secretase cleavage of Alzheimer’s amyloid precursor protein by the transmembrane aspartic protease BACE. Science, 1999, 286: 735-741
36. Bodendorf U, Danner S, Fischer F, et al. Expression of human beta-secretase in the mouse brain increases the steady-state level of beta-amyloid. J Neurochem, 2002, 80: 799-806.
37. Roberds S L, Anderson J, Basi G, et al. BACE knockout mice are healthy despite lacking the primary beta-secretase activity in brain: implications for Alzheimer’s disease therapeutics. Hum Mol Genet2001, 10: 1317-1324.
38. Sorbi S, Forleo P, Tedde A, et al. Genetic risk factors in familial Alzheimer’s disease. Mech Ageing Dev, 2001, 122(16):1951–60.
39. Beher D, Elle C, Underwood J, et al. Proteolytic fragments of Alzheimer’sdisease-associated presenilin 1 are present in synaptic organelles and growth cone membranes of rat brain. J, Neurochem, 1999, 72:1564–1573.
40. Prokop S, Shirotani K, Edbauer D, et al. Requirement of PEN-2 for stabilization of the presenilin N-/Cterminal fragment heterodimer within the gamma-secretase complex. J Biol Chem, 2004, 279: 23 255–23 261.
41. Ikeuchi T, Sisodia SS. The Notch ligands, Delta1 Jagged2, are substrates for presenilin-dependent“γ-secretase”cleavage. J Biol Chem, 2003,278:7751–4.
42. Murakami D, Okamoto I, Nagano O, et al. Presenilin-dependentγ-secretase activity mediates the intramembranous cleavage of CD44. Oncogene, 2003, 22: 1511–1516.
43. Van Gassen G, Annaert W, Van Broeckhoven C. Binding partners of Alzheimer’s disease proteins: are they physiologically relevant? Neurobiol Dis 2000, 7:135–151.
44. Yuasa S, Nakajima M, Aizawa H, et al. Impaired cell cycle control of neuronal precursor cells in the neocortical primordium of presenilin-1-deficient mice. J Neurosci Res, 2002, 70:501–513.
45. Meredith Jr JE, Wang Q, Mitchell TJ, et al.γ-Secretase activity is not involved in presenilin-mediated regulation ofβ-catenin. Biochem Biophys Res Commun 2002, 299:744–50.
46. Marambaud P, Shioi J, Serban G, et al. A presenilin-1/γ-secretase cleavage releases the E-cadherin intracellular domain and regulates disassembly of adherens junctions. EMBO J, 2002, 21:1948–1956.
47. Leissring MA, LaFerla FM, Callamaras N, et al. Subcellular mechanisms of presenilin-mediated enhancement of calcium signaling. Neurobiol Dis, 2001,8:469–478.
2. Selkoe D J. Alzheimer’s disease: genes, proteins, and therapy. Physiol Rev , 2001, 81:741–766.
3. Skovronsky DM, Doms RW and Lee VM-Y. Detection of a novel intraneuronal pool of insoluble amyloid b protein that accumulates with time in culture. J Cell Biol 1998,141: 1031–1039.
4. Gouras G, Tsai J, Naslund J, et al. Intraneuronal Ab42 accumulation in human brain. Am J Path, 2000, 156: 15–20.
5. Nunan J, Williamson NA, Hill AF, et al. Proteasome-mediated degradation of the C-terminus of the Alzheimer’s disease beta-amyloid protein precursor: effect of C-terminal truncation on production of beta-amyloid protein. J Neurosci Res, 2003,74:378–385.
6. Haass C, Schlossmacher MG, Hung AY, et al. Amyloid b-peptide is produced by cultured cells during normal metabolism. Nature, 1992, 359: 322–325.
7. Shoji M, Golde TE, Ghiso J, et al. Production of the Alzheimer amyloid b protein by normal proteolytic processing. Science, 1992, 258: 126–129.
8. Esch FS, Keim PS, Beattie EC, et al. Cleavage of amyloid b peptide during constitutive processing of its precursor. Science, 1990, 248: 1122–1124.
