The Cerebellum

, Volume 8, Issue 3, pp 163–174

Cuprizone Treatment Induces Distinct Demyelination, Astrocytosis, and Microglia Cell Invasion or Proliferation in the Mouse Cerebellum

  • Angela Groebe
  • Tim Clarner
  • Werner Baumgartner
  • Jon Dang
  • Cordian Beyer
  • Markus Kipp


Demyelination of the cerebellum is a well-known phenomenon in human multiple sclerosis (MS). Concordantly, patients with MS frequently developed symptoms deriving from cerebellar lesions, i.e., dysmetria leading to hand dexterity impairment. Important advances in MS research have been made as a direct or indirect consequence of the establishment of adequate animal models. In this study, we used the cuprizone mouse model to investigate cerebellar demyelination in young adult male mice. The myelin status was analyzed by immunohistochemistry for proteolipoprotein and electron microscopy. The expression and presence of oligodendrocyte, astroglial, and microglia markers were supplementary studied. Cuprizone intoxication induced an almost complete demyelination of cerebellar nuclei. Cerebellar cortex regions were not (cortical gray matter) or only marginally (cortical white matter) affected. In addition, the affected areas displayed hypertrophic and hyperplastic astrocytosis accompanied by microglia or macrophage invasion. We conclude that cuprizone-induced demyelination pictures cerebellar deep gray matter involvement but not cerebellar cortex pathology as described for human MS. Behavioral changes after cuprizone described for this animal model may not only result from effects on commissural fiber tracts but also can arise from cerebellar demyelination.


