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Clathrin adaptor CALM/PICALM is associated with neurofibrillary tangles and is cleaved in Alzheimer’s brains

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Abstract

PICALM, a clathrin adaptor protein, plays important roles in clathrin-mediated endocytosis in all cell types. Recently, genome-wide association studies identified single nucleotide polymorphisms in PICALM gene as genetic risk factors for late-onset Alzheimer disease (LOAD). We analysed by western blotting with several anti-PICALM antibodies the pattern of expression of PICALM in human brain extracts. We found that PICALM was abnormally cleaved in AD samples and that the level of the uncleaved 65–75 kDa full-length PICALM species was significantly decreased in AD brains. Cleavage of human PICALM after activation of endogenous calpain or caspase was demonstrated in vitro. Immunohistochemistry revealed that PICALM was associated in situ with neurofibrillary tangles, co-localising with conformationally abnormal and hyperphosphorylated tau in LOAD, familial AD and Down syndrome cases. PHF-tau proteins co-immunoprecipitated with PICALM. PICALM was highly expressed in microglia in LOAD. These observations suggest that PICALM is associated with the development of AD tau pathology. PICALM cleavage could contribute to endocytic dysfunction in AD.

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References

  1. Ahle S, Ungewickell E (1986) Purification and properties of a new clathrin assembly protein. EMBO J 5:3143–3149

    PubMed  CAS  Google Scholar 

  2. Anderton BH, Callahan L, Coleman P et al (1998) Dendritic changes in Alzheimer’s disease and factors that may underlie these changes. Prog Neurobiol 55:595–609

    Article  PubMed  CAS  Google Scholar 

  3. Ando K, Leroy K, Heraud C et al (2011) Accelerated human mutant tau aggregation by knocking out murine tau in a transgenic mouse model. Am J Pathol 178:803–816

    Article  PubMed  CAS  Google Scholar 

  4. Baig S, Joseph SA, Tayler H et al (2010) Distribution and expression of PICALM in Alzheimer disease. J Neuropathol Exp Neurol 69:1071–1077

    Article  PubMed  CAS  Google Scholar 

  5. Ball M, Braak H, Coleman P et al (1997) Consensus recommendations for the postmortem diagnosis of Alzheimer’s disease. The National Institute on Aging, and Reagan Institute Working Group on Diagnostic Criteria for the Neuropathological Assessment of Alzheimer’s Disease. Neurobiol Aging 18:S1–S2

    Article  Google Scholar 

  6. Bertram L, McQueen MB, Mullin K, Blacker D, Tanzi RE (2007) Systematic meta-analyses of Alzheimer disease genetic association studies: the AlzGene database. Nat Genet 39:17–23

    Article  PubMed  CAS  Google Scholar 

  7. Boettner DR, Chi RJ, Lemmon SK (2012) Lessons from yeast for clathrin-mediated endocytosis. Nat Cell Biol 14:2–10

    Article  CAS  Google Scholar 

  8. Bondareff W, Wischik CM, Novak M, Roth M (1991) Sequestration of tau by granulovacuolar degeneration in Alzheimer’s disease. Am J Pathol 139:641–647

    PubMed  CAS  Google Scholar 

  9. Bossers K, Wirz KT, Meerhoff GF et al (2010) Concerted changes in transcripts in the prefrontal cortex precede neuropathology in Alzheimer’s disease. Brain 133:3699–3723

    Article  PubMed  Google Scholar 

  10. Braak E, Braak H, Mandelkow EM (1994) A sequence of cytoskeleton changes related to the formation of neurofibrillary tangles and neuropil threads. Acta Neuropathol 87:554–567

    Article  PubMed  CAS  Google Scholar 

  11. Braak H, Braak E (1991) Neuropathological stageing of Alzheimer-related changes. Acta Neuropathol 82:239–259

    Article  PubMed  CAS  Google Scholar 

  12. Brion JP, Couck AM, Passareiro E, Flament-Durand J (1985) Neurofibrillary tangles of Alzheimer’s disease: an immunohistochemical study. J Submicrosc Cytol 17:89–96

