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Cellular and Molecular Neurobiology

, Volume 36, Issue 4, pp 593–602 | Cite as

The Co-chaperone BAG2 Mediates Cold-Induced Accumulation of Phosphorylated Tau in SH-SY5Y Cells

  • Cesar Augusto Dias de Paula
  • Fernando Enrique Santiago
  • Adriele Silva Alves de Oliveira
  • Fernando Augusto Oliveira
  • Maria Camila Almeida
  • Daniel Carneiro CarrettieroEmail author
Original Research

Abstract

Inclusions of phosphorylated tau (p-tau) are a hallmark of many neurodegenerative disorders classified as “tauopathy,” of which Alzheimer’s disease is the most prevalent form. Dysregulation of tau phosphorylation disrupts neuron structure and function, and hyperphosphorylated tau aggregates to form neurotoxic inclusions. The abundance of ubiquitin in tau inclusions suggests a defect in ubiquitin-mediated tau protein degradation by the proteasome. Under the temperature of 37 °C, the co-chaperone BAG2 protein targets phosphorylated tau for degradation via by a more-efficient, ubiquitin-independent pathway. In both in vivo and in vitro studies, cold exposure induces the accumulation of phosphorylated tau protein. The SH-SY5Y cell line differentiates into neuron-like cells on treatment with retinoic acid and is an established model for research on the effects of cold on tau phosphorylation. The aim of the present study was to investigate whether BAG2 mediates the cold-induced accumulation of phosphorylated tau protein. Our findings show that cold exposure causes a decrease in BAG2 expression in undifferentiated cells. Conversely, BAG2 expression is increased in differentiated cells exposed to cold. Further, undifferentiated cells exposed to cold had an increased proportion of p-tau to total tau, suggesting an accumulation of p-tau that is consistent with decreased levels of BAG2. Overexpression of BAG2 in cold-exposed undifferentiated cells restored levels of p-tau to those of 37 °C undifferentiated control. Interestingly, although BAG2 expression increased in differentiated cells, this increase was not accompanied by a decrease in the proportion of p-tau to total tau. Further, overexpression of BAG2 in cold exposed differentiated cells showed no significant difference in p-tau levels compared to 37 °C controls. Taken together, these data show that expression of BAG2 is differently regulated in a differentiation-dependent context. Our results suggest that repression of BAG2 expression or BAG2 activity by cold-sensitive pathways, as modeled in undifferentiated and differentiated cells, respectively, may be a causal factor in the accumulation of cytotoxic hyperphosphorylated tau protein via restriction of BAG2-mediated clearance of cellular p-tau.

Keywords

Temperature Cold exposure Hypothermia Differentiation Cell Tauopathy Alzheimer’s disease 

Notes

Acknowledgments

The authors with to acknowledge extramural financial support provided by FAPESP (Grant, 2009/11446-4, 2011/06528-1 and 2012/50336-2) and CNPq (449102/2014-9) as well as financial support from CAPES and UFABC intramural funds.

Authors’ Contributions

C. A. D. P., M. C. A. and D. C. C. contributed to design of experiments. C. A. D. P., F. E. S., A. S. A. O., and F. A. O. performed experiments. Analysis of experimental data was done by C. A. D. P., F. E. S., M. C. A., and D. C. C. The manuscript was prepared by C. A. D. P., F. E. S. and D. C. C.

Compliance with Ethical Standards

Conflict of Interest

The authors declare no competing interests.

