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Biochemical Isolation of Insoluble Tau in Transgenic Mouse Models of Tauopathies

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Amyloid Proteins

Part of the book series: Methods in Molecular Biology ((MIMB,volume 849))

Abstract

Tau is a highly soluble microtubule-associated protein (MAP) that is abundant in the central nervous system and expressed mainly in neuronal axons. Intracellular aggregates of insoluble tau protein are present in a group of neurodegenerative diseases called tauopathies, which include Alzheimer’s disease. Numerous transgenic mouse models of tauopathies have been produced in the last decade, and analysis of insoluble tau in these animals has provided a powerful tool to understand the development of tau pathology. In this short chapter, we aim at reviewing the two main isolation methods, sarkosyl and formic acid extraction (and their variations), used for the biochemical isolation of insoluble tau in transgenic mouse models of tauopathy, and discuss their advantages and drawbacks.

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References

  1. Weingarten, M. D., Lockwood, A. H., Hwo, S. Y., and Kirschner, M. W. (1975) A protein factor essential for microtubule assembly, Proc Natl Acad Sci USA 72, 1858–1862.

    Article  PubMed  CAS  Google Scholar 

  2. Cleveland, D. W., Hwo, S. Y., and Kirschner, M. W. (1977) Purification of tau, a microtubule-associated protein that induces assembly of microtubules from purified tubulin, J Mol Biol 116, 207–225.

    Article  PubMed  CAS  Google Scholar 

  3. Avila, J., Lucas, J. J., Perez, M., and Hernandez, F. (2004) Role of tau protein in both physiological and pathological conditions, Physiol Rev 84, 361–384.

    Article  PubMed  CAS  Google Scholar 

  4. Buee, L., Bussiere, T., Buee-Scherrer, V., Delacourte, A., and Hof, P. R. (2000) Tau protein isoforms, phosphorylation and role in neurodegenerative disorders, Brain Res. Brain. Res. Rev. 33, 95–130.

    Article  PubMed  CAS  Google Scholar 

  5. Alonso, A. C., Zaidi, T., Novak, M., Grundke-Iqbal, I., and Iqbal, K. (2001) Hyperphosphorylation induces self-assembly of tau into tangles of paired helical filaments / straight filaments, Proc. Natl. Acad. Sci. USA. 98, 6923–6928.

    Article  PubMed  CAS  Google Scholar 

  6. Sato, S., Tatebayashi, Y., Akagi, T., Chui, D. H., Murayama, M., Miyasaka, T., Planel, E., Tanemura, K., Sun, X., Hashikawa, T., Yoshioka, K., Ishiguro, K., and Takashima, A. (2002) Aberrant tau phosphorylation by glycogen synthase kinase-3beta and JNK3 induces oligomeric tau fibrils in COS-7 cells, J Biol Chem 277, 42060–42065.

    Article  PubMed  CAS  Google Scholar 

  7. Trojanowski, J. Q., and Lee, V. M. (1994) Paired helical filament tau in Alzheimer’s disease. The kinase connection, Am. J. Pathol. 144, 449–453.

    PubMed  CAS  Google Scholar 

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

    Article  PubMed  CAS  Google Scholar 

  9. Braak, H., and Braak, E. (1997) Frequency of stages of Alzheimer-related lesions in different age categories, Neurobiol. Aging 18, 351–357.

    CAS  Google Scholar 

  10. Delacourte, A., David, J. P., Sergeant, N., Buee, L., Wattez, A., Vermersch, P., Ghozali, F., Fallet-Bianco, C., Pasquier, F., Lebert, F., Petit, H., and Di Menza, C. (1999) The biochemical pathway of neurofibrillary degeneration in aging and Alzheimer’s disease, Neurology 52, 1158–1165.

    PubMed  CAS  Google Scholar 

  11. Arriagada, P. V., Growdon, J. H., Hedley-Whyte, E. T., and Hyman, B. T. (1992) Neurofibrillary tangles but not senile plaques parallel duration and severity of Alzheimer’s disease, Neurology 42, 631–639.

    PubMed  CAS  Google Scholar 

  12. Guillozet, A. L., Weintraub, S., Mash, D. C., and Mesulam, M. M. (2003) Neurofibrillary tangles, amyloid, and memory in aging and mild cognitive impairment, Arch Neurol 60, 729–736.