Cuprizone Cerebellum Astrocytes Microglia Oligodendrocyte 



adenomatous polyposis coli protein


cerebellar cortex


cerebellar marrow


central nervous system


bis-cyclohexanone oxaldihydrazone


experimental autoimmune encephalomyelitis


glial fibrillary acidic protein


granular layer


hypoxanthine-guanine phosphoribosyltransferase


ionized calcium binding adaptor molecule




interpositus nucleus


lateral cerebellar nucleus


myelin basic protein


medial cerebellar nucleus


molecular layer


magnet resonance imaging


multiple sclerosis


pyramidal layer




reverse transcription


real time


white matter


  1. 1.
    Noseworthy JH, Lucchinetti C, Rodriguez M, Weinshenker BG (2000) Multiple sclerosis. N Engl J Med 343(13):938–992PubMedCrossRefGoogle Scholar
  2. 2.
    Kidd D, Barkhof F, McConnell R, Algra PR, Allen IV, Revesz T (1999) Cortical lesions in multiple sclerosis. Brain 122(Pt 1):17–26PubMedCrossRefGoogle Scholar
  3. 3.
    Kutzelnigg A, Lassmann H (2006) Cortical demyelination in multiple sclerosis: a substrate for cognitive deficits? J Neurol Sci 245(1–2):123–126PubMedCrossRefGoogle Scholar
  4. 4.
    Pokryszko-Dragan A, Gruszka E, Bilinska M, Dubik-Jezierzanska M (2008) Secondary progressive multiple sclerosis - clinical course and potential predictive factors. Neurol Neurochir Pol 42(1):6–11PubMedGoogle Scholar
  5. 5.
    Kutzelnigg A, Faber-Rod JC, Bauer J, Lucchinetti CF, Sorensen PS, Laursen H et al (2007) Widespread demyelination in the cerebellar cortex in multiple sclerosis. Brain Pathol 17(1):38–44PubMedCrossRefGoogle Scholar
  6. 6.
    Gilmore CP, Donaldson I, Bo L, Owens T, Lowe JS, Evangelou N (2009) Regional variations in the extent and pattern of grey matter demyelination in Multiple Sclerosis: a comparison between the cerebral cortex, cerebellar cortex, deep grey matter nuclei and the spinal cord. J Neurol Neurosurg Psychiatry 80(2):182–187PubMedCrossRefGoogle Scholar
  7. 7.
    Craner MJ, Lo AC, Black JA, Baker D, Newcombe J, Cuzner ML et al (2003) Annexin II/p11 is up-regulated in Purkinje cells in EAE and MS. Neuroreport 14(4):555–558PubMedCrossRefGoogle Scholar
  8. 8.
    Tonra JR (2002) Cerebellar susceptibility to experimental autoimmune encephalomyelitis in SJL/J mice: potential interaction of immunology with vascular anatomy. Cerebellum 1(1):57–68PubMedCrossRefGoogle Scholar
  9. 9.
    Yousry TA, Grossman RI, Filippi M (2000) Assessment of posterior fossa damage in MS using MRI. J Neurol Sci 172(Suppl 1):S50–S53PubMedCrossRefGoogle Scholar
  10. 10.
    Acs P, Kipp M, Norkute A, Johann S, Clarner T, Braun A, Berente Z, Komoly S, Beyer C (2008) 17beta-estradiol and progesterone prevent cuprizone provoked demyelination of corpus callosum in male mice. Glia, in pressGoogle Scholar
  11. 11.
    Franklin RJ, Ffrench-Constant C (2008) Remyelination in the CNS: from biology to therapy. Nat Rev Neurosci 9(11):839–855PubMedCrossRefGoogle Scholar
  12. 12.
    Lassmann H, Bruck W, Lucchinetti C (2001) Heterogeneity of multiple sclerosis pathogenesis: implications for diagnosis and therapy. Trends Mol Med 7(3):115–121PubMedCrossRefGoogle Scholar
  13. 13.
    Franco-Pons N, Torrente M, Colomina MT, Vilella E (2007) Behavioral deficits in the cuprizone-induced murine model of demyelination/remyelination. Toxicol Lett 169(3):205–213PubMedCrossRefGoogle Scholar
  14. 14.
    Liebetanz D, Merkler D (2006) Effects of commissural de- and remyelination on motor skill behaviour in the cuprizone mouse model of multiple sclerosis. Exp Neurol 202(1):217–224PubMedCrossRefGoogle Scholar
  15. 15.
    Matsushima GK, Morell P (2001) The neurotoxicant, cuprizone, as a model to study demyelination and remyelination in the central nervous system. Brain Pathol 11(1):107–116PubMedCrossRefGoogle Scholar
  16. 16.
    Stott SR, Kirik D (2006) Targeted in utero delivery of a retroviral vector for gene transfer in the rodent brain. Eur J Neurosci 24(7):1897–1906PubMedCrossRefGoogle Scholar
  17. 17.