9. Haass C, Hung AY, Schlossmacher MG, et al. beta-Amyloid peptide and a 3-kDa fragment are derived by distinct cellular mechanisms. J Biol Chem, 1993, 268: 3021–3024.
10. Knops J, Suomensaari S, Lee M, et al. Cell-type and amyloid precursor protein-type specific inhibition of Abeta release by bafilomycin A1, a selective inhibitor of vacuolar ATPases. J Biol Chem, 1995, 270: 2419–2422.
11. Haass C, Koo EH, Mellon A, et al. Targeting of cell-surface b-amyloid precursor protein to lysosomes: alternative processing into amyloid bearing fragments. Nature 1992, 357: 500–503.
12. Koo EH and Squazzo SL. Evidence that production and release of amyloid beta-protein involves the endocytic pathway. J Biol Chem, 1994, 269: 17386–17389.
13. Haass C, Lemere CA, Capell A, et al. The swedish mutation causes early-onset Alzheimer’s disease by b-secretase cleavage within the secretory pathway. Nat Med 1995, 1: 1291–1296.
14. Xu H, Sweeny D, Wang R, et al. Generation of Alzheimer beta-amyloid protein in the trans-Golgi network in the apparent absence of vescicle formation. Proc Natl Acad Sci USA, 1997, 94: 3748–3752.
15. Cook DG, Forman MS, Sung JC, et al. Alzheimer’s Abeta(1-42) is generated in the endoplasmic reticulum:intermediate compartment of NT2N cells. Nat Med, 1997, 3: 1021–1023.
16. Cook DG, Forman MS, Sung JC, et al. Alzheimer’s A beta(1-42) is generated in the endoplasmic reticulum/intermediate compartment of NT2N cells. Nat Med, 1997, 3: 1021-1023.
17. Wilson CA, Doms RW and Lee VM. Intracellular APPprocessing and A beta production in Alzheimer disease. J Neuropathol Exp Neurol, 1999, 58: 787-794.
18. Huse JT, Pijak DS, Leslie GJ, et al. Maturation and endosomal targeting of beta-site amyloid precursor protein-cleaving enzyme: the Alzheimer’s disease beta-secretase. J Biol Chem, 2000, 275: 33729–33737.
19. Haniu M, Denis P, Young Y, et al. Characterization of Alzheimer’s beta-secretase protein BACE. J Biol Chem, 2000, 275: 21099–21106.
20. Prokop S, Shirotani K, Edbauer D, et al. Requirement of PEN-2 for stabilization of the presenilin N-/Cterminal fragment heterodimer within the gamma-secretase complex. J Biol Chem, 2004, 279: 23 255–23 261.
21. Thinakaran G, Borchelt DR, Lee MK, et al. Endoproteolysis of presenilin 1 and accumulation of processed derivatives in vivo. Neuron, 1996, 17: 181–190.
22. Steiner H, Capell A, Pesold B, et al. Expression of Alzheimer’s disease-associated presenilin-1 is controlled by proteolytic degradation and complex formation. J Biol Chem, 1998, 273: 32322–32331.
23. Annaert W, Levesque L, Craessaerts K, et al. Presenilin 1 controlsγ-secretase processing of amyloid precursor protein in pre-Golgi compartments of hippocampal neurons. J Cell Biol, 1999, 147: 277–294.
24. Xia W, Zhang J, Ostaszewski BL, et al. Presenilin 1 regulates the processing of beta-amyloid precursor protein C-terminal fragments and the generation of amyloid beta-protein in endoplasmic reticulum and Golgi. Biochem, 1998, 37: 16465–16471.
25. Lah JJ and Levey AI. Endogenous presenilin-1 targets to endocytic rather than biosynthetic compartments. Mol Cell Neurosci, 2000, 16: 111–126.
26. Glabe C. Intracellular mechanisms of amyloid accumulation and pathogenesis inAlzheimer’s disease. J Mol Neurosci, 2001, 17: 137-145.