References

  1. Abramoff M, Magelhaes P, Ram S (2004) Image processing with ImageJ. Biophotonics Int 11:36–42Google Scholar
  2. Bendiske J, Caba E, Brown QB, Bahr BA (2002) Intracellular deposition, microtubule destabilization, and transport failure: an “early” pathogenic cascade leading to synaptic decline. J Neuropathol Exp Neurol 61:640–650CrossRefPubMedGoogle Scholar
  3. Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254CrossRefPubMedGoogle Scholar
  4. Bretteville A, Marcouiller F, Julien C, El Khoury NB, Petry FR, Poitras I, Mouginot D, Levesque G, Hebert SS, Planel E (2012) Hypothermia-induced hyperphosphorylation: a new model to study tau kinase inhibitors. Sci Rep 2:480CrossRefPubMedPubMedCentralGoogle Scholar
  5. Buee L, Bussiere T, Buee-Scherrer V, Delacourte A, Hof PR (2000) Tau protein isoforms, phosphorylation and role in neurodegenerative disorders. Brain Res Brain Res Rev 33:95–130CrossRefPubMedGoogle Scholar
  6. Carrettiero DC, Hernandez I, Neveu P, Papagiannakopoulos T, Kosik KS (2009) The cochaperone BAG2 sweeps paired helical filament-insoluble tau from the microtubule. J Neurosci 29:2151–2161CrossRefPubMedPubMedCentralGoogle Scholar
  7. Cheung YT, Lau WK, Yu MS, Lai CS, Yeung SC, So KF, Chang RC (2009) Effects of all-trans-retinoic acid on human SH-SY5Y neuroblastoma as in vitro model in neurotoxicity research. Neurotoxicology 30:127–135CrossRefPubMedGoogle Scholar
  8. Collins KJ, Exton-Smith AN, Dore C (1981) Urban hypothermia: preferred temperature and thermal perception in old age. Br Med J (Clin Res Ed) 282:175–177CrossRefGoogle Scholar
  9. Dalen ML, Froyland E, Saugstad OD, Mollnes TE, Rootwelt T (2009) Post-hypoxic hypothermia is protective in human NT2-N neurons regardless of oxygen concentration during reoxygenation. Brain Res 1259:80–89CrossRefPubMedGoogle Scholar
  10. Dong Y, Wu X, Xu Z, Zhang Y, Xie Z (2012) Anesthetic isoflurane increases phosphorylated tau levels mediated by caspase activation and Abeta generation. PLoS One 7:e39386CrossRefPubMedPubMedCentralGoogle Scholar
  11. Drewes G, Lichtenberg-Kraag B, Doring F, Mandelkow EM, Biernat J, Goris J, Doree M, Mandelkow E (1992) Mitogen activated protein (MAP) kinase transforms tau protein into an Alzheimer-like state. EMBO J 11:2131–2138PubMedPubMedCentralGoogle Scholar
  12. Feng Q, Cheng B, Yang R, Sun FY, Zhu CQ (2005) Dynamic changes of phosphorylated tau in mouse hippocampus after cold water stress. Neurosci Lett 388:13–16CrossRefPubMedGoogle Scholar
  13. Inoue Y, Nakao M, Araki T, Ueda H (1992) Thermoregulatory responses of young and older men to cold exposure. Eur J Appl Physiol Occup Physiol 65:492–498CrossRefPubMedGoogle Scholar
  14. Iqbal K, Grundke-Iqbal I (1991) Ubiquitination and abnormal phosphorylation of paired helical filaments in Alzheimer’s disease. Mol Neurobiol 5:399–410CrossRefPubMedGoogle Scholar
  15. Iqbal K, Zaidi T, Bancher C, Grundke-Iqbal I (1994) Alzheimer paired helical filaments. Restoration of the biological activity by dephosphorylation. FEBS Lett 349:104–108CrossRefPubMedGoogle Scholar
  16. Julien C, Marcouiller F, Bretteville A, El Khoury NB, Baillargeon J, Hebert SS, Planel E (2012) Dimethyl sulfoxide induces both direct and indirect tau hyperphosphorylation. PLoS One 7:e40020CrossRefPubMedPubMedCentralGoogle Scholar