    PubMed  CAS  Google Scholar 

  13. Brion JP, Hanger DP, Bruce MT, Couck AM, Flament-Durand J, Anderton BH (1991) Tau in Alzheimer neurofibrillary tangles. N- and C-terminal regions are differentially associated with paired helical filaments and the location of a putative abnormal phosphorylation site. Biochem J 273(Pt 1):127–133

    PubMed  CAS  Google Scholar 

  14. Brion JP, Hanger DP, Couck AM, Anderton BH (1991) A68 proteins in Alzheimer’s disease are composed of several tau isoforms in a phosphorylated state which affects their electrophoretic mobilities. Biochem J 279(Pt 3):831–836

    PubMed  CAS  Google Scholar 

  15. Brion JP, Power D, Hue D, Couck AM, Anderton BH, Flament-Durand J (1989) Heterogeneity of ubiquitin immunoreactivity in neurofibrillary tangles of Alzheimer’s disease. Neurochem Int 14:121–128

    Article  PubMed  CAS  Google Scholar 

  16. Bushlin I, Petralia RS, Wu F et al (2008) Clathrin assembly protein AP180 and CALM differentially control axogenesis and dendrite outgrowth in embryonic hippocampal neurons. J Neurosci 28:10257–10271

    Article  PubMed  CAS  Google Scholar 

  17. Canu N, Dus L, Barbato C et al (1998) Tau cleavage and dephosphorylation in cerebellar granule neurons undergoing apoptosis. J Neurosci 18:7061–7074

    PubMed  CAS  Google Scholar 

  18. Cataldo AM, Peterhoff CM, Troncoso JC, Gomez-Isla T, Hyman BT, Nixon RA (2000) Endocytic pathway abnormalities precede amyloid beta deposition in sporadic Alzheimer’s disease and Down syndrome: differential effects of APOE genotype and presenilin mutations. Am J Pathol 157:277–286

    Article  PubMed  CAS  Google Scholar 

  19. Coleman PD, Yao PJ (2003) Synaptic slaughter in Alzheimer’s disease. Neurobiol Aging 24:1023–1027

    Article  PubMed  CAS  Google Scholar 

  20. Cotman CW, Poon WW, Rissman RA, Blurton-Jones M (2005) The role of caspase cleavage of tau in Alzheimer disease neuropathology. J Neuropathol Exp Neurol 64:104–112

    PubMed  CAS  Google Scholar 

  21. Dickson DW, Ksiezak-Reding H, Davies P, Yen SH (1987) A monoclonal antibody that recognizes a phosphorylated epitope in Alzheimer neurofibrillary tangles, neurofilaments and tau proteins immunostains granulovacuolar degeneration. Acta Neuropathol 73:254–258

    Article  PubMed  CAS  Google Scholar 

  22. Dourlen P, Ando K, Hamdane M, Begard S, Buee L, Galas MC (2007) The peptidyl prolyl cis/trans isomerase Pin1 downregulates the Inhibitor of Apoptosis Protein Survivin. Biochim Biophys Acta 1773:1428–1437

    Article  PubMed  CAS  Google Scholar 

  23. Dreyling MH, Martinez-Climent JA, Zheng M, Mao J, Rowley JD, Bohlander SK (1996) The t(10;11)(p13;q14) in the U937 cell line results in the fusion of the AF10 gene and CALM, encoding a new member of the AP-3 clathrin assembly protein family. Proc Natl Acad Sci USA 93:4804–4809

    Article  PubMed  CAS  Google Scholar 

  24. Ford MG, Pearse BM, Higgins MK et al (2001) Simultaneous binding of PtdIns(4,5)P2 and clathrin by AP180 in the nucleation of clathrin lattices on membranes. Science 291:1051–1055

    Article  PubMed  CAS  Google Scholar 

  25. Gamblin TC, Chen F, Zambrano A et al (2003) Caspase cleavage of tau: linking amyloid and neurofibrillary tangles in Alzheimer’s disease. Proc Natl Acad Sci USA 100:10032–10037

