Continuous MPTP intoxication in the Göttingen minipig results in chronic parkinsonian deficits


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Acta Neurobiologiae Experimentalis

Nencki Institute of Experimental Biology

Polish Neuroscience Society

Subject: Behavioral Sciences , Biomedical Sciences & Nutrition , Life Sciences , Medicine , Neurosciences


ISSN: 0065-1400
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VOLUME 76 , ISSUE 3 (September 2016) > List of articles

Advertisement Continuous MPTP intoxication in the Göttingen minipig results in chronic parkinsonian deficits

Mette Slot Nielsen * / Andreas Nørgaard Glud / Arne Møller / Poul Mogensen / Dirk Bender / Jens Christian Sørensen / Doris Doudet / Carsten Reidies Bjarkam

Keywords : 1-methyl-4-phenyl-1, 2, 3, 6-tetrahydropyridine (MPTP), animal model, behavior, minipig, neuropathology, Parkinson’s disease

Citation Information : Acta Neurobiologiae Experimentalis. Volume 76, Issue 3, Pages 199-211, DOI:

License : (CC BY 4.0)

Published Online: 01-August-2017



Parkinson’s disease (PD) is a common neurodegenerative disorder, resulting from progressive loss of dopaminergic neurons in the substantia nigra pars compacta (SNc). Neuroprotective therapies in PD are still not available, perhaps because animal models do not imitate the chronic and progressive nature of the clinical state of PD. To address this, we performed a feasibility study aimed at establishing a chronic non-primate large animal PD model in Göttingen minipigs based on continuous infusion of the neurotoxin 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP). Twelve female Göttingen minipigs were divided into groups of 2–4 animals and implanted with infusion pumps for continuous intramuscular MPTP delivery of 4–24 mg MPTP/day for 11 weeks. The animals showed parkinsonian symptoms with bradykinesia, rigidity, coordination and chewing difficulties. Symptoms were stable in the 12 and 18 mg MPTP/day groups, whereas the remaining groups showed partial or full behavioral recovery. Digital gait analysis, high performance liquid chromatography (HPLC) measurements and stereological counts of tyrosine hydroxylase-positive (TH+) neurons in the SNc revealed a dose-related decrease in gait velocity, striatal metabolite levels and neuron numbers with increasing doses of MPTP. No neuronal inclusions were observed, but alpha synuclein staining intensified with increased cumulative MPTP dosages. We conclude that this large-animal model of chronic MPTP administration in Göttingen minipigs shows trends of stable parkinsonian deficits at 18 mg MPTP/day in all modalities examined. This PD model shares many of the characteristics seen in patients and, although preliminary, holds considerable promise for future pre-clinical trials of neuroprotective therapies.

