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Endogenous overproduction of β-amyloid induces tau hyperphosphorylation and decreases the solubility of tau in N2a cells

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Summary.

Although neurofibrillary tangles and senile plaques have been identified as the hallmark pathological changes in the brain of Alzheimer’s disease (AD), the relationship between them is still not fully understood. In the present study, we have studied the effect of endogenously overproduced amyloid β (Aβ) on tau by using wild type amyloid precursor protein (APP) transfected (N2a/APP695), or Swedish mutant APP plus Δ9 deleted presenilin-1 co-transfected (N2a/APPswe.Δ9) and APP vector transfected (N2a/vector) cell lines. We measured the secreted and intracellular Aβ, including Aβ1–40 and Aβ1–42, by Sandwich ELISA assay. It was shown that the levels of Aβ were increased time-dependently in N2a/APP695 and N2a/APPswe.Δ9 but not in N2a/vector upon butyric acid (BA) treatment. Compared with N2a/vector cells, tau in N2a/APP695 and N2a/APPswe.Δ9 cells was not extracted by RIPA buffer, and the SDS-extracted tau protein was hyperphosphorylated at Tau-1 and PHF-1 epitopes upon BA treatment. Obvious accumulation of the hyperphosphorylated tau in N2a/APP695 and N2a/APPswe.Δ9 cells was observed at 48 h after BA treatment. The total level of the extracted tau was reduced in N2a/APP695 and N2a/APPswe.Δ9 lines compared with N2a/vector cells by Western blot, and this reduction of total tau was also detected by immunofluorescence staining. No obvious alteration of tau mRNA was observed in both N2a/APP695 and N2a/APPswe.Δ9 cells compared with N2a/vector. This study provides direct evidence demonstrating that endogenously overproduced Aβ not only induces tau hyperphosphorylation but also decreases the level and solubility of tau in N2a cell lines.

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Abbreviations

AD:

Alzheimer’s disease

FAD:

familial Alzheimer disease

Aβ:

β amyloid

APP:

amyloid precursor protein

PS:

presenilin

BA:

butyric acid

SP:

senile plaque

NFT:

neurofibrillary tangle

BCIP/NBT:

5-bromo-4-chloro-3-indolyl-phosphate/nitroblue tetrazolium

ELISA:

enzyme linked immunosorbent assay

References

  • E Adamec P Mohan JP Vonsattel RA Nixon (2002) ArticleTitleCalpain activation in neurodegenerative diseases: confocal immunofluorescence study with antibodies specifically recognizing the active form of calpain 2 Acta Neuropathol (Berl) 104 92–104 Occurrence Handle1:CAS:528:DC%2BD38XksVaqsr0%3D Occurrence Handle10.1007/s00401-002-0528-6

    Article  CAS  Google Scholar 

  • AD Alonso I Grundke-Iqbal HS Barra K Iqbal (1997) ArticleTitleAbnormal phosphorylation of tau and the mechanism of Alzheimer neurofibrillary degeneration: sequestration of microtubule-associated proteins 1 and 2 and the disassembly of microtubules by the abnormal tau Proc Natl Acad Sci USA 94 298–303 Occurrence Handle8990203 Occurrence Handle1:STN:280:DyaK2s7ksVCgtA%3D%3D Occurrence Handle10.1073/pnas.94.1.298

    Article  PubMed  CAS  Google Scholar 

  • J Avila (2000) ArticleTitleTau aggregation into fibrillar polymers: taupathies FEBS Lett 476 89–92 Occurrence Handle10878257 Occurrence Handle1:CAS:528:DC%2BD3cXktlKqtrw%3D Occurrence Handle10.1016/S0014-5793(00)01676-8

    Article  PubMed  CAS  Google Scholar 

  • L Buee T Bussiere V Buee-Scherrer A Delacourte PR Hof (2000) ArticleTitleTau protein isoforms, phosphorylation and role in neurodegenerative disorders Brain Res Rev 33 95–130 Occurrence Handle10967355 Occurrence Handle1:CAS:528:DC%2BD3cXmtVCrtbY%3D Occurrence Handle10.1016/S0165-0173(00)00019-9

    Article  PubMed  CAS  Google Scholar 

  • J Busciglio A Lorenzo J Yeh BA Yankner (1995) ArticleTitleβ-Amyloid fibrils induce tau phosphorylation and loss of microtubule binding Neuron 14 879–888 Occurrence Handle7718249 Occurrence Handle1:CAS:528:DyaK2MXlt1aiu78%3D Occurrence Handle10.1016/0896-6273(95)90232-5