    Article  PubMed  Google Scholar 

  13. Wilcock, G. K., and Esiri, M. M. (1982) Plaques, tangles and dementia. A quantitative study, J Neurol Sci 56, 343–356.

    Article  PubMed  CAS  Google Scholar 

  14. Bretteville, A., and Planel, E. (2008) Tau aggregates: toxic, inert, or protective species?, J Alzheimers Dis 14, 431–436.

    PubMed  Google Scholar 

  15. Greenberg, S. G., and 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 

  16. Selkoe, D. J. (1986) Altered structural proteins in plaques and tangles: what do they tell us about the biology of Alzheimer’s disease?, Neurobiol Aging 7, 425–432.

    Article  PubMed  CAS  Google Scholar 

  17. Wisniewski, H. M., Iqbal, K., Grundke-Iqbal, I., and Rubenstein, R. (1987) The solubility controversy of paired helical filaments: a commentary, Neurochem Res 12, 93–95.

    Article  PubMed  CAS  Google Scholar 

  18. Ksiezak-Reding, H., Binder, L. I., and Yen, S. H. (1990) Alzheimer disease proteins (A68) share epitopes with tau but show distinct biochemical properties, J Neurosci Res 25, 420–430.

    Article  PubMed  CAS  Google Scholar 

  19. Lewis, J., McGowan, E., Rockwood, J., Melrose, H., Nacharaju, P., Van Slegtenhorst, M., Gwinn-Hardy, K., Paul Murphy, M., Baker, M., Yu, X., Duff, K., Hardy, J., Corral, A., Lin, W. L., Yen, S. H., Dickson, D. W., Davies, P., and Hutton, M. (2000) Neurofibrillary tangles, amyotrophy and progressive motor disturbance in mice expressing mutant (P301L) tau protein, Nat Genet 25, 402–405.

    Article  PubMed  CAS  Google Scholar 

  20. Sahara, N., Lewis, J., DeTure, M., McGowan, E., Dickson, D. W., Hutton, M., and Yen, S. H. (2002) Assembly of tau in transgenic animals expressing P301L tau: alteration of phosphorylation and solubility, J Neurochem 83, 1498–1508.

    Article  PubMed  CAS  Google Scholar 

  21. Eckermann, K., Mocanu, M. M., Khlistunova, I., Biernat, J., Nissen, A., Hofmann, A., Schonig, K., Bujard, H., Haemisch, A., Mandelkow, E., Zhou, L., Rune, G., and Mandelkow, E. M. (2007) The beta-propensity of Tau determines aggregation and synaptic loss in inducible mouse models of tauopathy, J Biol Chem 282, 31755–31765.

    Article  PubMed  CAS  Google Scholar 

  22. Noble, W., Planel, E., Zehr, C., Olm, V., Meyerson, J., Suleman, F., Gaynor, K., Wang, L., LaFrancois, J., Feinstein, B., Burns, M., Krishnamurthy, P., Wen, Y., Bhat, R., Lewis, J., Dickson, D., and Duff, K. (2005) Inhibition of glycogen synthase kinase-3 by lithium correlates with reduced tauopathy and degeneration in vivo, Proc Natl Acad Sci USA 102, 6990–6995.

    Article  PubMed  CAS  Google Scholar 

  23. Planel, E., Bretteville, A., Liu, L., Virag, L., Du, A. L., Yu, W. H., Dickson, D. W., Whittington, R. A., and Duff, K. E. (2009) Acceleration and persistence of neurofibrillary pathology in a mouse model of tauopathy following anesthesia, FASEB J 23, 2595–2604.

    Article  PubMed  CAS  Google Scholar 

  24. Skovronsky, D. M., Doms, R. W., and Lee, V. M. (1998) Detection of a novel intraneuronal pool of insoluble amyloid beta protein that accumulates with time in culture, J Cell Biol 141, 1031–1039.

    Article  PubMed  CAS  Google Scholar 

  25. Ishihara, T., Hong, M., Zhang, B., Nakagawa, Y., Lee, M. K., Trojanowski, J. Q., and Lee, V. M. (1999) Age-dependent emergence and progression of a tauopathy in transgenic mice overexpressing the shortest human tau isoform, Neuron 24, 751–762.

