Reactive microglia drive tau pathology and contribute to the spreading of pathological tau in the brain

Brain, Journal Year: 2015, Volume and Issue: 138(6), P. 1738 - 1755

Published: March 31, 2015

Pathological aggregation of tau is a hallmark Alzheimer's disease and related tauopathies. We have previously shown that the deficiency microglial fractalkine receptor (CX3CR1) led to acceleration pathology memory impairment in an hTau mouse model tauopathy. Here, we show microglia drive cell-autonomous manner. First, hyperphosphorylation occur as early 2 months age hTauCx3cr1−/− mice. Second, CD45+ activation correlates with spatial deficit spread anatomically connected regions hippocampus. Third, adoptive transfer purified derived from mice induces within brains non-transgenic recipient Finally, inclusion interleukin 1 antagonist (Kineret®) inoculum significantly reduces microglia-induced pathology. Together, our results suggest reactive are sufficient correlate pathological brain. Neuroinflammation accelerates pathology, but role played by uncertain. Maphis et al. provide direct evidence protein manner,

Language: Английский

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Physiologic brain activity causes DNA double-strand breaks in neurons, with exacerbation by amyloid-β

Nature Neuroscience, Journal Year: 2013, Volume and Issue: 16(5), P. 613 - 621

Published: March 22, 2013

Language: Английский

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Tau post-translational modifications in wild-type and human amyloid precursor protein transgenic mice

Nature Neuroscience, Journal Year: 2015, Volume and Issue: 18(8), P. 1183 - 1189

Published: July 20, 2015

Language: Английский

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The Synaptic Accumulation of Hyperphosphorylated Tau Oligomers in Alzheimer Disease Is Associated With Dysfunction of the Ubiquitin-Proteasome System

American Journal Of Pathology, Journal Year: 2012, Volume and Issue: 181(4), P. 1426 - 1435