    Article  PubMed  CAS  Google Scholar 

  • N Canu L Dus C Barbato MT Ciotti C Brancolini AM Rinaldi M Novak A Cattaneo A Bradbury P Calissano (1998) ArticleTitleTau cleavage and dephosphorylation in cerebellar granule neurons undergoing apoptosis J Neurosci 18 7061–7074 Occurrence Handle9736630 Occurrence Handle1:CAS:528:DyaK1cXmt1Sjtrg%3D

    PubMed  CAS  Google Scholar 

  • JA Clemens DT Stephenson (1992) ArticleTitleImplants containing β-amyloid protein are not neurotoxic to young and old rat brain Neurobiol Aging 13 581–586 Occurrence Handle1461346 Occurrence Handle1:CAS:528:DyaK3sXitVekug%3D%3D Occurrence Handle10.1016/0197-4580(92)90059-7

    Article  PubMed  CAS  Google Scholar 

  • DR Davis JP Brion AM Couck JM Gallo DP Hanger K Ladhani C Lewis CCJ Miller T Rupniak C Smith BH Anderton (1995) ArticleTitleThe phosphorylation state of the microtubule-associated protein tau as affected by glutamate, colchicines and β-amyloid in primary rat cortical neuronal cultures Biochem J 309 941–949 Occurrence Handle7639714 Occurrence Handle1:CAS:528:DyaK2MXnt1yisro%3D

    PubMed  CAS  Google Scholar 

  • ZY Fang SJ Liu XC Wang R Liu Q Wang ZY Chen JZ Wang (2003) ArticleTitleEffect of calpain on the degradation of tau protein in rat brain cortex extracts Sheng Wu Hua Xue Yu Sheng Wu Wu Li Xue Bao 35 629–634 Occurrence Handle12883633 Occurrence Handle1:CAS:528:DC%2BD2cXktFymu7s%3D

    PubMed  CAS  Google Scholar 

  • TC Gamblin F Chen A Zambrano A Abraha S Lagalwar AL Guillozet M Lu Y Fu F Garcia-Sierra N LaPointe R Miller RW Berry LI Binder VL Cryns (2003) ArticleTitleCaspase cleavage of tau: linking amyloid and neurofibrillary tangles in Alzheimer’s disease Proc Natl Acad Sci USA 100 10032–10037 Occurrence Handle12888622 Occurrence Handle1:CAS:528:DC%2BD3sXmvVeisrc%3D Occurrence Handle10.1073/pnas.1630428100

    Article  PubMed  CAS  Google Scholar 

  • D Games KM Khan FG Soriano PS Keim DL Davis K Bryant (1992) ArticleTitleLack of Alzheimer pathology after β-amyloid protein injections in rat brain. Neurobiol Aging 13: 569–576 disease Proc Natl Acad Sci USA 100 10032–10037

    Google Scholar 

  • GK Gouras CG Almeida RH Takahashi (2005) ArticleTitleIntraneuronal Aβ accumulation and origin of plaques in Alzheimer’s disease Neurobiol Aging 26 1235–1244 Occurrence Handle16023263 Occurrence Handle1:CAS:528:DC%2BD2MXhtVarsLvE Occurrence Handle10.1016/j.neurobiolaging.2005.05.022

    Article  PubMed  CAS  Google Scholar 

  • J Gotz F Chen J van Dorpe RM Nitsch (2001) ArticleTitleFormation of neurofibrillary tangles in P301l tau transgenic mice induced by Aβ 42 fibrils Science 293 1491–1495 Occurrence Handle11520988 Occurrence Handle1:STN:280:DC%2BD3MvnslKjsA%3D%3D Occurrence Handle10.1126/science.1062097

    Article  PubMed  CAS  Google Scholar 

  • I Grundke-Iqbal K Iqbal M Quinlan YC Tung MS Zaidi HM Wisniewski (1986) ArticleTitleMicrotubule-associated protein tau: A component of Alzheimer paired helical filaments J Biol Chem 261 6084–6089 Occurrence Handle3084478 Occurrence Handle1:CAS:528:DyaL28XitVCqsb0%3D

    PubMed  CAS  Google Scholar 

  • P Han F Dou F Li X Zhang YW Zhang H Zheng SA Lipton H Xu FF Liao (2005) ArticleTitleSuppression of cyclin-dependent kinase 5 activation by amyloid precursor protein: A novel excitoprotective mechanism involving modulation of Tau phosphorylation J Neurosci 25 11542–11552 Occurrence Handle16354912 Occurrence Handle1:CAS:528:DC%2BD28XhsVektg%3D%3D Occurrence Handle10.1523/JNEUROSCI.3831-05.2005

