Skip to main content
SpringerLink
Log in

An initial top-down proteomic analysis of the standard cuprizone mouse model of multiple sclerosis

Journal of Chemical Biology

Abstract

An initial proteomic analysis of the cuprizone mouse model to characterise the breadth of toxicity by assessing cortex, skeletal muscle, spleen and peripheral blood mononuclear cells. Cuprizone treated vs. control mice for an initial characterisation. Select tissues from each group were pooled, analysed in triplicate using two-dimensional gel electrophoresis (2DE) and deep imaging and altered protein species identified using liquid chromatography tandem mass spectrometry (LC/MS/MS). Forty-three proteins were found to be uniquely detectable or undetectable in the cuprizone treatment group across the tissues analysed. Protein species identified in the cortex may potentially be linked to axonal damage in this model, and those in the spleen and peripheral blood mononuclear cells to the minimal peripheral immune cell infiltration into the central nervous system during cuprizone mediated demyelination. Primary oligodendrocytosis has been observed in type III lesions in multiple sclerosis. However, the underlying mechanisms are poorly understood. Cuprizone treatment results in oligodendrocyte apoptosis and secondary demyelination. This initial analysis identified proteins likely related to axonal damage; these may link primary oligodendrocytosis and secondary axonal damage. Furthermore, this appears to be the first study of the cuprizone model to also identify alterations in the proteomes of skeletal muscle, spleen and peripheral blood mononuclear cells. Notably, protein disulphide isomerase was not detected in the cuprizone cohort; its absence has been linked to reduced major histocompatibility class I assembly and reduced antigen presentation. Overall, the results suggest that, like experimental autoimmune encephalomyelitis, results from the standard cuprizone model should be carefully considered relative to clinical multiple sclerosis.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price includes VAT (France)

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Abbreviations

2DE:

Two-dimensional gel electrophoresis

AD:

Alzheimer’s disease

AFD:

Automated frozen disruption

CC:

Corpus callosum

cCBB:

Colloidal Coomassie Brilliant Blue

EAE:

Experimental autoimmune encephalomyelitis

LC/MS/MS:

Liquid chromatography tandem mass spectrometry

MHC:

Major histocompatibility complex

MP:

Membrane protein

MS:

Multiple sclerosis

OCT:

Optimum cutting temperature compound

PBMCs:

Peripheral blood mononuclear cells

PDI:

Protein disulphide isomerase

RT:

Room temperature

SP:

Soluble protein

References

  1. Nakahara J, Maeda M, Aiso S, Suzuki N (2012) Current concepts in multiple sclerosis: autoimmunity versus oligodendrogliopathy. Clin Rev Allerg Immunol 42:26–34

    Article  CAS  Google Scholar 

  2. Partridge MA, Myers SJ, Gopinath S, Coorssen, JR (2015) Proteomics of a conundrum: thoughts on addressing the aetiology versus progression of multiple sclerosis. Prot Clin Appl. doi:10.1002/prca.201400141

  3. Prineas JW, Parratt JD (2012) Oligodendrocytes and the early multiple sclerosis lesion. Ann Neurol 72:18–31

    Article  Google Scholar 

  4. Stys P, Zamponi G, van Minnen J, Geurts J (2012) Will the real multiple sclerosis please stand up? Nat Rev Neurosci 13:507–514

    Article  CAS  Google Scholar 

  5. Barnett M, Prineas J (2004) Relapsing and remitting multiple sclerosis: pathology of the newly forming lesion. Ann Neurol 55:458–468

    Article  Google Scholar 

  6. Kipp M, Clarner T, Dang J, Copray S, Beyer C (2009) The cuprizone animal model: new insights into an old story. Acta Neuropathol 118:723–736

    Article  Google Scholar 

  7. Matsushima GK, Morell P (2001) The neurotoxicant, cuprizone, as a model to study demyelination and remyelination in the central nervous system. Brain Pathol (Zurich, Switz) 11:107–116

    Article  CAS  Google Scholar 

  8. Kalman B, Laitinen K, Komoly S (2007) The involvement of mitochondria in the pathogenesis of multiple sclerosis. J Neuroimmunol 188:1–12

    Article  CAS  Google Scholar 

  9. Wright E, Prasad K, Padula M, Coorssen J (2014) Deep imaging: how much of the proteome does current top-down technology already resolve? PLoS ONE 9(1):e86058

    Article  Google Scholar 

  10. Wright EP, Partridge MA, Padula MP, Gauci VJ, Malladi CS, Coorssen JR (2014) Top-down proteomics: enhancing 2D gel electrophoresis from tissue processing to high-sensitivity protein detection. Proteomics 14:872–889

    Article  CAS  Google Scholar 

  11. Clarner T et al (2012) Myelin debris regulates inflammatory responses in an experimental demyelination animal model and multiple sclerosis lesions. Glia 60:1468–1480

    Article  Google Scholar 

  12. Taylor L, Gilmore W, Ting J, Matsushima G (2010) Cuprizone induces similar demyelination in male and female C57BL/6 mice and results in disruption of the estrous cycle. J Neurosci Res 88:391–402