    Article  PubMed  CAS  Google Scholar 

  26. George JM, Jin H, Woods WS, Clayton DF (1995) Characterization of a novel protein regulated during the critical period for song learning in the zebra finch. Neuron 15:361–372

    Article  PubMed  CAS  Google Scholar 

  27. Gervais FG, Xu D, Robertson GS et al (1999) Involvement of caspases in proteolytic cleavage of Alzheimer’s amyloid-beta precursor protein and amyloidogenic A beta peptide formation. Cell 97:395–406

    Article  PubMed  CAS  Google Scholar 

  28. Greenberg SG, Davies P (1990) A preparation of Alzheimer paired helical filaments that displays distinct tau proteins by polyacrylamide gel electrophoresis. Proc Natl Acad Sci USA 87:5827–5831

    Article  PubMed  CAS  Google Scholar 

  29. Harold D, Abraham R, Hollingworth P et al (2009) Genome-wide association study identifies variants at CLU and PICALM associated with Alzheimer’s disease. Nat Genet 41:1088–1093

    Article  PubMed  CAS  Google Scholar 

  30. Hollingworth P, Harold D, Sims R et al (2011) Common variants at ABCA7, MS4A6A/MS4A4E, EPHA1, CD33 and CD2AP are associated with Alzheimer’s disease. Nat Genet 43:429–435

    Article  PubMed  CAS  Google Scholar 

  31. Hoozemans JJ, van Haastert ES, Nijholt DA, Rozemuller AJ, Eikelenboom P, Scheper W (2009) The unfolded protein response is activated in pretangle neurons in Alzheimer’s disease hippocampus. Am J Pathol 174:1241–1251

    Article  PubMed  CAS  Google Scholar 

  32. Joachim CL, Morris JH, Selkoe DJ, Kosik KS (1987) Tau epitopes are incorporated into a range of lesions in Alzheimer’s disease. J Neuropathol Exp Neurol 46:611–622

    Article  PubMed  CAS  Google Scholar 

  33. Jones L, Harold D, Williams J (2010) Genetic evidence for the involvement of lipid metabolism in Alzheimer’s disease. Biochim Biophys Acta 1801:754–761

    Article  PubMed  CAS  Google Scholar 

  34. Kamboh MI, Minster RL, Demirci FY et al (2012) Association of CLU and PICALM variants with Alzheimer’s disease. Neurobiol Aging 33:518–521

    Article  PubMed  CAS  Google Scholar 

  35. Karran E, Mercken M, De Strooper B (2011) The amyloid cascade hypothesis for Alzheimer’s disease: an appraisal for the development of therapeutics. Nat Rev Drug Discov 10:698–712

    Article  PubMed  CAS  Google Scholar 

  36. Kelly BL, Ferreira A (2006) beta-Amyloid-induced dynamin 1 degradation is mediated by N-methyl-D-aspartate receptors in hippocampal neurons. J Biol Chem 281:28079–28089

    Article  PubMed  CAS  Google Scholar 

  37. Kim JA, Kim HL (2001) Cleavage of purified neuronal clathrin assembly protein (CALM) by caspase 3 and calpain. Exp Mol Med 33:245–250

    Article  PubMed  CAS  Google Scholar 

  38. Koo SJ, Markovic S, Puchkov D et al (2012) SNARE motif-mediated sorting of synaptobrevin by the endocytic adaptors clathrin assembly lymphoid myeloid leukemia (CALM) and AP180 at synapses. Proc Natl Acad Sci USA 108:13540–13545

    Article  Google Scholar 

  39. Kunze G, Barre P, Scheidt HA, Thomas L, Eliezer D, Huster D (2012) Binding of the three-repeat domain of tau to phospholipid membranes induces an aggregated-like state of the protein. Biochim Biophys Acta 1818:2302–2313

    Article  PubMed  Google Scholar 

  40. Lacor PN, Buniel MC, Furlow PW et al (2007) Abeta oligomer-induced aberrations in synapse composition, shape, and density provide a molecular basis for loss of connectivity in Alzheimer’s disease. J Neurosci 27:796–807