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  1. Alvarez-Fischer D, Guerreiro S, Hunot S, Saurini F, Marien M, Sokoloff P, Hirsch EC, Hartmann A, Michelet PP (2008) Modelling Parkinson-like neurodegeneration via osmotic minipump delivery of MPTP and probenecid. J Neurochem 107: 701–711.
  2. Andersen F, Watanabe H, Bjarkam C, Danielsen EH, Cumming P, DaNeX Study Group (2005) Pig brain stereotaxic standard space: mapping of cerebral blood flow normative values and effect of MPTP-lesioning. Brain Res Bull 66: 17–29.
  3. Aziz T (2006) Essential animals. New Sci 191: 20. Ballard PA, Tetrud JW, Langston JW (1985) Permanent human parkinsonism due to 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP): seven cases. Neurology 35: 949–956.
  4. Baxa M, Hruska-Plochan M, Juhas S, Vodicka P, Pavlok A, Juhasova J, Miyanohara A, Nejime T, Klima J, Macakova M, Marsala S, Weiss A, Kubickova S, Musilova P, Vrtel R, Sontag EM, Thompson LM, Schier J, Hansikova H, Howland DS, Cattaneo E, DiFiglia M, Marsala M, Motlik J (2013) A transgenic minipig model of Huntington’s Disease. J Huntingtons Dis 2(1): 47–68.
  5. Bergman H, Wichmann T, DeLong MR (1990) Reversal of experimental parkinsonism by lesions of the subthalamic nucleus. Science 249: 1436–1438.
  6. Betarbet R, Sherer TB, MacKenzie G, Garcia-Osuna M, Panov AV, Greenamyre JT (2000) Chronic systemic pesticide exposure reproduces features of Parkinson’s disease. Nat Neurosci 3: 1301–1306.
  7. Bezard E, Imbert C, Deloire X, Bioulac B, Gross CE (1997) A chronic MPTP model reproducing the slow evolution of Parkinson’s disease: evolution of motor symptoms in the monkey. Brain Res 766: 107–112.
  8. Bjarkam CR, Cancian G, Glud AN, Ettrup KS, Jorgensen RL, Sorensen JC (2009) MRI-guided stereotaxic targeting in pigs based on a stereotaxic localizer box fitted with an isocentric frame and use of SurgiPlan computer planning software. J Neurosci Methods 183: 119–126.
  9. Bjarkam CR, Cancian G, Larsen M, Rosendahl F, Ettrup KS, Zeidler D, Blankholm AD, Østergaard L, Sunde N, Sørensen JC (2004) A MRI-compatible stereotaxic localizer box enables high-precision stereotaxic procedures in pigs. J Neurosci Methods 139: 293–298.
  10. Bjarkam CR, Glud AN, Margolin L, Reinhart K, Franklin R, Deding D, Ettrup KS, Fitting LM, Nielsen MS, Sørensen JC, Cunningham MG (2010) Safety and function of a new clinical intracerebral microinjection instrument for stem cells and therapeutics examined in the Göttingen minipig. Stereotact Funct Neurosurg 88: 56–63.
  11. Bjarkam CR, Jorgensen R, Jensen KN, Sunde NAa, Sørensen JCH (2008a) Deep brain stimulation electrode anchoring using BioGlue ®, a protective electrode covering, and a titanium microplate. J Neurosci Methods 168: 151–155.
  12. Bjarkam CR, Nielsen, MS, Glud AN, Rosendal F, Mogensen P, Bender D, Doudet D, Møller A, Sørensen JC (2008b) Neuromodulation in a minipig MPTP model of Parkinson disease. Br J Neurosurg 22 Suppl 1: S9–S12.
  13. Bjarkam CR, Sørensen JC, Sunde NA, Geneser FA, Ostergaard K (2001) New strategies for the treatment of Parkinson’s disease hold considerable promise for the future management of neurodegenerative disorders. Biogerontology 2: 193–207.
  14. Boulet S, Mounayar S, Poupard A, Bertrand A, Jan C, Pessiglione M, Hirsch EC, Feuerstein C, François C, Féger J, Savasta M, Tremblay L (2008) Behavioral recovery in MPTP-treated monkeys: neurochemical mechanisms studied by intrastriatal microdialysis. J Neurosci 28: 9575–9584.
  15. Bové J, Prou D, Perier C, Przedborski S (2005) Toxin-induced models of Parkinson’s disease. NeuroRx 2: 484–494.