    Article  CAS  Google Scholar 

  13. Ishibashi T, Dakin K, Stevens B, Lee P, Kozlov S, Stewart C, Fields D (2006) Astrocytes promote myelination in response to electrical impulses. Neuron 49:823–832

    Article  CAS  Google Scholar 

  14. Butt H, Coorssen J (2005) Postfractionation for enhanced proteomic analyses: routine electrophoretic methods increase the resolution of standard 2D-PAGE. J Proteome Res 4:982–991

    Article  CAS  Google Scholar 

  15. Butt H, Coorssen J (2006) Pre-extraction sample handling by automated frozen disruption significantly improves subsequent proteomic analyses. J Proteome Res 5:437–448

    Article  CAS  Google Scholar 

  16. Butt H, Lee M, Pirshahid A, Backlund P, Wood S, Coorssen J (2006) An initial proteomic analysis of human preterm labor: placental membranes. J Proteome Res 5:3161–3172

    Article  Google Scholar 

  17. Butt H, Pfeifer T, Delaney A, Grigliatti T, Tetzlaff W, Coorssen J (2007) Enabling coupled quantitative genomics and proteomics analyses from rat spinal cord samples. Mol Cell Proteom: MCP 6:1574–1588

    Article  CAS  Google Scholar 

  18. Churchward M, Butt H, Lang J, Hsu K, Coorssen J (2005) Enhanced detergent extraction for analysis of membrane proteomes by two-dimensional gel electrophoresis. Proteome Sci 3:5–15

    Article  Google Scholar 

  19. Gauci V, Padula M, Coorssen J (2013) Coomassie blue staining for high sensitivity gel-based proteomics. J Proteome 90:96–106

    Article  CAS  Google Scholar 

  20. Harris L, Churchward M, Butt H, Coorssen J (2007) Assessing detection methods for gel-based proteomic analyses. J Proteome Res 6:1418–1425

    Article  CAS  Google Scholar 

  21. Werner S, Saha J, Broderick C, Zhen E, Higgs R, Duffin K, Smith R (2010) Proteomic analysis of demyelinated and remyelinating brain tissue following dietary cuprizone administration. J Mol Neurosci: MN 42:210–225

    Article  CAS  Google Scholar 

  22. Oliveira B, Coorssen J, Martins-de-Souza D (2014) 2DE: the phoenix of proteomics. J Proteome 104:140–150

    Article  CAS  Google Scholar 

  23. Wu C, Yates J (2003) The application of mass spectrometry to membrane proteomics. Nat Biotechnol 21:262–267

    Article  CAS  Google Scholar 

  24. Irvine KA, Blakemore WF (2006) Age increases axon loss associated with primary demyelination in cuprizone-induced demyelination in C57BL/6 mice. J Neuroimmunol 175:69–76

    Article  CAS  Google Scholar 

  25. Lindner M, Fokuhl J, Linsmeier F, Trebst C, Stangel M (2009) Chronic toxic demyelination in the central nervous system leads to axonal damage despite remyelination. Neurosci Lett 453:120–125

    Article  CAS  Google Scholar 

  26. Rodriguez M (2003) A function of myelin is to protect axons from subsequent injury: implications for deficits in multiple sclerosis. Brain: J Neurol 126:751–752

    Article  Google Scholar 

  27. Mahad D et al (2009) Mitochondrial changes within axons in multiple sclerosis. Brain: J Neurol 132:1161–1174

    Article  Google Scholar 

  28. Suzuki K (1969) Giant hepatic mitochondria: production in mice fed with cuprizone. Science 163:81–82

    Article  CAS  Google Scholar 

  29. Korshunova I, Caroni P, Kolkova K, Berezin V, Bock E, Walmod P (2008) Characterization of BASP1-mediated neurite outgrowth. J Neurosci Res 86:2201–2213

    Article  CAS  Google Scholar 

  30. Braunewell K-H, Klein-Szanto A, Szanto AK (2009) Visinin-like proteins (VSNLs): interaction partners and emerging functions in signal transduction of a subfamily of neuronal Ca2+-sensor proteins. Cell Tissue Res 335:301–316

    Article  CAS  Google Scholar 

  31. Schnurra I, Bernstein HG, Riederer P, Braunewell KH (2001) The neuronal calcium sensor protein VILIP-1 is associated with amyloid plaques and extracellular tangles in Alzheimer’s disease and promotes cell death and tau phosphorylation in vitro: a link between calcium sensors and Alzheimer’s disease? Neurobiol Dis 8:900–909

    Article  CAS  Google Scholar 

  32. Jaiswal MK (2013) Calcium, mitochondria, and the pathogenesis of ALS: the good, the bad, and the ugly. Front Cell Neurosci 7:199

    Article  Google Scholar 

  33. Zündorf G, Reiser G (2011) Calcium dysregulation and homeostasis of neural calcium in the molecular mechanisms of neurodegenerative diseases provide multiple targets for neuroprotection. Antioxid Redox Signal 14:1275–1288