    Article  PubMed  CAS  Google Scholar 

  • J Hardy DJ Selkoe (2002) ArticleTitleThe amyloid hypothesis of Alzheimer’s disease: progress and problems on the road to therapeutics Science 297 353–356 Occurrence Handle12130773 Occurrence Handle1:CAS:528:DC%2BD38Xls1Cju7s%3D Occurrence Handle10.1126/science.1072994

    Article  PubMed  CAS  Google Scholar 

  • T Harkany I Abraham C Konya C Nyakas M Zarandi B Penke PG Luiten (2000a) ArticleTitleMechanisms of β-amyloid neurotoxicity: perspectives of pharmacotherapy Rev Neurosci 11 329–382 Occurrence Handle1:STN:280:DC%2BD3crhtVygtw%3D%3D

    CAS  Google Scholar 

  • T Harkany I Abraham W Timmerman G Laskay B Toth M Sasvari C Konya JB Sebens J Korf C Nyakas M Zarandi K Soos B Penke PG Luiten (2000b) ArticleTitleβ-Amyloid neurotoxicity is mediated by a glutamate-triggered excitotoxic cascade in rat nucleus basalis Eur J Neurosci 12 2735–2745 Occurrence Handle1:STN:280:DC%2BD3cvptlahtw%3D%3D Occurrence Handle10.1046/j.1460-9568.2000.00164.x

    Article  CAS  Google Scholar 

  • M Hoshi M Sato S Matsumoto A Noguchi K Yasutake N Yoshida K Sato (2003) ArticleTitleSpherical aggregates of β-amyloid (amylospheroid) show high neurotoxicity and activate tau protein kinase I/glycogen synthase kinase-3β Proc Natl Acad Sci USA 100 6370–6375 Occurrence Handle12750461 Occurrence Handle1:CAS:528:DC%2BD3sXktlyhtLo%3D Occurrence Handle10.1073/pnas.1237107100

    Article  PubMed  CAS  Google Scholar 

  • K Iqbal I Grundke-Iqbal AJ Smith L George YC Tung T Zaidi (1989) ArticleTitleIdentification and localization of a tau peptide to paired helical filaments of Alzheimer disease Proc Natl Acad Sci USA 86 5646–5650 Occurrence Handle2501795 Occurrence Handle1:CAS:528:DyaL1MXlslWqs7g%3D Occurrence Handle10.1073/pnas.86.14.5646

    Article  PubMed  CAS  Google Scholar 

  • MC Irizarry M McNamara K Fedorchak K Hsiao BT Hyman (1997a) ArticleTitleAPPSw transgenic mice develop age-related Aβ deposits and neuropil abnormalities, but no neuronal loss in CA1 J Neuropathol Exp Neurol 56 965–973 Occurrence Handle1:CAS:528:DyaK2sXmtVKqtb0%3D

    CAS  Google Scholar 

  • MC Irizarry F Soriano M McNamara KJ Page D Schenk D Games BT Hyman (1997b) ArticleTitleAβ deposition is associated with neuropil changes, but not with overt neuronal loss in the human amyloid precursor protein V717F (PDAPP) transgenic mouse J Neurosci 17 7053–7059 Occurrence Handle1:CAS:528:DyaK2sXmtVKmurg%3D

    CAS  Google Scholar 

  • Y Ito M Ito N Takagi H Saito K Ishige (2003) ArticleTitleNeurotoxicity induced by amyloid β-peptide and ibotenic acid in organotypic hippocampal cultures: protection by S-allyl-L-cysteine, a garlic compound Brain Res 985 98–107 Occurrence Handle12957372 Occurrence Handle1:CAS:528:DC%2BD3sXmvVyjtrc%3D Occurrence Handle10.1016/S0006-8993(03)03173-1

    Article  PubMed  CAS  Google Scholar 

  • T Iwatsubo A Odaka N Suzuki H Mizusawa N Nukina Y Ihara (1994) ArticleTitleVisualization of Aβ42(43) and Aβ40 in senile plaques with end-specific Aβ monoclonals: evidence that an initially deposited species is Aβ42(43) Neuron 13 45–53 Occurrence Handle8043280 Occurrence Handle1:CAS:528:DyaK2cXlsFCkt74%3D Occurrence Handle10.1016/0896-6273(94)90458-8