Published: Aug. 4, 2012

In Alzheimer disease (AD), deposition of neurofibrillary tangles and loss synapses in the neocortex limbic system each correlate strongly with cognitive impairment. Tangles are composed misfolded hyperphosphorylated tau proteins; however, link between abnormalities synaptic dysfunction remains unclear. We examined location control AD cortices using biochemical morphologic methods. found that, addition to its well-described axonal localization, normal is present at both presynaptic postsynaptic terminals human brains. AD, becomes terminals, this abnormally posttranslationally modified enriched synaptoneurosomal fractions. Synaptic seems be ubiquitinated, forms stable oligomers resistant SDS denaturation. The accumulation associated increased ubiquitinated substrates proteasome components, consistent ubiquitin-proteasome system. Our findings suggest that may an important mediator proteotoxicity disrupts AD. (AD) most common neurodegenerative disorder elderly, affects primarily system, complex pathophysiologic features include inclusions (neurofibrillary tangles, neuropil threads, dystrophic neurites), β-amyloid (plaques cerebral amyloid angiopathy), neurons synapses, astrogliosis, microglial activation, inflammation.1Serrano-Pozo A. Frosch M.P. Masliah E. Hyman B.T. Neuropathological alterations disease.Cold Spring Harb Perspect Med. 2011; 1: a006189Crossref PubMed Scopus (2195) Google Scholar, 2Braak Griffing K. Arai Bohl J. Bratzke H. Braak Neuropathology Alzheimer's disease: what new since Alzheimer?.Eur Arch Psychiatry Clin Neurosci. 1999; 249: 14-22Crossref Scholar Among these features, loss3Terry R.D. Salmon D.P. Butters N. DeTeresa R. Hill Hansen L.A. Katzman Physical basis synapse major impairment.Ann Neurol. 1991; 30: 572-580Crossref (3511) 4DeKosky S.T. Scheff S.W. Synapse frontal cortex biopsies correlation severity.Ann 1990; 27: 457-464Crossref (1764) tangle deposition5Arriagada P.V. Marzloff Distribution Alzheimer-type pathologic changes nondemented elderly individuals matches pattern disease.Neurology. 1992; 42: 1681-1688Crossref 6Giannakopoulos P. Herrmann F.R. Bussiere T. Bouras C. Kovari Perl Morrison J.H. Gold G. Hof P.R. Tangle neuron numbers, but not load, predict status 2003; 60: 1495-1500Crossref (828) seem best decline. Neurofibrillary correlated clinicopathologic studies AD7Ingelsson M. Fukumoto Newell K.L. Growdon Hedley-Whyte E.T. Albert M.S. Irizarry M.C. Early Abeta progressive loss, gliosis, formation brain.Neurology. 2004; 62: 925-931Crossref (542) Scholar; whether a co-occurrence parallel processes or more directly related biology uncertain.8Morris Maeda S. Vossel Mucke L. many faces tau.Neuron. 70: 410-426Abstract Full Text PDF (674) 9Spires-Jones T.L. Stoothoff W.H. de Calignon Jones P.B. Tau pathophysiology neurodegeneration: tangled issue.Trends 2009; 32: 150-159Abstract (281) We, therefore, used techniques address issue protein adult brain Normal abundant microtubule-associated has been described as predominantly localized axons mature neurons.10Dotti C.G. Banker G.A. Binder L.I. expression distribution proteins 2 hippocampal rat situ cell culture.Neuroscience. 