Science China Life Sciences

, Volume 57, Issue 4, pp 422–431 | Cite as

Visualizing the microtubule-associated protein tau in the nucleus

  • Jing Lu
  • Ting Li
  • RongQiao He
  • Perry F. Bartlett
  • Jürgen GötzEmail author
Open Access
Research Paper Thematic Issue: From Brain Function to Therapy


Although tau is mainly known as an axonal microtubule-associated protein, many studies indicate that it is not restricted to this subcellular compartment. Assessing tau’s subcellular distribution, however, is not trivial as is evident from transgenic mouse studies. When human tau is over-expressed, it can be immunohistochemically localized to axons and the somatodendritic domain, modeling what is found in neurodegenerative diseases such as Alzheimer’s disease. Yet, in wild-type mice, despite its abundance, tau is difficult to visualize even in the axon. It is even more challenging to detect this protein in the nucleus, where tau has been proposed to protect DNA from damage. To establish a framework for future studies into tau’s nuclear functions, we compared several methods to visualize endogenous nuclear tau in cell lines and mouse brain. While depending on the fixation and permeabilization protocol, we were able to detect nuclear tau in SH-SY5Y human neuroblastoma cells, we failed to do so in N2a murine neuroblastoma cells. As a second method we used subcellular fractionation of mouse tissue and found that in the nucleus tau is mainly present in a hypophosphorylated form. When either full-length or truncated human tau was expressed, both accumulated in the cytoplasm, but were also found in the nuclear fraction. Because subcellular fractionation methods have their limitations, we finally isolated nuclei to probe for nuclear tau and found that the nuclei were free of cytoplasmic contamination. Together our analysis identifies several protocols for detecting tau in the nucleus where it is found in a less phosphorylated form.


Alzheimer’s disease fractionation microtubule-associated protein neuroblastoma nucleus phosphorylation tau 