    Cao Q, Xu XM, Devries WH, Enzmann GU, Ping P, Tsoulfas P et al (2005) Functional recovery in traumatic spinal cord injury after transplantation of multineurotrophin-expressing glial-restricted precursor cells. J Neurosci 25(30):6947–6957PubMedCrossRefGoogle Scholar
  18. 18.
    Tekkok SB, Goldberg MP (2001) Ampa/kainate receptor activation mediates hypoxic oligodendrocyte death and axonal injury in cerebral white matter. J Neurosci 21(12):4237–4248PubMedGoogle Scholar
  19. 19.
    Cheung KK, Mok SC, Rezaie P, Chan WY (2008) Dynamic expression of Dab2 in the mouse embryonic central nervous system. BMC Dev Biol 8(1):76PubMedCrossRefGoogle Scholar
  20. 20.
    Wells JE, Biernaskie J, Szymanska A, Larsen PH, Yong VW, Corbett D (2005) Matrix metalloproteinase (MMP)-12 expression has a negative impact on sensorimotor function following intracerebral haemorrhage in mice. Eur J Neurosci 21(1):187–196PubMedCrossRefGoogle Scholar
  21. 21.
    Kipp M, Norkute A, Johann S, Lorenz L, Braun A, Hieble A et al (2008) Brain-region-specific astroglial responses in vitro after LPS exposure. J Mol Neurosci 35(2):235–243PubMedCrossRefGoogle Scholar
  22. 22.
    Kipp M, Karakaya S, Johann S, Kampmann E, Mey J, Beyer C (2007) Oestrogen and progesterone reduce lipopolysaccharide-induced expression of tumour necrosis factor-alpha and interleukin-18 in midbrain astrocytes. J Neuroendocrinol 19(10):819–822PubMedCrossRefGoogle Scholar
  23. 23.
    Alusi SH, Worthington J, Glickman S, Bain PG (2001) A study of tremor in multiple sclerosis. Brain 124(Pt 4):720–730PubMedCrossRefGoogle Scholar
  24. 24.
    Ito M (2006) Cerebellar circuitry as a neuronal machine. Prog Neurobiol 78(3-5):272–303PubMedCrossRefGoogle Scholar
  25. 25.
    Schwarz C, Thier P (1999) Binding of signals relevant for action: towards a hypothesis of the functional role of the pontine nuclei. Trends Neurosci 22(10):443–451PubMedCrossRefGoogle Scholar
  26. 26.
    Mitosek-Szewczyk K, Sulkowski G, Stelmasiak Z, Struzynska L (2008) Expression of glutamate transporters GLT-1 and GLAST in different regions of rat brain during the course of experimental autoimmune encephalomyelitis. Neuroscience 155(1):45–52PubMedCrossRefGoogle Scholar
  27. 27.
    Kis B, Rumberg B, Berlit P (2008) Clinical characteristics of patients with late-onset multiple sclerosis. J Neurol 255(5):697–702PubMedCrossRefGoogle Scholar
  28. 28.
    Dubois-Dalcq M, Ffrench-Constant C, Franklin RJ (2005) Enhancing central nervous system remyelination in multiple sclerosis. Neuron 48(1):9–12PubMedCrossRefGoogle Scholar
  29. 29.
    Torkildsen O, Brunborg LA, Myhr KM, Bo L (2008) The cuprizone model for demyelination. Acta Neurol Scand Suppl 188:72–76PubMedCrossRefGoogle Scholar
  30. 30.
    Skripuletz T, Lindner M, Kotsiari A, Garde N, Fokuhl J, Linsmeier F et al (2008) Cortical demyelination is prominent in the murine cuprizone model and is strain-dependent. Am J Pathol 172(4):1053–1061PubMedCrossRefGoogle Scholar
  31. 31.
    Norkute A, Hieble A, Braun A, Johann S, Clarner T, Baumgartner W, Beyer C, Kipp M (2008) Cuprizone treatment induces demyelination and astrocytosis in the mouse hippocampus. J Neurosci Res 19Google Scholar
  32. 32.
    Kleim JA, Pipitone MA, Czerlanis C, Greenough WT (1998) Structural stability within the lateral cerebellar nucleus of the rat following complex motor learning. Neurobiol Learn Mem 69(3):290–306PubMedCrossRefGoogle Scholar
  33. 33.
    Alvina K, Walter JT, Kohn A, Ellis-Davies G, Khodakhah K (2008) Questioning the role of rebound firing in the cerebellum. Nat Neurosci 11(11):1256–1258PubMedCrossRefGoogle Scholar
  34. 34.
    Sanchez-Campusano R, Gruart A, Delgado-Garcia JM (2007) The cerebellar interpositus nucleus and the dynamic control of learned motor responses. J Neurosci 27(25):6620–6632PubMedCrossRefGoogle Scholar
  35. 35.
    Pu YM, Wang JJ, Wang T, Yu QX (1995) Cerebellar interpositus nucleus modulates neuronal activity of lateral hypothalamic area. Neuroreport 6(7):985–988PubMedCrossRefGoogle Scholar
  36. 36.
    Aschoff JC, Conrad B, Kornhuber HH (1974) Acquired pendular nystagmus with oscillopsia in multiple sclerosis: a sign of cerebellar nuclei disease. J Neurol Neurosurg Psychiatry 37(5):570–577PubMedCrossRefGoogle Scholar