1987; 23: 121-130Abstract (173) folded (p-tau) accumulates axons, dendrites, somas.11Avila Lucas J.J. Perez Hernandez F. Role physiological pathological conditions.Physiol Rev. 84: 361-384Crossref (721) 12Brandt Hundelt Shahani alteration neuronal degeneration tauopathies: mechanisms models.Biochim Biophys Acta. 2005; 1739: 331-354Crossref (205) contrast long-held generalizations, recent reports have suggested also normally dendritic spines, where it interacts density (PSD) such Fyn kinase.13Ittner L.M. Ke Y.D. Delerue Bi Gladbach van Eersel Wolfing Chieng B.C. Christie M.J. Napier I.A. Eckert Staufenbiel Hardeman Gotz Dendritic function mediates amyloid-beta toxicity mouse models.Cell. 2010; 142: 387-397Abstract (1418) hypothesized pathologically accumulate sites because recently can locales mice,13Ittner AD,14Cripps D. Thomas S.N. Jeng Y. Yang Davies A.J. disease-specific conformation paired helical filament-Tau polyubiquitinated through Lys-48, Lys-11, Lys-6 ubiquitin conjugation.J Biol Chem. 2006; 281: 10825-10838Abstract (227) 15Morishima-Kawashima Hasegawa Takio Suzuki Titani Ihara Ubiquitin conjugated amino-terminally processed filaments.Neuron. 1993; 10: 1151-1160Abstract (302) site ubiquitination proteasome-mediated degradation structures.16Bingol B. Schuman E.M. Activity-dependent dynamics sequestration proteasomes spines.Nature. 441: 1144-1148Crossref (275) 17Yi Ehlers M.D. Emerging roles for function.Pharmacol 2007; 59: 14-39Crossref (177) By isolating we observed brains, terminals. contrast, synaptoneurosomes isolated from brains p-tau form SDS-resistant on sides synapse, showing enrichment when compared cytoplasm. mirrors same fraction, well chaperones, which suggests aggregates impaired proteolysis mediated by (UPS).18Keck Nitsch Grune Ullrich O. Proteasome inhibition filament-tau patients disease.J Neurochem. 85: 115-122Crossref (405) Protease inhibitor (cOmplete tablet) was purchased Roche Applied Science (Roche Diagnostics Corp., Indianapolis, IN). Phosphatase cocktails 3 were Sigma-Aldrich Corp. (St. Louis, MO) 1:1 combination. Mouse monoclonal antibodies PHF1 (pS396/pS404 tau), CP13 (pS202 DA9 (total tau) gifts Peter (Albert Einstein College Medicine, Bronx, NY). Rabbit anti–total (A20024) Dako Denmark A/S (Glostrup, Denmark); rabbit anti-PSD95 (No. 2507) Cell Signaling Technology, Inc. (Danvers, MA); anti-MBP (SMI-99P) Covance, (Princeton, NJ); anti-actin (A4700), (A5060), anti-tubulin β3 (T8660) Sigma-Aldrich; anti-synaptophysin (AB8049), anti-VCP (AB19444), anti-VDAC (AB34726) Abcam (Cambridge, anti-GAPDH (MAB374), anti-histone H3 (05-928), anti-Myc (06-549) Millipore (Billerica, anti-ubiquitin conjugates (UG9510), anti-α7 (20S subunit, PW8110), anti-Rpt1 (26S PW8852) Enzo Clinical Laboratories, (Farmingdale, Brains subjects diagnosis no deficits obtained Massachusetts Disease Research Center General Hospital Department. All donor tissue accord local national institutional review board regulations. Characteristics controls quantitative analyses given Table 1.Table 1Characteristics Control AD-Affected Used Quantitative StudiesCase No.Age (years)SexClinical DiagnosisDisease Duration (years)ApoE GenotypePMI (hours)Braak StageExperimentC189FControlNA2/3132Figure 4C291FControlNA3/3192Figure 4C371MControlNANA50Figure 4C487MControlNANA361Figure 4C580FControlNA2/4541Figure 7C676MControlNA3/4481Figure 7C785MControlNA3/3242Figure 7C857FControlNA3/3130Figure 7C974FControlNA3/3241Figure 7C1088FControlNA3/3202Figure 7AD183FAD133/4125Figure 4AD282MAD63/476Figure 4AD391FAD143/495Figure 4AD495MADNA3/3116Figure 4AD585FAD43/4105Figure 5AD673FAD193/3145Figure 5AD784FAD163/4125Figure 5AD865MAD83/3215Figures 7, 8AD984FAD73/376Figures 8AD1075FAD53/3266Figures 8AD1192FAD93/346Figures 8AD1293MAD173/365Figures 8AD1374MAD113/3155Figures 8AD1492MAD224/4125Figures 5, 8AD1568FAD114/4206Figures 8AD1680FAD124/4116Figures 8AD1771FAD174/4146Figures 8AD1874MAD174/4205Figures 8AD1989MAD104/4105Figures 8F = female; M male; disease; NA applicable available; PMI postmortem interval. Open table tab F Cortical gray matter (200 300 mg) taken frozen gently ground Potter-Elvehjem homogenizer 1.5 mL ice-cold buffer A (25 mmol/L HEPES [pH 7.5], 120 NaCl, 5 KCl, 1 