  16. Braak H, Del Tredici K, Rub U, de Vos RA, Jansen Steur EN, Braak E (2003) Staging of brain pathology related to sporadic Parkinson’s disease. Neurobiol Aging 24: 197–211.
  17. Carlsson A (2002) Treatment of Parkinson’s with L-DOPA. The early discovery phase, and a comment on current problems. J Neural Transm (Vienna) 109: 777–787.
  18. Carvey PM, Punati A, Newman MB (2006) Progressive dopamine neuron loss in Parkinson’s disease: the multiple hit hypothesis. Cell Transplant 15: 239–250.
  19. Dall AM, Danielsen EH, Sørensen JC, Andersen F, Møller A, Zimmer J, Gjedde AH, Cumming P, Danish Neuronal Xenografting Group (2002) Quantitative [18F]fluorodopa/PET and histology of fetal mesencephalic dopaminergic grafts to the striatum of MPTP-poisoned minipigs. Cell Transplant 11: 733–746.
  20. Dalmose AL, Bjarkam CR, Djurhuus JC (2005) Stereotactic electrical stimulation of the pontine micturition centre in the pig. BJU Int 95: 886–889.
  21. Dalmose AL, Bjarkam CR, Sørensen JC, Djurhuus JC, Jørgensen TM (2004) Effects of high frequency deep brain stimulation on urine storage and voiding function in conscious minipigs. Neurourol Urodyn 23: 265–272.
  22. Danielsen EH, Cumming P, Andersen F, Bender D, Brevig T, Falborg L, Gee A, Gillings NM, Hansen SB, Hermansen F, Johansen J, Johansen TE, Dahl-Jørgensen A, Jørgensen HA, Meyer M, Munk O, Pedersen EB, Poulsen PH, Rodell AB, Sakoh M, Simonsen CZ, Smith DF, Sørensen JC, Ostergård L, Zimmer J, Gjedde A, Møller A (2000) The DaNeX study of embryonic mesencephalic, dopaminergic tissue grafted to a minipig model of Parkinson’s disease: preliminary findings of effect of MPTP poisoning on striatal dopaminergic markers. Cell Transplant 9: 247–259.
  23. Dolezalova D, Hruska-Plochan M, Bjarkam CR, Sørensen JCH, Cunningham M, Weingarten D, Ciacci JD, Juhas S, Juhasova J, Motlik J, Hefferan MP, Hazel T, Johe K, Carromeu C, Muotri A, Bui J, Strnadel J, Marsala M (2014) Pig models of neurodegenerative disorders: Utilization in cell replacement-based preclinical and efficacy studies. J Comp Neurol 522: 2784–2801.
  24. Doudet DJ, Chan GL, Holden JE, McGeer EG, Aigner TA, Wyatt RJ, Ruth TJ (1998) 6-[18F]Fluoro-L-DOPA PET studies of the turnover of dopamine in MPTP-induced parkinsonism in monkeys. Synapse 29: 225–232.
  25. Doudet DJ, Cornfeldt ML, Honey CR, Schweikert AW, Allen RC (2004) PET imaging of implanted human retinal pigment epithelial cells in the MPTP-induced primate model of Parkinson’s disease. Exp Neurol 189: 361–368.
  26. Emborg M (2004) Evaluation of animal models of Parkinson’s disease for neuroprotective strategies. J Neurosci Methods 139: 121–143.
  27. Emborg ME (2007) Nonhuman primate models of Parkinson’s disease. ILAR J 48: 339–355.
  28. Ettrup KS, Sørensen JC, Bjarkam CR (2010) The anatomy of the Göttingen minipig hypothalamus. J Chem Neuroanat 39: 151–165.
  29. Ettrup KS, Sørensen JC, Rodell A, Alstrup AK, Bjarkam CR (2012) Hypothalamic deep brain stimulation influences autonomic and limbic circuitry involved in the regulation of aggression and cardiocerebrovascular control in the Göttingen minipig. Stereotact Funct Neurosurg 90(5): 281–291.
  30. Ettrup KS, Tornøe J, Sørensen JC, Bjarkam CR (2011) A surgical device for minimally invasive implantation of experimental deep brain stimulation leads in large research animals. J Neurosci Methods 200(1): 41–46.
  31. Félix B, Léger ME, Albe-Fessard D, Marcilloux JC, Rampin O, Laplace JP (1999) Stereotaxic atlas of the pig brain. Brain Res Bull 49: 1–137.