    Article  Google Scholar 

  34. Emerson MR, Biswas S, LeVine SM (2001) Cuprizone and piperonyl butoxide, proposed inhibitors of T-cell function, attenuate experimental allergic encephalomyelitis in SJL mice. J Neuroimmunol 119:205–213

    Article  CAS  Google Scholar 

  35. Hopkins RG, Failla ML (1997) Copper deficiency reduces interleukin-2 (IL-2) production and IL-2 mRNA in human T-lymphocytes. J Nutr 127:257–262

    CAS  Google Scholar 

  36. Maña P, Fordham S, Staykova M, Correcha M, Silva D, Willenborg D, Liñares D (2009) Demyelination caused by the copper chelator cuprizone halts T cell mediated autoimmune neuroinflammation. J Neuroimmunol 210:13–21

    Article  Google Scholar 

  37. Bronte V, Zanovello P (2005) Regulation of immune responses by L-arginine metabolism. Nat Rev Immunol 5:641–654

    Article  CAS  Google Scholar 

  38. Moliné-Velázquez V, Cuervo H, Vila-Del Sol V, Ortega MC, Clemente D, de Castro F (2011) Myeloid-derived suppressor cells limit the inflammation by promoting T lymphocyte apoptosis in the spinal cord of a murine model of multiple sclerosis. Brain Pathol (Zurich, Switz) 21:678–691

    Article  Google Scholar 

  39. Movahedi K et al (2008) Identification of discrete tumor-induced myeloid-derived suppressor cell subpopulations with distinct T cell-suppressive activity. Blood 111:4233–4244

    Article  CAS  Google Scholar 

  40. Grune T, Reinheckel T, Li R, North J, Davies K (2002) Proteasome-dependent turnover of protein disulfide isomerase in oxidatively stressed cells. Arch Biochem Biophys 397:407–413

    Article  CAS  Google Scholar 

  41. Kang K, Park B, Oh C, Cho K, Ahn K (2009) A role for protein disulfide isomerase in the early folding and assembly of MHC class I molecules. Antioxid Redox Signal 11:2553–2561

    Article  CAS  Google Scholar 

  42. Critchley D (2009) Biochemical and structural properties of the integrin-associated cytoskeletal protein talin. Ann Rev Biophys 38:235–254

    Article  CAS  Google Scholar 

  43. Leuker CE, Labow M, Müller W, Wagner N (2001) Neonatally induced inactivation of the vascular cell adhesion molecule 1 gene impairs B cell localization and T cell-dependent humoral immune response. J Exp Med 193:755–768

    Article  CAS  Google Scholar 

  44. Rott LS, Briskin MJ, Butcher EC (2000) Expression of alpha4beta7 and E-selectin ligand by circulating memory B cells: implications for targeted trafficking to mucosal and systemic sites. J Leukoc Biol 68:807–814

    CAS  Google Scholar 

  45. Manevich-Mendelson E et al (2010) Talin 1 is required for integrin-dependent B lymphocyte homing to lymph nodes and the bone marrow but not for follicular B-cell maturation in the spleen. Blood 116:5907–5918

    Article  CAS  Google Scholar 

Download references

Acknowledgments

The authors are grateful to the Rotary Club of Narellan for initiating and funding the Narellan-UWS MS Research Project. MAP acknowledges the UWS Molecular Medicine Research Group and the School of Medicine for PhD scholarship support provided in complement to the funding from Rotary. The authors thank Ashleigh Deschamps and the School of Medicine Animal Facility staff for outstanding assistance, and acknowledge use of the UWS Mass Spectrometry Facility.

Conflict of interest

The authors declare no conflict of interest.

Authors’ contributions

MAP was involved in study design, data acquisition, data analysis and manuscript drafting. SG was involved in manuscript drafting. SJM was involved in study design and manuscript drafting. JRC was involved in study design, data analysis and manuscript drafting. All authors read and approved the final manuscript.

Author information

Authors and Affiliations

  1. Department of Molecular Physiology, School of Medicine, University of Western Sydney, Penrith, NSW, Australia

    Melissa A. Partridge & Jens R Coorssen

  2. Molecular Medicine Research Group, School of Medicine, University of Western Sydney, Penrith, NSW, Australia

    Melissa A. Partridge, Sumana Gopinath, Simon J. Myers & Jens R Coorssen

  3. Neuro-Cell Biology Laboratory, School of Science and Health, University of Western Sydney, Penrith, NSW, Australia

    Simon J. Myers

  4. Department of Neurology, Campbelltown Hospital, Campbelltown, NSW, Australia

    Sumana Gopinath

Authors
  1. Melissa A. Partridge
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Sumana Gopinath
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Simon J. Myers
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Jens R Coorssen
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Jens R Coorssen.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Partridge, M.A., Gopinath, S., Myers, S.J. et al. An initial top-down proteomic analysis of the standard cuprizone mouse model of multiple sclerosis. J Chem Biol 9, 9–18 (2016). https://doi.org/10.1007/s12154-015-0138-0

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s12154-015-0138-0

Keywords

search

Navigation