MgCl2, CaCl2) supplemented dithiothreitol (DTT), protease inhibitors, phosphatase inhibitors. homogenate passed two layers 80-μm nylon filters (Millipore) remove debris, 200-μL aliquot saved. saved mixed 200 μL water 70 10% SDS, 27-gauge needle, boiled minutes prepare total extract. To filtered Hollingsworth et al,19Hollingsworth E.B. McNeal Burton J.L. Williams R.J. Daly J.W. Creveling C.R. Biochemical characterization synaptoneurosome preparation guinea pig cortex: cyclic adenosine 3′:5′-monophosphate-generating systems, receptors, enzymes.J 1985; 5: 2240-2253Crossref remainder 5-μm Supor membrane filter (Pall Port Washington, NY) large organelles nuclei, centrifuged 1000 × g minutes. pellet washed once again, yielding pellet. Supernatant first centrifugation clarified via 100,000 hour obtain cytosol fraction. Cytosolic extract prepared adding 1.5% boiling Synaptoneurosome pellets extracted 0.5 B (50 Tris DTT), Flotation sucrose gradient previously published procedures.20Lathia Wesemann W. Serotonin uptake release biochemically characterized nerve endings concomitant flotation sedimentation centrifugation.J Neural Transm. 1975; 37: 111-126Crossref (4) Ice-cold solutions 0.3, 0.93, 1.2 mol/L 10 (pH 7.5). resuspended transferred centrifuge tube, then overlaid 1.2, 0.3 three-layer discontinuous gradient. After 60,000 hours, collected 0.93/1.2-mol/L interface, myelin 0.3/0.93-mol/L diluted about final concentration. sucrose. three fractions 20,000 20 minutes, C [50 7.5), 150 2% Triton X-100, inhibitors]. incubation rotating mixture hour. supernatant 1% SDS-denatured extracts 15 followed bicinchoninic acid assays (Pierce Protein Biology, Fisher Scientific, Inc., Rockford, IL) determine concentrations. Extracts again after 5X sample (250 5% 400 DTT, 50% glycerol, 0.2% Orange G). Samples resolved SDS-PAGE Bis-Tris 4% 12% gels (Invitrogen Carlsbad, CA), low-fluorescence polyvinylidene difluoride nitrocellulose membranes immunoblotting, detected Odyssey laser scanner (LI-COR Biosciences, Lincoln, NE). Blocking buffer, stripping secondary LI-COR, according manufacturer's protocols. D (20 4 cleared 15,000 equal volume E sodium deoxycholate) neutralize SDS. pre-cleared G Sepharose (GE Healthcare, Pittsburg, PA) 4°C hour, overnight S5a UIM agarose conjugate (UW9820; Laboratories) sepharose (control). flow-through resin times cold X-100) 1.5X elute captured proteins. immunoprecipitation experiments, incubated (control), hours. Western blots enhanced chemiluminescence TrueBlot Ultra anti-mouse horseradish peroxidase (eBioscience, San Diego, CA) Clean-Blot (against primary antibodies; Pierce) minimize signals denatured IgG. A, needles, paraformaldehyde PBS-MC (1 MgCl2 Lab-Tek II CC2 pre-coated chamber slides (Nunc, Rochester, settling 4°C, became fixed attached glass surface, (room temperature point on). Synaptoneurosomes permeabilized 0.05% X-100 3% bovine serum albumen (BSA), times. Slides blocked goat BSA 30 washes. Secondary slide mounted No. coverslip Prolong Antifade reagent (Invitrogen). Primary immunostaining included anti-vGlut1 (Millipore AB590, 1:150), chicken anti-MAP2 (Abcam AB5392, 100), AB12093, 1:100), (1:150), (1:80). Fluorescent donkey Jackson ImmunoResearch (West Grove, 1:100 dilutions (anti-guinea DyLight 649, anti-chicken Cy3, anti-goat Alexa 488, 488). brightfield images immunostained acquired Axio Imager Z epifluorescence microscope (Carl Zeiss AG, Oberkochen, Germany) equipped 63X oil immersion objective (numerical aperture, 1.40). Images deconvolved Iterative Deconvolution plug-in (by Bob Dougherty, OptiNav, Inc.) ImageJ software (version 1.44). co-localization determined manual analysis randomly selected areas wide-field