  17. LaFerla FM, Tinkle BT, Bieberich CJ, Haudenschild CC, Jay G (1995) The Alzheimer’s A beta peptide induces neurodegeneration and apoptotic cell death in transgenic mice. Nat Genet 9:21–30CrossRefPubMedGoogle Scholar
  18. Lee DH, Goldberg AL (1998) Proteasome inhibitors: valuable new tools for cell biologists. Trends Cell Biol 8:397–403CrossRefPubMedGoogle Scholar
  19. Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2(−Delta Delta C(T)) method. Methods 25:402–408CrossRefPubMedGoogle Scholar
  20. Maas T, Eidenmuller J, Brandt R (2000) Interaction of tau with the neural membrane cortex is regulated by phosphorylation at sites that are modified in paired helical filaments. J Biol Chem 275:15733–15740CrossRefPubMedGoogle Scholar
  21. Maurin H, Lechat B, Borghgraef P, Devijver H, Jaworski T, Van Leuven F (2014) Terminal hypothermic tau. P301L mice have increased tau phosphorylation independently of glycogen synthase kinase 3alpha/beta. Eur J Neurosci 40:2442–2453CrossRefPubMedGoogle Scholar
  22. McLaughlin D, Tsirimonaki E, Vallianatos G, Sakellaridis N, Chatzistamatiou T, Stavropoulos-Gioka C, Tsezou A, Messinis I, Mangoura D (2006) Stable expression of a neuronal dopaminergic progenitor phenotype in cell lines derived from human amniotic fluid cells. J Neurosci Res 83:1190–1200CrossRefPubMedGoogle Scholar
  23. Mosmann T (1983) Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods 65:55–63CrossRefPubMedGoogle Scholar
  24. Petrucelli L, Dickson D, Kehoe K, Taylor J, Snyder H, Grover A, De Lucia M, McGowan E, Lewis J, Prihar G, Kim J, Dillmann WH, Browne SE, Hall A, Voellmy R, Tsuboi Y, Dawson TM, Wolozin B, Hardy J, Hutton M (2004) CHIP and Hsp70 regulate tau ubiquitination, degradation and aggregation. Hum Mol Genet 13:703–714CrossRefPubMedGoogle Scholar
  25. Planel E, Miyasaka T, Launey T, Chui DH, Tanemura K, Sato S, Murayama O, Ishiguro K, Tatebayashi Y, Takashima A (2004) Alterations in glucose metabolism induce hypothermia leading to tau hyperphosphorylation through differential inhibition of kinase and phosphatase activities: implications for Alzheimer’s disease. J Neurosci 24:2401–2411CrossRefPubMedGoogle Scholar
  26. Planel E, Richter KE, Nolan CE, Finley JE, Liu L, Wen Y, Krishnamurthy P, Herman M, Wang L, Schachter JB, Nelson RB, Lau LF, Duff KE (2007) Anesthesia leads to tau hyperphosphorylation through inhibition of phosphatase activity by hypothermia. J Neurosci 27:3090–3097CrossRefPubMedGoogle Scholar
  27. Planel E, Bretteville A, Liu L, Virag L, Du AL, Yu WH, Dickson DW, Whittington RA, Duff KE (2009) Acceleration and persistence of neurofibrillary pathology in a mouse model of tauopathy following anesthesia. FASEB J 23:2595–2604CrossRefPubMedPubMedCentralGoogle Scholar
  28. Pool M, Thiemann J, Bar-Or A, Fournier AE (2008) NeuriteTracer: a novel ImageJ plugin for automated quantification of neurite outgrowth. J Neurosci Methods 168:134–139CrossRefPubMedGoogle Scholar
  29. Poppek D, Keck S, Ermak G, Jung T, Stolzing A, Ullrich O, Davies KJ, Grune T (2006) Phosphorylation inhibits turnover of the tau protein by the proteasome: influence of RCAN1 and oxidative stress. Biochem J 400:511–520CrossRefPubMedPubMedCentralGoogle Scholar