    Article  PubMed  CAS  Google Scholar 

  • GV Johnson RS Jope LI Binder (1989) ArticleTitleProteolysis of tau by calpain Biochem Biophys Res Commun 163 1505–1511 Occurrence Handle2551295 Occurrence Handle1:CAS:528:DyaL1MXmt1Ontb0%3D Occurrence Handle10.1016/0006-291X(89)91150-9

    Article  PubMed  CAS  Google Scholar 

  • A Kampfl JS Whitson X Zhao R Posmantur GL Clifton RL Hayes (1995) ArticleTitleCalpain inhibitors reduce depolarization induced loss of tau protein in primary septo-hippocampal cultures Neurosci Lett 194 149–152 Occurrence Handle7478225 Occurrence Handle1:CAS:528:DyaK2MXntl2mtb0%3D Occurrence Handle10.1016/0304-3940(95)11745-I

    Article  PubMed  CAS  Google Scholar 

  • MP Lambert G Stevens S Sabo K Barber G Wang W Wade G Krafft S Snyder TF Holzman WL Klein (1994) ArticleTitleβ/A4-evoked degeneration of differentiated SH-SY5Y human neuroblastoma cells J Neurosci Res 39 377–385 Occurrence Handle7533843 Occurrence Handle1:CAS:528:DyaK2cXmvFyiur4%3D Occurrence Handle10.1002/jnr.490390404

    Article  PubMed  CAS  Google Scholar 

  • W Le WJ Xie R Kong SH Appel (1997) ArticleTitleβ-Amyloid induced neurotoxicity of a hybrid septal cell line associated with increased tau phosphorylation and expression of β-amyloid precursor protein J Neurochem 69 978–985 Occurrence Handle9282919 Occurrence Handle1:CAS:528:DyaK2sXls1Cht78%3D Occurrence Handle10.1046/j.1471-4159.1997.69030978.x

    Article  PubMed  CAS  Google Scholar 

  • J Lewis DW Dickson WL Lin L Chisholm A Corral G Jones SH Yen N Sahara L Skipper D Yager C Eckman J Hardy M Hutton E McGowan (2001) ArticleTitleEnhanced neurofibrillary degeneration in transgenic mice expressing mutant tau and APP Science 293 1487–1491 Occurrence Handle11520987 Occurrence Handle1:CAS:528:DC%2BD3MXmsVeru7Y%3D Occurrence Handle10.1126/science.1058189

    Article  PubMed  CAS  Google Scholar 

  • L Pennanen J Gotz (2005) ArticleTitleDifferent tau epitopes define Aβ42-mediated tau insolubility Biochem Biophys Res Commun 337 1097–1101 Occurrence Handle16226718 Occurrence Handle1:CAS:528:DC%2BD2MXhtFKhs7nJ Occurrence Handle10.1016/j.bbrc.2005.09.168

    Article  PubMed  CAS  Google Scholar 

  • L Qin Y Liu C Cooper B Liu B Wilson JS Hong (2002) ArticleTitleMicroglia enhance β-amyloid peptide-induced toxicity in cortical and mesencephalic neurons by producing reactive oxygen species J Neurochem 83 973–983 Occurrence Handle12421370 Occurrence Handle1:CAS:528:DC%2BD38XovFOisbs%3D Occurrence Handle10.1046/j.1471-4159.2002.01210.x

    Article  PubMed  CAS  Google Scholar 

  • M Ramsden Z Henderson HA Pearson (2002) ArticleTitleModulation of Ca2+ channel currents in primary cultures of rat cortical neurones by amyloid β protein (1–40) is dependent on solubility status Brain Res 956 254–261 Occurrence Handle12445693 Occurrence Handle1:CAS:528:DC%2BD38Xoslamsb4%3D Occurrence Handle10.1016/S0006-8993(02)03547-3

    Article  PubMed  CAS  Google Scholar 

  • JS Richardson Y Zhou U Kumar (1996) ArticleTitleFree radicals in the neurotoxic actions of β-amyloid Ann N Y Acad Sci 777 362–367 Occurrence Handle8624113 Occurrence Handle1:CAS:528:DyaK28XjtVClu7k%3D

    PubMed  CAS  Google Scholar 

  • A Roher D Wolfe M Palutke D KuKuruga (1986) ArticleTitlePurification, ultrastructure, and chemical analysis of Alzheimer disease amyloid plaque core protein Proc Natl Acad Sci USA 83 2662–2666 Occurrence Handle3458224 Occurrence Handle1:CAS:528:DyaL28Xit1art7Y%3D Occurrence Handle10.1073/pnas.83.8.2662