images. glutaraldehyde PBS rinsed, post-fixed osmium tetroxide, embedded LR White (Electron Microscopy Sciences, Hatfield, PA). transmission electron ATM digital camera (JEOL1011; JEOL USA, Peabody, MA). quantified densitometry gel Statistical tests (paired t-test, Mann-Whitney test, two-way variance, linear regression) performed statistical (GraphPad version 5.03; Prism Software La Jolla, CA). Proteins cortical homogenates analyzed SDS-PAGE. extracts, exhibited only monomeric nonhyperphosphorylated 65 kDa). AD-affected contained species (reactive against CP1321Duff Knight Refolo Sanders Yu X. Picciano Malester Hutton Adamson Goedert Burki Characterization pathology transgenic mice over-expressing genomic cDNA transgenes.Neurobiol Dis. 2000; 7: 87-98Crossref (251) PHF122Greenberg S.G. Schein J.D. Hydrofluoric acid-treated PHF display properties tau.J 267: 564-569Abstract Scholar) migrated smear (Figure 1). Fast-migrating (15 50 kDa) represented truncated p-tau; slow-migrating (65 several hundred oligomers,23Watanabe Deamidation isoaspartate smeared filaments Unusual microtubule-binding domain 274: 7368-7378Abstract (106) stably denaturation reducing agents. understand subcellular localization separated into cytosolic (synaptoneurosome) under microscopy, showed well-preserved structures. synaptoneurosomes, fibrillar microscopy 2A). samples nonphosphorylated than expected revealed accumulated high levels remained surprisingly absent 2B). Considered together, our data nonfibrillar pellet, containing oligomers. examine rather originating other contaminants inclusions, further purify 3A). One potential concern could co-precipitated during low-speed (1000 g). Because assemblies dense lipid-rich vesicles, chose gradient–purified great reductions organelle nuclei 3B). they low clearly detectable tau, suggesting confirming role synapse.13Ittner 24Roberson E.D. Scearce-Levie Palop Yan Cheng I.H. Wu Gerstein G.Q. Reducing endogenous ameliorates beta-induced model.Science. 316: 750-754Crossref (1545) including aggregate 3C), hyperphosphorylation oligomerization specific synapses. did demonstrate terminals; developed protocol this. visualize spread dilute solution onto lightly place (see Materials Methods). vesicular glutamate transporter (vGlut1) marker, (MAP2) marker 4A). MAP2 lower inside spines PSD.25Morales Fifkova colocalization actin: immunogold electron-microscope study.Cell Tissue Res. 1989; 256: 447-456Crossref (90) 26Caceres Payne M.R. Bender Rebhun Steward Differential tubulin immunocytochemistry hybridoma antibodies.J 1984; 4: 394-410Crossref confirmed co-staining PSD95 4D), noted vGlut1 often seemed so close almost overlap, whereas frequently adjacent distinct puncta; we, use mark enable straightforward identification tau. Using antibody (DA9) recognizes phosphorylated vGlut1-positive puncta (55.3%) MAP2-positive (70.2%) 63.3% 70.1% 4, C). With detect significant difference Under approximately 80% contain visible microtubules.27Gordon-Weeks Burgoyne Gray E.G. Presynaptic microtubules: organisation assembly/disassembly.Neuroscience. 1982; 739-749Abstract (46) Inasmuch 55.3% presynapses reasonable, possibly underestimate. criteria, neighboring structures exhibit 70.2% labeling, indicated localization.13Ittner Immunocytochemistry labeled substantial percentage synapses: 20.8% 32.0% 5). These support hypothesis already progression misfolding occur synapse. oligomers, solubilize free cytoplasmic proteins, remaining insoluble material crude PSD retained nearly all actin microtubule cytoskeleton 6). Most extractable Triton, small amount co-immunoprecipitated PSD95.13Ittner essentially Triton-insoluble indicating supports spines. ubiquitin-