    Article  PubMed  CAS  Google Scholar 

  26. Kondo, J., Honda, T., Mori, H., Hamada, Y., Miura, R., Ogawara, M., and Ihara, Y. (1988) The carboxyl third of tau is tightly bound to paired helical filaments, Neuron 1, 827–834.

    Article  PubMed  CAS  Google Scholar 

  27. Lim, F., Hernandez, F., Lucas, J. J., Gomez-Ramos, P., Moran, M. A., and Avila, J. (2001) FTDP-17 mutations in tau transgenic mice provoke lysosomal abnormalities and Tau filaments in forebrain, Mol Cell Neurosci 18, 702–714.

    Article  PubMed  CAS  Google Scholar 

  28. Gotz, J., Chen, F., Barmettler, R., and Nitsch, R. M. (2001) Tau filament formation in transgenic mice expressing P301L tau, J Biol Chem 276, 529–534.

    Article  PubMed  CAS  Google Scholar 

  29. Tatebayashi, Y., Miyasaka, T., Chui, D. H., Akagi, T., Mishima, K., Iwasaki, K., Fujiwara, M., Tanemura, K., Murayama, M., Ishiguro, K., Planel, E., Sato, S., Hashikawa, T., and Takashima, A. (2002) Tau filament formation and associative memory deficit in aged mice expressing mutant (R406W) human tau, Proc Natl Acad Sci USA 99, 13896–13901.

    Article  PubMed  CAS  Google Scholar 

  30. Allen, B., Ingram, E., Takao, M., Smith, M. J., Jakes, R., Virdee, K., Yoshida, H., Holzer, M., Craxton, M., Emson, P. C., Atzori, C., Migheli, A., Crowther, R. A., Ghetti, B., Spillantini, M. G., and Goedert, M. (2002) Abundant tau filaments and nonapoptotic neurodegeneration in transgenic mice expressing human P301S tau protein, J Neurosci 22, 9340–9351.

    PubMed  CAS  Google Scholar 

  31. Andorfer, C., Kress, Y., Espinoza, M., de Silva, R., Tucker, K. L., Barde, Y. A., Duff, K., and Davies, P. (2003) Hyperphosphorylation and aggregation of tau in mice expressing normal human tau isoforms, J Neurochem 86, 582–590.

    Article  PubMed  CAS  Google Scholar 

  32. Ikeda, M., Shoji, M., Kawarai, T., Kawarabayashi, T., Matsubara, E., Murakami, T., Sasaki, A., Tomidokoro, Y., Ikarashi, Y., Kuribara, H., Ishiguro, K., Hasegawa, M., Yen, S. H., Chishti, M. A., Harigaya, Y., Abe, K., Okamoto, K., St George-Hyslop, P., and Westaway, D. (2005) Accumulation of filamentous tau in the cerebral cortex of human tau R406W transgenic mice, Am J Pathol 166, 521–531.

    Article  PubMed  CAS  Google Scholar 

  33. Santacruz, K., Lewis, J., Spires, T., Paulson, J., Kotilinek, L., Ingelsson, M., Guimaraes, A., DeTure, M., Ramsden, M., McGowan, E., Forster, C., Yue, M., Orne, J., Janus, C., Mariash, A., Kuskowski, M., Hyman, B., Hutton, M., and Ashe, K. H. (2005) Tau suppression in a neurodegenerative mouse model improves memory function, Science 309, 476–481.

    Article  PubMed  CAS  Google Scholar 

  34. Terwel, D., Lasrado, R., Snauwaert, J., Vandeweert, E., Van Haesendonck, C., Borghgraef, P., and Van Leuven, F. (2005) Changed conformation of mutant Tau-P301L underlies the moribund tauopathy, absent in progressive, nonlethal axonopathy of Tau-4R/2N transgenic mice, J Biol Chem 280, 3963–3973.

    Article  PubMed  CAS  Google Scholar 

  35. Murakami, T., Paitel, E., Kawarabayashi, T., Ikeda, M., Chishti, M. A., Janus, C., Matsubara, E., Sasaki, A., Kawarai, T., Phinney, A. L., Harigaya, Y., Horne, P., Egashira, N., Mishima, K., Hanna, A., Yang, J., Iwasaki, K., Takahashi, M., Fujiwara, M., Ishiguro, K., Bergeron, C., Carlson, G. A., Abe, K., Westaway, D., St George-Hyslop, P., and Shoji, M. (2006) Cortical neuronal and glial pathology in TgTauP301L transgenic mice: neuronal degeneration, memory disturbance, and phenotypic variation, Am J Pathol 169, 1365–1375.