    Article  PubMed  CAS  Google Scholar 

  • DJ Selkoe (1996) ArticleTitleAmyloid β-protein and the genetics of Alzheimer’s disease J Biol Chem 271 18295–18298 Occurrence Handle8756120 Occurrence Handle1:CAS:528:DyaK28XkslCjtb8%3D

    PubMed  CAS  Google Scholar 

  • A Takeuchi MC Irizarry K Duff TC Saido K Hsiao Ashe M Hasegawa DM Mann BT Hyman T Iwatsubo (2000) ArticleTitleAge-related amyloid β deposition in transgenic mice overexpressing both Alzheimer mutant presenilin 1 and amyloid β precursor protein Swedish mutant is not associated with global neuronal loss Am J Pathol 157 331–339 Occurrence Handle10880403 Occurrence Handle1:CAS:528:DC%2BD3cXlsVGmtLk%3D

    PubMed  CAS  Google Scholar 

  • AK Tickler JD Wade F Separovic (2005) ArticleTitleThe role of Aβ peptides in Alzheimer’s disease Protein Pept Lett 12 513–519 Occurrence Handle16101387 Occurrence Handle1:CAS:528:DC%2BD2MXmvV2hur8%3D Occurrence Handle10.2174/0929866054395905

    Article  PubMed  CAS  Google Scholar 

  • RS Turner N Suzuki AS Chyung SG Younkin VM Lee (1996) ArticleTitleAmyloids β40 and β42 are generated intracellularly in cultured human neurons and their secretion increases with maturation J Biol Chem 271 8966–8970 Occurrence Handle8621541 Occurrence Handle1:CAS:528:DyaK28Xitlent7c%3D Occurrence Handle10.1074/jbc.271.15.8966

    Article  PubMed  CAS  Google Scholar 

  • A Wu CA Derrico L Hatem RA Colvin (1997) ArticleTitleAlzheimer’s amyloid-β peptide inhibits sodium/calcium exchange measured in rat and human brain plasma membrane vesicles Neuroscience 80 675–684 Occurrence Handle9276485 Occurrence Handle1:CAS:528:DyaK2sXls1Olur4%3D Occurrence Handle10.1016/S0306-4522(97)00053-5

    Article  PubMed  CAS  Google Scholar 

  • YC Zhang ZF Wang Q Wang YP Wang JZ Wang (2004) ArticleTitleMelatonin attenuates overproduction of β-amyloid-induced inhibition in expression of neurofilament protein Acta Pharmacol Sin 25 447–451 Occurrence Handle15066211 Occurrence Handle1:CAS:528:DC%2BD2cXjtFKgsLs%3D

    PubMed  CAS  Google Scholar 

  • WH Zheng S Bastianetto F Mennicken W Ma S Kar (2002) ArticleTitleAmyloid β peptide induces tau phosphorylation and loss of cholinergic neurons in rat primary septal cultures Neuroscience 115 201–211 Occurrence Handle12401334 Occurrence Handle1:CAS:528:DC%2BD38XotVWhtb0%3D Occurrence Handle10.1016/S0306-4522(02)00404-9

    Article  PubMed  CAS  Google Scholar 

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Authors and Affiliations

  1. Pathophysiology Department, Huazhong University of Science and Technical, Wuhan, China

    Y.-P. Wang, X.-C. Wang, Q. Tian, Y. Yang, Q. Zhang, J.-Y. Zhang, Y.-C. Zhang, Z.-F. Wang, Q. Wang & J.-Z. Wang

  2. Hubei Provincial Key Laboratory in Neurological Diseases, Tongji Medical College, Huazhong University of Science and Technical, Wuhan, China

    X.-C. Wang, Q. Tian, Y. Yang, J.-Y. Zhang, Q. Wang, H. Li & J.-Z. Wang

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  1. Y.-P. Wang
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  2. X.-C. Wang
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  3. Q. Tian
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  4. Y. Yang
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  5. Q. Zhang
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  9. Q. Wang
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  10. H. Li
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  11. J.-Z. Wang
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The first and second authors contributed equally to the paper

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Wang, YP., Wang, XC., Tian, Q. et al. Endogenous overproduction of β-amyloid induces tau hyperphosphorylation and decreases the solubility of tau in N2a cells. J Neural Transm 113, 1723–1732 (2006). https://doi.org/10.1007/s00702-006-0507-5

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