    Article  PubMed  CAS  Google Scholar 

  41. Lambert JC, Heath S, Even G et al (2009) Genome-wide association study identifies variants at CLU and CR1 associated with Alzheimer’s disease. Nat Genet 41:1094–1099

    Article  PubMed  CAS  Google Scholar 

  42. Lambert JC, Zelenika D, Hiltunen M et al (2011) Evidence of the association of BIN1 and PICALM with the AD risk in contrasting European populations. Neurobiol Aging 32(756):e711–e755

    Google Scholar 

  43. Lee MS, Kao SC, Lemere CA et al (2003) APP processing is regulated by cytoplasmic phosphorylation. J Cell Biol 163:83–95

    Article  PubMed  CAS  Google Scholar 

  44. Lee MS, Kwon YT, Li M, Peng J, Friedlander RM, Tsai LH (2000) Neurotoxicity induces cleavage of p35 to p25 by calpain. Nature 405:360–364

    Article  PubMed  CAS  Google Scholar 

  45. Leroy K, Boutajangout A, Authelet M, Woodgett JR, Anderton BH, Brion JP (2002) The active form of glycogen synthase kinase-3beta is associated with granulovacuolar degeneration in neurons in Alzheimer’s disease. Acta Neuropathol 103:91–99

    Article  PubMed  CAS  Google Scholar 

  46. Luna-Munoz J, Chavez-Macias L, Garcia-Sierra F, Mena R (2007) Earliest stages of tau conformational changes are related to the appearance of a sequence of specific phospho-dependent tau epitopes in Alzheimer’s disease. J Alzheimers Dis 12:365–375

    PubMed  CAS  Google Scholar 

  47. Mena R, Edwards P, Perez-Olvera O, Wischik CM (1995) Monitoring pathological assembly of tau and beta-amyloid proteins in Alzheimer’s disease. Acta Neuropathol 89:50–56

    Article  PubMed  CAS  Google Scholar 

  48. Meyerholz A, Hinrichsen L, Groos S, Esk PC, Brandes G, Ungewickell EJ (2005) Effect of clathrin assembly lymphoid myeloid leukemia protein depletion on clathrin coat formation. Traffic 6:1225–1234

    Article  PubMed  CAS  Google Scholar 

  49. Miller SE, Sahlender DA, Graham SC et al (2011) The molecular basis for the endocytosis of small R-SNAREs by the clathrin adaptor CALM. Cell 147:1118–1131

    Article  PubMed  CAS  Google Scholar 

  50. Morgan JR, Zhao X, Womack M, Prasad K, Augustine GJ, Lafer EM (1999) A role for the clathrin assembly domain of AP180 in synaptic vesicle endocytosis. J Neurosci 19:10201–10212

    PubMed  CAS  Google Scholar 

  51. Naj AC, Jun G, Beecham GW et al (2011) Common variants at MS4A4/MS4A6E, CD2AP, CD33 and EPHA1 are associated with late-onset Alzheimer’s disease. Nat Genet 43:436–441

    Article  PubMed  CAS  Google Scholar 

  52. Nakagawa T, Zhu H, Morishima N et al (2000) Caspase-12 mediates endoplasmic-reticulum-specific apoptosis and cytotoxicity by amyloid-beta. Nature 403:98–103

    Article  PubMed  CAS  Google Scholar 

  53. Nicholson AM, Ferreira A (2009) Increased membrane cholesterol might render mature hippocampal neurons more susceptible to beta-amyloid-induced calpain activation and tau toxicity. J Neurosci 29:4640–4651

    Article  PubMed  CAS  Google Scholar 

  54. Nixon RA (2005) Endosome function and dysfunction in Alzheimer’s disease and other neurodegenerative diseases. Neurobiol Aging 26:373–382

    Article  PubMed  CAS  Google Scholar 

  55. Petralia RS, Wang YX, Indig FE et al (2012) Reduction of AP180 and CALM produces defects in synaptic vesicle size and density. Neuromolecular Med 15:49–60