    Article  PubMed  CAS  Google Scholar 

  36. Leroy, K., Bretteville, A., Schindowski, K., Gilissen, E., Authelet, M., De Decker, R., Yilmaz, Z., Buee, L., and Brion, J. P. (2007) Early axonopathy preceding neurofibrillary tangles in mutant tau transgenic mice, Am J Pathol 171, 976–992.

    Article  PubMed  CAS  Google Scholar 

  37. Rosenmann, H., Grigoriadis, N., Eldar-Levy, H., Avital, A., Rozenstein, L., Touloumi, O., Behar, L., Ben-Hur, T., Avraham, Y., Berry, E., Segal, M., Ginzburg, I., and Abramsky, O. (2008) A novel transgenic mouse expressing double mutant tau driven by its natural promoter exhibits tauopathy characteristics, Exp Neurol 212, 71–84.

    Article  PubMed  CAS  Google Scholar 

  38. Mocanu, M. M., Nissen, A., Eckermann, K., Khlistunova, I., Biernat, J., Drexler, D., Petrova, O., Schonig, K., Bujard, H., Mandelkow, E., Zhou, L., Rune, G., and Mandelkow, E. M. (2008) The potential for beta-structure in the repeat domain of tau protein determines aggregation, synaptic decay, neuronal loss, and coassembly with endogenous Tau in inducible mouse models of tauopathy, J Neurosci 28, 737–748.

    Article  PubMed  CAS  Google Scholar 

  39. Ittner, L. M., Fath, T., Ke, Y. D., Bi, M., van Eersel, J., Li, K. M., Gunning, P., and Gotz, J. (2008) Parkinsonism and impaired axonal transport in a mouse model of frontotemporal dementia, Proc Natl Acad Sci U S A 105, 15997–16002.

    Article  PubMed  CAS  Google Scholar 

  40. Adams, S. J., Crook, R. J., Deture, M., Randle, S. J., Innes, A. E., Yu, X. Z., Lin, W. L., Dugger, B. N., McBride, M., Hutton, M., Dickson, D. W., and McGowan, E. (2009) Overexpression of wild-type murine tau results in progressive tauopathy and neurodegeneration, Am J Pathol 175, 1598–1609.

    Article  PubMed  CAS  Google Scholar 

  41. Higuchi, M., Ishihara, T., Zhang, B., Hong, M., Andreadis, A., Trojanowski, J., and Lee, V. (2002) Transgenic mouse model of tauopathies with glial pathology and nervous system degeneration, Neuron 35, 433.

    Article  PubMed  CAS  Google Scholar 

  42. Zhang, B., Higuchi, M., Yoshiyama, Y., Ishihara, T., Forman, M. S., Martinez, D., Joyce, S., Trojanowski, J. Q., and Lee, V. M. (2004) Retarded axonal transport of R406W mutant tau in transgenic mice with a neurodegenerative tauopathy, J Neurosci 24, 4657–4667.

    Article  PubMed  CAS  Google Scholar 

  43. Forman, M. S., Lal, D., Zhang, B., Dabir, D. V., Swanson, E., Lee, V. M., and Trojanowski, J. Q. (2005) Transgenic mouse model of tau pathology in astrocytes leading to nervous system degeneration, J Neurosci 25, 3539–3550.

    Article  PubMed  CAS  Google Scholar 

  44. Higuchi, M., Zhang, B., Forman, M. S., Yoshiyama, Y., Trojanowski, J. Q., and Lee, V. M. (2005) Axonal degeneration induced by targeted expression of mutant human tau in oligodendrocytes of transgenic mice that model glial tauopathies, J Neurosci 25, 9434–9443.

    Article  PubMed  CAS  Google Scholar 

  45. Yoshiyama, Y., Higuchi, M., Zhang, B., Huang, S. M., Iwata, N., Saido, T. C., Maeda, J., Suhara, T., Trojanowski, J. Q., and Lee, V. M. (2007) Synapse loss and microglial activation precede tangles in a P301S tauopathy mouse model, Neuron 53, 337–351.