  1. 1.
    Wang JZ, Liu F. Microtubule-associated protein tau in development, degeneration and protection of neurons. Prog Neurobiol, 2008, 85: 148–175PubMedCrossRefGoogle Scholar
  2. 2.
    Dehmelt L, Halpain S. The MAP2/Tau family of microtubule-associated proteins. Genome Biol, 2005, 6: 204PubMedPubMedCentralCrossRefGoogle Scholar
  3. 3.
    Chew YL, Fan X, Götz J, Nicholas HR. Protein with tau-like repeats regulates neuronal integrity and lifespan in C. elegans. J Cell Sci, 2013, 126: 2079–2091PubMedPubMedCentralCrossRefGoogle Scholar
  4. 4.
    Ittner LM, Ke YD, Delerue F, Bi M, Gladbach A, van Eersel J, Wolfing H, Chieng BC, Christie MJ, Napier IA, Eckert A, Staufenbiel M, Hardeman E, Götz J. Dendritic function of tau mediates amyloid-beta toxicity in Alzheimer’s disease mouse models, Cell, 2010, 142: 387–397PubMedCrossRefGoogle Scholar
  5. 5.
    Loomis PA, Howard TH, Castleberry RP, Binder LI. Identification of nuclear tau isoforms in human neuroblastoma cells. Proc Natl Acad Sci USA, 1990, 87: 8422–8426PubMedPubMedCentralCrossRefGoogle Scholar
  6. 6.
    Bai B, Hales CM, Chen PC, Gozal Y, Dammer EB, Fritz JJ, Wang X, Xia Q, Duong DM, Street C, Cantero G, Cheng D, Jones DR, Wu Z, Li Y, Diner I, Heilman CJ, Rees HD, Wu H, Lin L, Szulwach KE, Gearing M, Mufson EJ, Bennett DA, Montine TJ, Seyfried NT, Wingo TS, Sun YE, Jin P, Hanfelt J, Willcock DM, Levey A, Lah JJ, Peng J. U1 small nuclear ribonucleoprotein complex and RNA splicing alterations in Alzheimer’s disease. Proc Natl Acad Sci USA, 2013, 110: 16562–16567PubMedPubMedCentralCrossRefGoogle Scholar
  7. 7.
    Shea TB, Cressman CM. A 26–30 kDa developmentally-regulated tau isoform localized within nuclei of mitotic human neuroblastoma cells. Int J Dev Neurosci, 1998, 16: 41–48PubMedCrossRefGoogle Scholar
  8. 8.
    Wang Y, Loomis PA, Zinkowski RP, Binder LI. A novel tau transcript in cultured human neuroblastoma cells expressing nuclear tau. J Cell Biol, 1993, 121: 257–267PubMedCrossRefGoogle Scholar
  9. 9.
    Brady RM, Zinkowski RP, Binder LI. Presence of tau in isolated nuclei from human brain. Neurobiol Aging, 1995, 16: 479–486PubMedCrossRefGoogle Scholar
  10. 10.
    Cross DC, Munoz JP, Hernandez P, Maccioni RB. Nuclear and cytoplasmic tau proteins from human nonneuronal cells share common structural and functional features with brain tau. J Cell Biochem, 2000, 78: 305–317PubMedCrossRefGoogle Scholar
  11. 11.
    Chen F, David D, Ferrari A, Götz J. Posttranslational modifications of tau-role in human tauopathies and modeling in transgenic animals. Curr Drug Targets, 2004, 5: 503–515PubMedCrossRefGoogle Scholar
  12. 12.
    Maas T, Eidenmuller J, Brandt R. Interaction of tau with the neural membrane cortex is regulated by phosphorylation at sites that are modified in paired helical filaments. J Biol Chem, 2000, 275: 15733–15740PubMedCrossRefGoogle Scholar
  13. 13.
    Sultan A, Nesslany F, Violet M, Begard S, Loyens A, Talahari S, Mansuroglu Z, Marzin D, Sergeant N, Humez S, Colin M, Bonnefoy E, Buee L, Galas MC. Nuclear tau, a key player in neuronal DNA protection. J Biol Chem, 2011, 286: 4566–4575PubMedPubMedCentralCrossRefGoogle Scholar
  14. 14.
    Sjoberg MK, Shestakova E, Mansuroglu Z, Maccioni RB, Bonnefoy E. Tau protein binds to pericentromeric DNA: a putative role for nuclear tau in nucleolar organization. J Cell Sci, 2006, 119: 2025–2034PubMedCrossRefGoogle Scholar
  15. 15.
    Lu J, Miao J, Su T, Liu Y, He R. Formaldehyde induces hyperphosphorylation and polymerization of Tau protein both in vitro and in vivo. Biochim Biophys Acta, 2013, 1830: 4102–4116PubMedCrossRefGoogle Scholar
  16. 16.
    Liu C, Götz J. Profiling murine tau with 0N, 1N and 2N isoform-specific antibodies in brain and peripheral organs reveals distinct subcellular localization, with the 1N isoform being enriched in the nucleus. PLoS ONE, 2013, 8: e84849PubMedPubMedCentralCrossRefGoogle Scholar
  17. 17.
    Guillemin I, Becker M, Ociepka K, Friauf E, Nothwang HG. A subcellular prefractionation protocol for minute amounts of mammalian cell cultures and tissue. Proteomics, 2005, 5: 35–45PubMedCrossRefGoogle Scholar
  18. 18.