  37. 37.
    Tjoa CW, Benedict RH, Weinstock-Guttman B, Fabiano AJ, Bakshi R (2005) MRI T2 hypointensity of the dentate nucleus is related to ambulatory impairment in multiple sclerosis. J Neurol Sci 234(1–2):17–24PubMedCrossRefGoogle Scholar
  38. 38.
    Li Y, Chiaravalloti ND, Hillary FG, Deluca J, Liu WC, Kalnin AJ et al (2004) Differential cerebellar activation on functional magnetic resonance imaging during working memory performance in persons with multiple sclerosis. Arch Phys Med Rehabil 85(4):635–639PubMedCrossRefGoogle Scholar
  39. 39.
    Blakemore WF, Franklin RJ (2008) Remyelination in experimental models of toxin-induced demyelination. Curr Top Microbiol Immunol 318:193–212PubMedCrossRefGoogle Scholar
  40. 40.
    Oleszak EL, Chang JR, Friedman H, Katsetos CD, Platsoucas CD (2004) Theiler’s virus infection: a model for multiple sclerosis. Clin Microbiol Rev 17(1):174–207PubMedCrossRefGoogle Scholar
  41. 41.
    Lucchinetti C, Bruck W, Parisi J, Scheithauer B, Rodriguez M, Lassmann H (2000) Heterogeneity of multiple sclerosis lesions: implications for the pathogenesis of demyelination. Ann Neurol 47(6):707–717PubMedCrossRefGoogle Scholar
  42. 42.
    Barnett MH, Prineas JW (2004) Relapsing and remitting multiple sclerosis: pathology of the newly forming lesion. Ann Neurol 55(4):458–468PubMedCrossRefGoogle Scholar
  43. 43.
    Ludwin SK, Johnson ES (1981) Evidence for a “dying-back” gliopathy in demyelinating disease. Ann Neurol 9(3):301–305PubMedCrossRefGoogle Scholar
  44. 44.
    Komoly S (2005) Experimental demyelination caused by primary oligodendrocyte dystrophy. Regional distribution of the lesions in the nervous system of mice [corrected]. Ideggyogy Sz 58(1–2):40–43PubMedGoogle Scholar
  45. 45.
    Karakaya S, Kipp M, Beyer C (2007) Oestrogen regulates the expression and function of dopamine transporters in astrocytes of the nigrostriatal system. J Neuroendocrinol 19(9):682–690PubMedCrossRefGoogle Scholar
  46. 46.
    Pawlak J, Brito V, Kuppers E, Beyer C (2005) Regulation of glutamate transporter GLAST and GLT-1 expression in astrocytes by estrogen. Brain Res Mol Brain Res 138(1):1–7PubMedCrossRefGoogle Scholar
  47. 47.
    Lassmann H (2008) Models of multiple sclerosis: new insights into pathophysiology and repair. Curr Opin Neurol 21(3):242–247PubMedCrossRefGoogle Scholar
  48. 48.
    Keegan M, Konig F, McClelland R, Bruck W, Morales Y, Bitsch A et al (2005) Relation between humoral pathological changes in multiple sclerosis and response to therapeutic plasma exchange. Lancet 366(9485):579–582PubMedCrossRefGoogle Scholar
  49. 49.
    Williams A, Piaton G, Lubetzki C (2007) Astrocytes–friends or foes in multiple sclerosis. Glia 55(13):1300–1312PubMedCrossRefGoogle Scholar
  50. 50.
    Johann S, Kampmann E, Denecke B, Arnold S, Kipp M, Mey J et al (2008) Expression of enzymes involved in the prostanoid metabolism by cortical astrocytes after LPS-induced inflammation. J Mol Neurosci 34(2):177–185PubMedCrossRefGoogle Scholar
  51. 51.
    Komoly S, Hudson LD, Webster HD, Bondy CA (1992) Insulin-like growth factor I gene expression is induced in astrocytes during experimental demyelination. Proc Natl Acad Sci USA 89(5):1894–1898PubMedCrossRefGoogle Scholar
  52. 52.
    Mason JL, Ye P, Suzuki K, D’Ercole AJ, Matsushima GK (2000) Insulin-like growth factor-1 inhibits mature oligodendrocyte apoptosis during primary demyelination. J Neurosci 20(15):5703–5708PubMedGoogle Scholar
  53. 53.
    McMahon EJ, Suzuki K, Matsushima GK (2002) Peripheral macrophage recruitment in cuprizone-induced CNS demyelination despite an intact blood-brain barrier. J Neuroimmunol 130(1–2):32–45PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2009

Authors and Affiliations

  • Angela Groebe
    • 1
  • Tim Clarner
    • 1
  • Werner Baumgartner
    • 2
  • Jon Dang
    • 1
  • Cordian Beyer
    • 1
  • Markus Kipp
    • 1
  1. 1.Faculty of Medicine, Institute of NeuroanatomyRWTH Aachen UniversityAachenGermany
  2. 2.Department of Cellular Neurobionics, Institute of ZoologyRWTH Aachen UniversityAachenGermany

Personalised recommendations