Language: Английский

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Tau association with synaptic vesicles causes presynaptic dysfunction

Nature Communications, Journal Year: 2017, Volume and Issue: 8(1)

Published: May 11, 2017

Abstract Tau is implicated in more than 20 neurodegenerative diseases, including Alzheimer’s disease. Under pathological conditions, dissociates from axonal microtubules and missorts to pre- postsynaptic terminals. Patients suffer early synaptic dysfunction prior aggregate formation, but the underlying mechanism unclear. Here we show that pathogenic binds vesicles via its N-terminal domain interferes with presynaptic functions, vesicle mobility release rate, lowering neurotransmission fly rat neurons. Pathological mutants lacking binding still localize compartment do not impair function Moreover, an exogenously applied membrane-permeable peptide competes for Tau-vesicle suppresses Tau-induced toxicity Our work uncovers a role of may be part pathology various Tauopathies could exploited therapeutically.

Language: Английский

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Glial contributions to neurodegeneration in tauopathies

Molecular Neurodegeneration, Journal Year: 2017, Volume and Issue: 12(1)

Published: June 29, 2017

Tauopathies are a broad set of neurodegenerative dementias characterized by aggregation the tau protein into filamentous inclusions that can be found in neurons and glial cells. Activated microglia, astrocytes elevated levels proinflammatory molecules also pathological hallmarks brain regions affected pathology. There has been abundant research recent years to understand role gliosis neuroinflammation diseases, particularly Alzheimer's disease (AD) which is most common form dementia. AD tauopathy both extracellular amyloid-β plaques addition intracellular neurofibrillary tangles neuropil threads containing aggregated protein. Accumulating evidence suggests offers possible mechanistic link between these pathologies. Additionally, there appears for aggravating pathology neurodegeneration tauopathies featuring deposits as predominant signature. In this review, we survey literature regarding inflammatory mechanisms may impact related tauopathies. We consider physical microglia spread well non-cell autonomous effects secreted cytokines, specifically interleukin 1 beta, 6, tumor necrosis factor alpha complement proteins. These appear have direct on pathophysiology overall neuronal health. They indirectly homeostasis altering function. conclude proposing complex accelerating progression other

Language: Английский

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Apolipoprotein E Sets the Stage: Response to Injury Triggers Neuropathology

Neuron, Journal Year: 2012, Volume and Issue: 76(5), P. 871 - 885

Published: Dec. 1, 2012

Language: Английский

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The role of astrocytes in amyloid production and Alzheimer's disease

Open Biology, Journal Year: 2017, Volume and Issue: 7(12), P. 170228 - 170228

Published: Dec. 1, 2017

Alzheimer's disease (AD) is marked by the presence of extracellular amyloid beta (Aβ) plaques, intracellular neurofibrillary tangles (NFTs) and gliosis, activated glial cells, in brain. It thought that Aβ plaques trigger NFT formation, neuronal cell death, neuroinflammation gliosis and, ultimately, cognitive impairment. There are increased numbers reactive astrocytes AD, which surround secrete proinflammatory factors can phagocytize break down Aβ. was cells were major source However, mounting evidence suggests may play an additional role AD secreting significant quantities contributing to overall burden Astrocytes most numerous type brain, therefore even minor secretion from individual could prove be substantial when taken across whole Reactive have levels three necessary components for production: precursor protein, β-secretase (BACE1) γ-secretase. The identification environmental factors, such as neuroinflammation, promote astrocytic production, redefine how we think about developing therapeutics AD.

Language: Английский

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Frontotemporal Dementia

Neurologic Clinics, Journal Year: 2017, Volume and Issue: 35(2), P. 339 - 374

Published: April 11, 2017

Language: Английский

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mTOR regulates tau phosphorylation and degradation: implications for Alzheimer's disease and other tauopathies

Aging Cell, Journal Year: 2013, Volume and Issue: 12(3), P. 370 - 380

Published: Feb. 20, 2013

Accumulation of tau is a critical event in several neurodegenerative disorders, collectively known as tauopathies, which include Alzheimer's disease and frontotemporal dementia. Pathological hyperphosphorylated aggregates to form neurofibrillary tangles. The molecular mechanisms leading accumulation remain unclear more needs be done elucidate them. Age major risk factor for all suggesting that changes contributing the aging process may facilitate represent common across different tauopathies. Here, we use multiple animal models complementary genetic pharmacological approaches show mammalian target rapamycin (mTOR) regulates phosphorylation degradation. Specifically, genetically increasing mTOR activity elevates endogenous mouse levels phosphorylation. Complementary it, further demonstrate pharmacologically reducing signaling with ameliorates pathology associated behavioral deficits model overexpressing mutant human tau. Mechanistically, provide compelling evidence association between linked GSK3β autophagy function. In summary, facilitates pathology, while pathology. Given overwhelming increases lifespan healthspan, data presented here have profound clinical implications tauopathies basis how contribute Additionally, these results preclinical indicating valid therapeutic approach

Language: Английский

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