    Google Scholar 

  56. Petralia RS, Yao PJ (2007) AP180 and CALM in the developing hippocampus: expression at the nascent synapse and localization to trafficking organelles. J Comp Neurol 504:314–327

    Article  PubMed  CAS  Google Scholar 

  57. Ronca F, Chan SL, Yu VC (1997) 1-(5-Isoquinolinesulfonyl)-2-methylpiperazine induces apoptosis in human neuroblastoma cells, SH-SY5Y, through a p53-dependent pathway. J Biol Chem 272:4252–4260

    Article  PubMed  CAS  Google Scholar 

  58. Rudinskiy N, Grishchuk Y, Vaslin A et al (2009) Calpain hydrolysis of alpha- and beta2-adaptins decreases clathrin-dependent endocytosis and may promote neurodegeneration. J Biol Chem 284:12447–12458

    Article  PubMed  CAS  Google Scholar 

  59. Saito K, Elce JS, Hamos JE, Nixon RA (1993) Widespread activation of calcium-activated neutral proteinase (calpain) in the brain in Alzheimer disease: a potential molecular basis for neuronal degeneration. Proc Natl Acad Sci USA 90:2628–2632

    Article  PubMed  CAS  Google Scholar 

  60. Schenk D, Barbour R, Dunn W et al (1999) Immunization with amyloid-beta attenuates Alzheimer-disease-like pathology in the PDAPP mouse. Nature 400:173–177

    Article  PubMed  CAS  Google Scholar 

  61. Seshadri S, Fitzpatrick AL, Ikram MA et al (2010) Genome-wide analysis of genetic loci associated with Alzheimer disease. JAMA 303:1832–1840

    Article  PubMed  CAS  Google Scholar 

  62. Sousa R, Tannery NH, Zhou S, Lafer EM (1992) Characterization of a novel synapse-specific protein. I. Developmental expression and cellular localization of the F1–20 protein and mRNA. J Neurosci 12:2130–2143

    PubMed  CAS  Google Scholar 

  63. Suzuki M, Tanaka H, Tanimura A et al (2012) The clathrin assembly protein PICALM is required for erythroid maturation and transferrin internalization in mice. PLoS ONE 7:e31854

    Article  PubMed  CAS  Google Scholar 

  64. Sze CI, Bi H, Kleinschmidt-DeMasters BK, Filley CM, Martin LJ (2000) Selective regional loss of exocytotic presynaptic vesicle proteins in Alzheimer’s disease brains. J Neurol Sci 175:81–90

    Article  PubMed  CAS  Google Scholar 

  65. Tebar F, Bohlander SK, Sorkin A (1999) Clathrin assembly lymphoid myeloid leukemia (CALM) protein: localization in endocytic-coated pits, interactions with clathrin, and the impact of overexpression on clathrin-mediated traffic. Mol Biol Cell 10:2687–2702

    PubMed  CAS  Google Scholar 

  66. Thal DR, Rub U, Orantes M, Braak H (2002) Phases of A beta-deposition in the human brain and its relevance for the development of AD. Neurology 58:1791–1800

    Article  PubMed  Google Scholar 

  67. Thomas RS, Lelos MJ, Good MA, Kidd EJ (2011) Clathrin-mediated endocytic proteins are upregulated in the cortex of the Tg2576 mouse model of Alzheimer’s disease-like amyloid pathology. Biochem Biophys Res Commun 415:656–661

    PubMed  CAS  Google Scholar 

  68. Treusch S, Hamamichi S, Goodman JL et al (2011) Functional links between Abeta toxicity, endocytic trafficking, and Alzheimer’s disease risk factors in yeast. Science 334:1241–1245

    Article  PubMed  CAS  Google Scholar 

  69. Troy CM, Rabacchi SA, Friedman WJ, Frappier TF, Brown K, Shelanski ML (2000) Caspase-2 mediates neuronal cell death induced by beta-amyloid. J Neurosci 20:1386–1392

    PubMed  CAS  Google Scholar 

  70. Uchihara T, Nakamura A, Yamazaki M, Mori O (2000) Tau-positive neurons in corticobasal degeneration and Alzheimer’s disease–distinction by thiazin red and silver impregnations. Acta Neuropathol 100:385–389