    Lim YA, Giese M, Shepherd C, Halliday G, Kobayashi M, Takamatsu K, Staufenbiel M, Eckert A, Götz J. Role of hippocalcin in mediating Abeta toxicity. Biochim Biophys Acta, 2012, 1822: 1247–1257PubMedCrossRefGoogle Scholar
  19. 19.
    Götz J, Nitsch RM. Compartmentalized tau hyperphosphorylation and increased levels of kinases in transgenic mice. Neuroreport, 2001, 12: 2007–2016PubMedCrossRefGoogle Scholar
  20. 20.
    Tucker KL, Meyer M, Barde YA. Neurotrophins are required for nerve growth during development. Nat Neurosci, 2001, 4: 29–37PubMedCrossRefGoogle Scholar
  21. 21.
    Götz J, Chen F, van Dorpe J, Nitsch RM. Formation of neurofibrillary tangles in P301L tau transgenic mice induced by Abeta 42 fibrils. Science, 2001, 293: 1491–1495PubMedCrossRefGoogle Scholar
  22. 22.
    Schild A, Schmidt K, Lim YA, Ke Y, Ittner LM, Hemmings BA, Götz J. Altered levels of PP2A regulatory B/PR55 isoforms indicate role in neuronal differentiation. Int J Dev Neurosci, 2006, 24: 437–443PubMedCrossRefGoogle Scholar
  23. 23.
    Zhou XW, Gustafsson JA, Tanila H, Bjorkdahl C, Liu R, Winblad B, Pei JJ. Tau hyperphosphorylation correlates with reduced methylation of protein phosphatase 2A. Neurobiol Dis, 2008, 31: 386–394PubMedCrossRefGoogle Scholar
  24. 24.
    Köhler C, Dinekov M, Götz J. Active glycogen synthase kinase-3 and tau pathology-related tyrosine phosphorylation in pR5 human tau transgenic mice. Neurobiol Aging, 2013, 34: 1369–1379PubMedCrossRefGoogle Scholar
  25. 25.
    Yu Y, Run X, Liang Z, Li Y, Liu F, Liu Y, Iqbal K, Grundke-Iqbal I, Gong CX. Developmental regulation of tau phosphorylation, tau kinases, and tau phosphatases. J Neurochem, 2009, 108: 1480–1494PubMedPubMedCentralCrossRefGoogle Scholar
  26. 26.
    Ittner LM, Götz J. Amyloid-beta and tau-a toxic pas de deux in Alzheimer’s disease. Nat Rev Neurosci, 2011, 12: 65–72PubMedCrossRefGoogle Scholar
  27. 27.
    Wei Y, Qu MH, Wang XS, Chen L, Wang DL, Liu Y, Hua Q, He RQ. Binding to the minor groove of the double-strand, tau protein prevents DNA from damage by peroxidation. PLoS ONE, 2008, 3: e2600PubMedPubMedCentralCrossRefGoogle Scholar
  28. 28.
    Bretteville A, Marcouiller F, Julien C, El Khoury NB, Petry FR, Poitras I, Mouginot D, Levesque G, Hebert SS, Planel E. Hypothermia-induced hyperphosphorylation: a new model to study tau kinase inhibitors. Sci Rep, 2012, 2: 480PubMedPubMedCentralCrossRefGoogle Scholar
  29. 29.
    Arrasate M, Perez M, Avila J. Tau dephosphorylation at tau-1 site correlates with its association to cell membrane. Neurochem Res, 2000, 25: 43–50PubMedCrossRefGoogle Scholar
  30. 30.
    Rhee HW, Zou P, Udeshi ND, Martell JD, Mootha VK, Carr SA, Ting AY. Proteomic mapping of mitochondria in living cells via spatially restricted enzymatic tagging. Science, 2013, 339: 1328–1331PubMedPubMedCentralCrossRefGoogle Scholar
  31. 31.
    Thurston VC, Zinkowski RP, Binder LI. Tau as a nucleolar protein in human nonneural cells in vitro and in vivo. Chromosoma, 1996, 105: 20–30PubMedCrossRefGoogle Scholar
  32. 32.
    Hua Q, He RQ. Tau could protect DNA double helix structure. Biochim Biophys Acta, 2003, 1645: 205–211PubMedCrossRefGoogle Scholar
  33. 33.
    Ke Y, Dramiga J, Schutz U, Kril JJ, Ittner LM, Schroder H, Götz J. Tau-mediated nuclear depletion and cytoplasmic accumulation of SFPQ in Alzheimer’s and Pick’s disease. PLoS ONE, 2012, 7: e35678PubMedPubMedCentralCrossRefGoogle Scholar
  34. 34.
    Gong CX, Grundke-Iqbal I, Iqbal K. Targeting tau protein in Alzheimer’s disease. Drugs Aging, 2010, 27: 351–365PubMedCrossRefGoogle Scholar
  35. 35.
    van Eersel J, Ke YD, Liu X, Delerue F, Kril JJ, Götz J, Ittner LM. Sodium selenate mitigates tau pathology, neurodegeneration, and functional deficits in Alzheimer’s disease models. Proc Natl Acad Sci USA, 2010, 107: 13888–13893PubMedPubMedCentralCrossRefGoogle Scholar

Copyright information

© The Author(s) 2014

Authors and Affiliations

  • Jing Lu
    • 1
  • Ting Li
    • 2
  • RongQiao He
    • 2
  • Perry F. Bartlett
    • 1
  • Jürgen Götz
    • 1
    Email author
  1. 1.Clem Jones Centre for Ageing Dementia Research, Queensland Brain InstituteThe University of QueenslandBrisbaneAustralia
  2. 2.Institute of BiophysicsChinese Academy of SciencesBeijingChina

Personalised recommendations