  30. Rojo LE, Alzate-Morales J, Saavedra IN, Davies P, Maccioni RB (2010) Selective interaction of lansoprazole and astemizole with tau polymers: potential new clinical use in diagnosis of Alzheimer’s disease. J Alzheimers Dis 19:573–589PubMedPubMedCentralGoogle Scholar
  31. Ruiz-Leon Y, Pascual A (2003) Induction of tyrosine kinase receptor b by retinoic acid allows brain-derived neurotrophic factor-induced amyloid precursor protein gene expression in human SH-SY5Y neuroblastoma cells. Neuroscience 120:1019–1026CrossRefPubMedGoogle Scholar
  32. Run X, Liang Z, Zhang L, Iqbal K, Grundke-Iqbal I, Gong CX (2009) Anesthesia induces phosphorylation of tau. J Alzheimers Dis 16:619–626PubMedPubMedCentralGoogle Scholar
  33. Santiago FE, Almeida MC, Carrettiero DC (2015) BAG2 is repressed by NF-κB signaling, and its overexpression is sufficient to shift Aß1-42 from neurotrophic to neurotoxic in undifferentiated SH-SY5Y neuroblastoma. J Mol Neurosci. doi: 10.1007/s12031-015-0579-5 PubMedGoogle Scholar
  34. Shimura H, Miura-Shimura Y, Kosik KS (2004a) Binding of tau to heat shock protein 27 leads to decreased concentration of hyperphosphorylated tau and enhanced cell survival. J Biol Chem 279:17957–17962CrossRefPubMedGoogle Scholar
  35. Shimura H, Schwartz D, Gygi SP, Kosik KS (2004b) CHIP-Hsc70 complex ubiquitinates phosphorylated tau and enhances cell survival. J Biol Chem 279:4869–4876CrossRefPubMedGoogle Scholar
  36. Sontag E, Nunbhakdi-Craig V, Lee G, Brandt R, Kamibayashi C, Kuret J, White CL 3rd, Mumby MC, Bloom GS (1999) Molecular interactions among protein phosphatase 2A, tau, and microtubules. Implications for the regulation of tau phosphorylation and the development of tauopathies. J Biol Chem 274:25490–25498CrossRefPubMedGoogle Scholar
  37. Stieler JT, Bullmann T, Kohl F, Toien O, Bruckner MK, Hartig W, Barnes BM, Arendt T (2011) The physiological link between metabolic rate depression and tau phosphorylation in mammalian hibernation. PLoS One 6:e14530CrossRefPubMedPubMedCentralGoogle Scholar
  38. Wang Y, Mandelkow E (2012) Degradation of tau protein by autophagy and proteasomal pathways. Biochem Soc Trans 40:644–652CrossRefPubMedGoogle Scholar
  39. Whittington RA, Papon MA, Chouinard F, Planel E (2010) Hypothermia and Alzheimer’s disease neuropathogenic pathways. Curr Alzheimer Res 7:717–725CrossRefPubMedGoogle Scholar
  40. Whittington RA, Bretteville A, Virag L, Emala CW, Maurin TO, Marcouiller F, Julien C, Petry FR, El-Khoury NB, Morin F, Charron J, Planel E (2013) Anesthesia-induced hypothermia mediates decreased ARC gene and protein expression through ERK/MAPK inactivation. Sci Rep 3:1388CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  • Cesar Augusto Dias de Paula
    • 1
  • Fernando Enrique Santiago
    • 1
  • Adriele Silva Alves de Oliveira
    • 1
  • Fernando Augusto Oliveira
    • 1
    • 3
  • Maria Camila Almeida
    • 1
    • 2
  • Daniel Carneiro Carrettiero
    • 1
    • 2
    Email author
  1. 1.Universidade Federal do ABCSão Bernardo do CampoBrazil
  2. 2.Centro de Ciências Naturais e HumanasUniversidade Federal do ABCSão Bernardo do CampoBrazil
  3. 3.Centro de Matemática, Computação e CogniçãoUniversidade Federal do ABCSão Bernardo do CampoBrazil

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