    Article  PubMed  CAS  Google Scholar 

  71. Vingtdeux V, Davies P, Dickson DW, Marambaud P (2011) AMPK is abnormally activated in tangle- and pre-tangle-bearing neurons in Alzheimer’s disease and other tauopathies. Acta Neuropathol 121:337–349

    Article  PubMed  CAS  Google Scholar 

  72. Wu F, Mattson MP, Yao PJ (2010) Neuronal activity and the expression of clathrin-assembly protein AP180. Biochem Biophys Res Commun 402:297–300

    Article  PubMed  CAS  Google Scholar 

  73. Wu Y, Liang S, Oda Y et al (2007) Truncations of amphiphysin I by calpain inhibit vesicle endocytosis during neural hyperexcitation. EMBO J 26:2981–2990

    Article  PubMed  CAS  Google Scholar 

  74. Xiao Q, Gil SC, Yan P et al (2012) Role of phosphatidylinositol clathrin assembly lymphoid-myeloid leukemia (PICALM) in intracellular amyloid precursor protein (APP) processing and amyloid plaque pathogenesis. J Biol Chem 287:21279–21289

    Article  PubMed  CAS  Google Scholar 

  75. Yamazaki Y, Matsubara T, Takahashi T et al (2011) Granulovacuolar degenerations appear in relation to hippocampal phosphorylated tau accumulation in various neurodegenerative disorders. PLoS ONE 6:e26996

    Article  PubMed  CAS  Google Scholar 

  76. Yao PJ, Coleman PD, Calkins DJ (2002) High-resolution localization of clathrin assembly protein AP180 in the presynaptic terminals of mammalian neurons. J Comp Neurol 447:152–162

    Article  PubMed  CAS  Google Scholar 

  77. Yao PJ, Petralia RS, Bushlin I, Wang Y, Furukawa K (2005) Synaptic distribution of the endocytic accessory proteins AP180 and CALM. J Comp Neurol 481:58–69

    Article  PubMed  CAS  Google Scholar 

  78. Yao PJ, Zhu M, Pyun EI et al (2003) Defects in expression of genes related to synaptic vesicle trafficking in frontal cortex of Alzheimer’s disease. Neurobiol Dis 12:97–109

    Article  PubMed  CAS  Google Scholar 

  79. Ye W, Lafer EM (1995) Clathrin binding and assembly activities of expressed domains of the synapse-specific clathrin assembly protein AP-3. J Biol Chem 270:10933–10939

    Article  PubMed  CAS  Google Scholar 

  80. Zhang B, Koh YH, Beckstead RB, Budnik V, Ganetzky B, Bellen HJ (1998) Synaptic vesicle size and number are regulated by a clathrin adaptor protein required for endocytosis. Neuron 21:1465–1475

    Article  PubMed  CAS  Google Scholar 

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Acknowledgments

This study was supported by ANR Franco-Canada CholAD, INSERM (U975-P-A1L4-RP-RPV10014DDA-R10178DD), by MassImage, INSERM (U975-P-A1L4-RP-RPV10015DDA-R10179DD), by grants from dotation d’équipe, INSERM, and by “Investissements d’avenir” ANR-10-IAIHU-06 to C.D., as well as by grants from the Diane programme (Walloon region) (816856), FRMA/SAO, and the Fonds de la Recherche Scientifique Médicale (3.4504.10) to J.P.B. The brain tissues were kindly provided by the GIE NeuroCEB Brain Bank (France Alzheimer, France Parkinson, ARSEP, CSC). We thank Dr Peter Davies for providing anti-tau antibodies, Drs George and Clayton for anti-α synuclein antibody and Dr. Sabrina Turbant for human tissue preparation. Analysis by confocal microscopy was performed at the cellular imaging platform of Pitié-Salpêtrière PICPS.