27. Woltjer RL, Nghiem W, Maezawa I, et al. Role of glutathione in intracellular amyloid-a precursor protein/carboxy-terminal fragment aggregation and associated cytotoxicity. J Neurochem, 2005, 93: 1047-1056.
28. Klein WL. A-beta toxicity in Alzheimer’s disease: globular oligomers (ADDLs) as new vaccine and drug targets. Neurochem Intl, 2002, 41: 345–352.
29. Montine T J, Neely M D, Quinn J F, et al. Lipid peroxidation in aging brain and Alzheimer’s disease. Free Radic Biol Med, 2002, 33: 620–626.
30. Dewachter I, Reverse D, Caluwaerts N, et al. Neuronal deficiency of presenilin 1 inhibits amyloid plaque formation and corrects hippocampal long-term potentiation but not a cognitive defect of amyloid precursor protein [V717] transgenic mice. J Neurosci, 2002, 22: 3445-3453.
31. Takahashi RH, Nam EE, Edqar M, et al. Alzheimer beta-amyloid peptides: normal and abnormal localization. Histol Histopathol, 2002, 17: 239-246.
32. Cataldo AM, Petanceska S, Terio NB, et al. Abeta localization in abnormal endosomes: association with earliest Abeta elevations in AD and Down syndrome. Neurobiol Aging, 2004, 25: 1263-1272.
33. Schmitz C, Rutten BP, Pielen A, et al. Hippocampal neuron loss exceeds amyloid plaque load in a transgenic mouse model of Alzheimer’s disease. Am J Pathol, 2004, 164: 1495-1502.
1. Goedert M, Spillantini MG, Cairns NJ, et al. Tau proteins of Alzheimer paired helical filaments : abnormal phosphorylation of all six brain isoforms. Neuron, 1992, 8: 159-168.
2. Lee VM-Y, Balin BJ, Otvos L, et al. A68: a major subunit of paired helical filaments and derivatized forms of normal tau. Science, 1991, 251: 675-678.
3. Probst A, Langui D, Ipsen S, et al. Deposition ofβ/A4 protein along neuronal plasma membranes in diffuse senile plaques. Acta Neuropathol, 1991, 83: 21-29.
4. Lamb BT, Sisodia SS, Lawler AM, et al. Introduction and expression of the 400 kilobases precursor amyloid protein gene in transgenic mice. Nature Genet, 1993, 5: 22-30.
5. Casoli T, Di Stefano G, Giorgetti B, et al. Release of beta-amyloid from high-density platelets: implications for Alzheimer’s disease pathology. Ann N Y Acad Sci, 2007, 1096: 170-178.
6. Monning U, Konig G, Banati RB, et al. Alzheimer beta A4-amyloid protein precursor in immunocompetent cells. J Biol Chem, 1992, 267: 2350-2356.
7. Arai H, Lee VMY, Messinger ML, et al. Expression patterns of beta-amyloid precursor protein in neural and nonneuronal human tissues from Alzheimer’s disease and control subjects. Ann Neurol, 1991, 30: 686-693.
8. Perez R G, Soriano S, Hayes J D, et al. Mutagenesis identified new signals for b-amyloid precursor protein endocytosis, turnover, and the generation of secreted fragments, including Ab42. J Biol Chem, 1999, 274: 18851-18856.
9. Perez R G, Squazzo S L and Koo E H. Enhanced release of amyloid beta-protein from codon 670/671‘Swedish’mutant beta-amyloid precursor protein occurs in both secretory and endocytic pathways. J Biol Chem, 1996, 271: 9100-9107.
10. Ulery P G, Beers J, Mikhailenko I, et al. Modulation of beta-amyloid precursor protein processing by the low density lipoprotein receptor-related protein (LRP). Evidence thatLRP contributes to the pathogenesis of Alzheimer’s disease. J Biol Chem, 2000, 275: 7410-7415.
11. Yamazaki T, Selkoe DJ and Koo EH. Trafficking of cell surface beta-amyloid precursor protein: retrograde and transbytotic transport in cultured neurons. J Cell Biol, 1995, 129: 431-442.