    Article  PubMed  CAS  Google Scholar 

  46. Probst, A., Gotz, J., Wiederhold, K. H., Tolnay, M., Mistl, C., Jaton, A. L., Hong, M., Ishihara, T., Lee, V. M., Trojanowski, J. Q., Jakes, R., Crowther, R. A., Spillantini, M. G., Burki, K., and Goedert, M. (2000) Axonopathy and amyotrophy in mice transgenic for human four-repeat tau protein, Acta Neuropathol 99, 469–481.

    Article  PubMed  CAS  Google Scholar 

  47. Gotz, J., Tolnay, M., Barmettler, R., Chen, F., Probst, A., and Nitsch, R. M. (2001) Oligodendroglial tau filament formation in transgenic mice expressing G272V tau, Eur J Neurosci 13, 2131–2140.

    Article  PubMed  CAS  Google Scholar 

  48. Tanemura, K., Murayama, M., Akagi, T., Hashikawa, T., Tominaga, T., Ichikawa, M., Yamaguchi, H., and Takashima, A. (2002) Neurodegeneration with tau accumulation in a transgenic mouse expressing V337M human tau, J. Neurosci. 22, 133–141.

    PubMed  CAS  Google Scholar 

  49. Perez, M., Valpuesta, J. M., de Garcini, E. M., Quintana, C., Arrasate, M., Lopez Carrascosa, J. L., Rabano, A., Garcia de Yebenes, J., and Avila, J. (1998) Ferritin is associated with the aberrant tau filaments present in progressive supranuclear palsy, Am J Pathol 152, 1531–1539.

    PubMed  CAS  Google Scholar 

  50. Goedert, M., Spillantini, M. G., Cairns, N. J., and Crowther, R. A. (1992) Tau proteins of Alzheimer paired helical filaments: abnormal phosphorylation of all six brain isoforms, Neuron 8, 159–168.

    Article  PubMed  CAS  Google Scholar 

  51. Oddo, S., Vasilevko, V., Caccamo, A., Kitazawa, M., Cribbs, D. H., and LaFerla, F. M. (2006) Reduction of soluble abeta and tau, but not soluble abeta alone, ameliorates cognitive decline in transgenic mice with plaques and tangles, J Biol Chem 281, 39413–39423.

    Article  PubMed  CAS  Google Scholar 

  52. Dabir, D. V., Robinson, M. B., Swanson, E., Zhang, B., Trojanowski, J. Q., Lee, V. M., and Forman, M. S. (2006) Impaired glutamate transport in a mouse model of tau pathology in astrocytes, J Neurosci 26, 644–654.

    Article  PubMed  CAS  Google Scholar 

  53. Brion, J. P., Hanger, D. P., Bruce, M. T., Couck, A. M., Flament-Durand, J., and Anderton, B. H. (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, 127–133.

    PubMed  CAS  Google Scholar 

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Acknowledgments

We are grateful to Dr. Peter Davies (Albert Einstein College of Medicine, Bronx, NY, USA) for helpful comments. This work was made possible by grants from the CIHR, FRSQ, NSERC, RQRV/ AFIRMAQ and FCI (to EP), and by a postdoctoral award from Alzheimer Society of Saskatchewan and from Alzheimer Society of Canada (to CJ). We apologize in advance if we missed an essential reference in this review.

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  1. Axe Neurosciences, Centre Hospitalier de l’Université Laval, Québec, QC, Canada

    Carl Julien, Alexis Bretteville & Emmanuel Planel

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  1. Carl Julien
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  2. Alexis Bretteville
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  3. Emmanuel Planel
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Correspondence to Emmanuel Planel .

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Editors and Affiliations

  1. School of Medicine, Dept. Psychiatry, New York University, First Ave. 550, New York, 10016, New York, USA

    Einar M. Sigurdsson

  2. Centro Nacional de Microbiología, Instituto de Salud Carlos III, Carretera Majadahonda-Pozuelo Km 2, Madrid, 28220, Spain

    Miguel Calero

  3. Inst. Química Física, Rocasolano, CSIC Madrid, C/ Serrano 119, Madrid, 28006, Spain

    María Gasset

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Julien, C., Bretteville, A., Planel, E. (2012). Biochemical Isolation of Insoluble Tau in Transgenic Mouse Models of Tauopathies. In: Sigurdsson, E., Calero, M., Gasset, M. (eds) Amyloid Proteins. Methods in Molecular Biology, vol 849. Humana Press. https://doi.org/10.1007/978-1-61779-551-0_32

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