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  1. Laboratoire de Neuropathologie Escourolle, Hôpital de la Pitié-Salpêtrière, AP-HP, Paris, France

    Kunie Ando, Anaïs Chanut, Véronique Sazdovitch & Charles Duyckaerts

  2. Group of Alzheimer’s and Prion’s diseases, ICM Research Centre, UPMC, INSERM UMR S 975, CNRS UMR 7225, Hôpital de la Pitié-Salpêtrière, 47 Bd de l’Hôpital, 75013, Paris, France

    Kunie Ando, Jérémie Lavaur, Marie-Claude Potier & Charles Duyckaerts

  3. Laboratory of Histology, Neuroanatomy and Neuropathology, UNI (ULB Neuroscience Institute), Université Libre de Bruxelles, 1070, Brussels, Belgium

    Jean-Pierre Brion, Virginie Stygelbout, Valérie Suain, Michèle Authelet & Robert Dedecker

  4. Neurobiology Department, Northwestern University, Evanston, IL, 60208, USA

    Pascale Lacor

  5. Centre for Research in Neurodegenerative Diseases, Faculty of Medicine, University of Toronto, Toronto, Canada

    Ekaterina Rogaeva

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401_2013_1111_MOESM1_ESM.jpg

Fig. S1: PICALM in control and AD brains detected with HPA019061 and sc-6433 anti-PICALM antibodies. a-b: Western blotting for PICALM in the RIPA-soluble and insoluble fractions detected with HPA019061 (a) and with sc-6433 (b). The 75 kDa PICALM isoform was decreased in RIPA-soluble fraction of AD brain lysate. 50 kDa PICALM fragment was detected with HPA019061 whereas 25 kDa fragment was detected with sc-6433. There was an unknown band at 95 kDa detected with sc-6433 in the RIPA-insoluble fraction (asterisk) (b). c: Western blot analysis of ADDLs for their potential cross-reactivity with PICALM antibodies. ADDLs were analysed by western blotting using the anti-Aβ (6E10) antibody and three different anti-PICALM antibodies as indicated. ADDLs were strongly immunoreactive with the 6E10 antibody but were not reactive with the anti-PICALM antibodies. (JPEG 579 kb)

401_2013_1111_MOESM2_ESM.jpg

Fig. S2: Immunostaining of PICALM with different anti-PICALM antibodies in control and Down syndrome brains. a-o: PICALM immunostaining with HPA019053 (a, d, g, j, m), HPA019061 (b, e, h, k, n) and sc-6433 (c, f, i, l, o) in endothelial cells (a-c), choroid plexus (d-f), neurons (g-r) in the hippocampus of control brains (a-i) and in the hippocampus of Down syndrome (j-r). PICALM labelling pattern in the endothelial cells and in the choroid plexus was quite similar with all the three anti-PICALM antibodies (a-f). There was non-specific staining of lipofuscin with sc-6433 (i, o). p-r: Anti-PICALM antibodies HPA019053 (p), HPA019061 (q) and sc-6433 (r) were immunoabsorbed with GST-PICALM in prior to immunostaining. Scale bars 20 μm. (JPEG 4283 kb)

401_2013_1111_MOESM3_ESM.jpg

Fig. S3: Absence of PICALM immunoreactivity on isolated PHFs present in sarkosyl insoluble fraction. a: Immunogold staining of isolated PHFs present in the sarkosyl insoluble fraction using anti-PICALM HPA019053 antibody. Isolated PHFs were not immunoreactive with the PICALM antibody. b: Positive control with the anti-tau PHF1 antibody (10 nm gold particles). c: Negative control without primary antibody. Scale bar 0.2 um. d: Western blot analysis of sarkosyl-soluble and insoluble fractions for tau (AT8) and PICALM (HPA019053). PHF-tau was present only in sarkosyl insoluble fraction of AD brains and was absent in control brains. PICALM was not co-enriched with PHF-tau by sarkosyl fraction. (JPEG 840 kb)

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Ando, K., Brion, JP., Stygelbout, V. et al. Clathrin adaptor CALM/PICALM is associated with neurofibrillary tangles and is cleaved in Alzheimer’s brains. Acta Neuropathol 125, 861–878 (2013). https://doi.org/10.1007/s00401-013-1111-z

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