12. Simons M, Ikonen E, Tienari P, et al. Intracellular routing of human amyloid protein precursor axonal delivery followed by transport to the dentrites. J Neurosci Res, 1995, 41: 121-128.
13. Moya KL, Benowitz LI, Scheider GE, et al. The amyloid precursor protein is developmentally regulated and correlated with synaptogenesis. Dev Biol, 1994, 161: 597-603.
14. Nakamura Y, Ikeda Y, Niigawa H, et al. Amyloid beta-protein precursor deposition in rat hippocampus lesioned by ibotenic acid injection. Neurosci. Lett, 1992, 136: 95-98.
15. Otsuka N, Tomanaga M and Ikeda K. Rapid appearance of beta-amyloid precursor protein immunoreactivity in damaged axons and reactive glial cells in rat brain following needle stab injury. Brain Res, 1991, 568: 335-338.
16. Mattson MP, Cheng B, Culwell A, et al. Evidence for excitoprotective and intraneuronal calcium-regulating roles for secreted forms of beta-amyloid precursor protein. Neuron, 1993, 10: 243-254.
17. Allinquant B, Hantraye P, Mailleux P, et al. Downregulation of amyloid precursor protein inhibits neurite outgrowth in vitro. J Cell Biol, 1995, 128: 919-927.
18. Fazeli MS, Breen SK, Errington ML, et al. Increase in extracellular NCAM and amyloid precursor protein following induction of long-term potentiation in the dentate gyrus of anaesthetized rats. Neurosci Lett, 1994, 69: 77-80.
19. Marin S and Baudry M. Properties and mechanism of long term synaptic plasticity in the mammalian brain: relationship to learning and memory. Neurobiol Learn Mem, 1995, 63: 1-18.
20. Smith-Swintosky V, Pettigrew L, Craddock S, et al. Secreted forms of beta-amyloid precursor protein protect against ischemic brain injury. J Neurochem, 1994, 63: 781-784.
21. Jarrett JT, Berger EP, Lansbury PT, et al. The carboxy terminus of the beta- amyloid protein is critical for the seeding of amyloid formation: implications for the pathogenesis of Alzheimer’s disease. Biochemistry, 1993, 32: 4693–4697.
22. Buxbaum JD, Liu KN, Luo Y, et al. Evidence that tumor necrosis factor converting a enzyme is involved in regulated a-secretase cleavage of the Alzheimer amyloid protein precursor. J Biol Chem, 1998, 273: 27765–27767.
23. Skovronsky DM, Moore DB, Milla ME, et al. Protein kinase C-dependent beta-secretase competes with beta-secretase for cleavage of amyloid-beta precursor protein in the trans-Golgi network. J Biol Chem, 2000, 275: 2568–2575.
24. Seubert P, Vigo-Pelfrey C, Esch F, et al. Isolation and quantitation of soluble Alzheimer’s , beta-peptide from biological fluids. Nature, 1992, 359: 325-327.
25. Roberts GW, Gentleman SM, Lynch A, et al. Beta-A4 amyloid protein deposition in brain after head injury. Lancet, 1994, 338: 1422- 1423.
26. Gabuzda D, Busciglio J, Chen L, et al. Inhibition of energy metabolism alters the processing of amyloid precursor protein and induces a potentially amyloidogenie derivative. J Biol Chem, 1994, 269: 13623-13628.
27. Tilley L, Morgan K, Kalsheker N. Genetic risk factors in Alzheimer’s disease. Mol Pathol, 1998, 51: 293–304.
28. Chartier-Harlin MC, Crawford F, Houlden H, et al. Early-onset Alzheimer’s disease caused by mutations at codon 717 of the beta-amyloid precursor protein gene. Nature, 1991, 353: 844–846.
29. Levy E, Carman M D, Fernandez-Madrid I I, et al. Mutation of the Alzheimer’s disease amyloid gene in hereditary cerebral hemorrage, Dutch type. Science, 1990, 248: 1124-1126.
30. Hendricks L, van Duijn C M, Cras P, et al. Presenile dementia and cerebal hemorrhage linked to a mutation at codon 692 of the beta-amyloid precursor protein gene. NatureGenet, 1992, 1:218-221
31. Stein TD, Anders NJ, DeCarli C, et al. Neutralization of transthyretin reverses the neuroprotecitve effects of secreted amyloid precursor protein (APP) in APPsw mice resulting in tau phosphorylation and loss of hippocampal neurons: support for the amyloid hypothesis. J Neurosci, 2004, 24:7707-7717.
32. Asai M, Hattori C, Szab B, et al. Putative function of ADAM9, ADAM10, AND ADAM17 as APP alpha-secretase. Biochem Biophys Res Commu, 2003,301:231-235.
33. Postina R, Schroeder A, Dewachter I, et al. A disintegrin-metalloproteinase prevents amyloid plaque formation and hippocampla defects in an Alzheimer’s disease mouse model. J Clini Invest, 2004, 113: 1456-1464.
34. Ercheberrigaray R, Tan M, Dewachter I, et al. Therapeutic effects of PKC activation in Alzheimer’s disease transgenic mice. Proc Natl Acad Sci USA, 2004, 101: 11141-11146.
35. Vassar R, Bennett BD, Babu-Khan S, et al. Beta-secretase cleavage of Alzheimer’s amyloid precursor protein by the transmembrane aspartic protease BACE. Science, 1999, 286: 735-741
36. Bodendorf U, Danner S, Fischer F, et al. Expression of human beta-secretase in the mouse brain increases the steady-state level of beta-amyloid. J Neurochem, 2002, 80: 799-806.
37. Roberds S L, Anderson J, Basi G, et al. BACE knockout mice are healthy despite lacking the primary beta-secretase activity in brain: implications for Alzheimer’s disease therapeutics. Hum Mol Genet2001, 10: 1317-1324.
38. Sorbi S, Forleo P, Tedde A, et al. Genetic risk factors in familial Alzheimer’s disease. Mech Ageing Dev, 2001, 122(16):1951–60.
39. Beher D, Elle C, Underwood J, et al. Proteolytic fragments of Alzheimer’sdisease-associated presenilin 1 are present in synaptic organelles and growth cone membranes of rat brain. J, Neurochem, 1999, 72:1564–1573.
40. Prokop S, Shirotani K, Edbauer D, et al. Requirement of PEN-2 for stabilization of the presenilin N-/Cterminal fragment heterodimer within the gamma-secretase complex. J Biol Chem, 2004, 279: 23 255–23 261.
41. Ikeuchi T, Sisodia SS. The Notch ligands, Delta1 Jagged2, are substrates for presenilin-dependent“γ-secretase”cleavage. J Biol Chem, 2003,278:7751–4.
42. Murakami D, Okamoto I, Nagano O, et al. Presenilin-dependentγ-secretase activity mediates the intramembranous cleavage of CD44. Oncogene, 2003, 22: 1511–1516.
43. Van Gassen G, Annaert W, Van Broeckhoven C. Binding partners of Alzheimer’s disease proteins: are they physiologically relevant? Neurobiol Dis 2000, 7:135–151.
44. Yuasa S, Nakajima M, Aizawa H, et al. Impaired cell cycle control of neuronal precursor cells in the neocortical primordium of presenilin-1-deficient mice. J Neurosci Res, 2002, 70:501–513.
45. Meredith Jr JE, Wang Q, Mitchell TJ, et al.γ-Secretase activity is not involved in presenilin-mediated regulation ofβ-catenin. Biochem Biophys Res Commun 2002, 299:744–50.
46. Marambaud P, Shioi J, Serban G, et al. A presenilin-1/γ-secretase cleavage releases the E-cadherin intracellular domain and regulates disassembly of adherens junctions. EMBO J, 2002, 21:1948–1956.
47. Leissring MA, LaFerla FM, Callamaras N, et al. Subcellular mechanisms of presenilin-mediated enhancement of calcium signaling. Neurobiol Dis, 2001,8:469–478.
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