  32. Fjord-Larsen L, Kusk P, Tornøe J, Juliusson B, Torp M, Bjarkam CR, Nielsen MS, Handberg A, Sørensen JC, Wahlberg LU (2010) Long-term delivery of nerve growth factor by encapsulated cell biodelivery in the Göttingen minipig basal forebrain. Mol Ther 18: 2164–2172.
  33. Fornai F, Schlüter OM, Lenzi P, Gesi M, Ruffoli R, Ferrucci M, Lazzeri G, Busceti CL, Pontarelli F, Battaglia G, Pellegrini A, Nicoletti F, Ruggieri S, Paparelli A, Südhof TC (2005) Parkinson-like syndrome induced by continuous MPTP infusion: convergent roles of the ubiquitin-proteasome system and alpha-synuclein. Proc Natl Acad Sci U S A 102: 3413–3418.
  34. Gamache P, Ryan E, Svendsen C, Murayama K, Acworth IN (1993) Simultaneous Measurement of Monoamines, Metabolites and Amino Acids in Brain Tissue and Microdialysis Perfusates. J Chromatogr 614: 213–220.
  35. Gibrat C, Saint-Pierre M, Bousquet M, Lévesque D, Rouillard C, Cicchetti F (2009) Differences between subacute and chronic MPTP mice models: investigation of dopaminergic neuronal degeneration and alpha-synuclein inclusions. J Neurochem 109: 1469–1482.
  36. Giráldez-Pérez RM, Antolín-Vallespín M, Munoz MD, Sánchez-Capelo A (2014) Models of α-synuclein aggregation in Parkinson’s disease. Acta Neuropathol Commun 2: 176.
  37. Glud AN, Hedegaard C, Nielsen MS, Sørensen JC, Bendixen C, Jensen PH, Mogensen PH, Larsen K, Bjarkam CR (2011) Direct MRI-guided stereotaxic viral mediated gene transfer of alpha-synuclein in the Göttingen minipig CNS. Acta Neurobiol Exp (Wars) 71(4): 508–518.
  38. Goodman S, Check E (2002) The great primate debate. Nature 417: 684–687.
  39. Halliday G, Herrero MT, Murphy K, McCann H, Ros-Bernal F, Barcia C, Mori H, Blesa FJ, Obeso JA (2009) No Lewy pathology in monkeys with over 10 years of severe MPTP parkinsonism. Mov Disord 24: 1519–1523.
  40. Jankovic J (2006) An update on the treatment of Parkinson’s disease. Mt Sinai J Med 73: 682–689.
  41. Jenner P (2008) Functional models of Parkinson’s disease: a valuable tool in the development of novel therapies. Ann Neurol 64 Suppl 2: S16–S29.
  42. Jensen KN, Deding D, Sørensen JC, Bjarkam CR (2009) Long-term implantation of deep brain stimulation electrodes in the pontine micturition centre of the Göttingen minipig. Acta Neurochir (Wien) 151: 785–794.
  43. Johnsen EL, Mogensen PH, Sunde NAa, Østergaard K (2009) Improved asymmetry of gait in Parkinson’s disease with DBS: gait and postural instability in Parkinson’s disease treated with bilateral deep brain stimulation in the subthalamic nucleus. Mov Disord 24: 590–597.
  44. Kalia LV, Kalia SK, Lang AE (2015) Disease-modifying strategies for Parkinson’s disease. Mov Disord 30(11): 1442–1450.
  45. Kieburtz K, Ravina B (2007) Why hasn’t neuroprotection worked in Parkinson’s disease?. Nat Clin Pract Neurol 3: 240–241.
  46. Kragh PM, Nielsen AL, Li J, Du Y, Lin L, Schmidt M, Bøgh IB, Holm IE, Jakobsen JE, Johansen MG, Purup S, Bolund L, Vajta G, Jørgensen AL (2009) Hemizygous minipigs produced by random gene insertion and handmade cloning express the Alzheimer’s disease-causing dominant mutation APPsw. Transgenic Res 2009;18: 545–558.
  47. Larsen M, Bjarkam CR, Østergaard K, West MJ, Sørensen JC (2004) The anatomy of the porcine subthalamic nucleus evaluated with immunohistochemistry and design-based stereology. Anat Embryol (Berl) 208: 239–247.
  48. Lind NM, Moustgaard A, Jelsing J, Vajta G, Cumming P, Hansen AK (2007) The use of pigs in neuroscience: modeling brain disorders. Neurosci Biobehav Rev 31: 728–751.
  49. Lindvall O, Wahlberg LU (2008) Encapsulated cell biodelivery of GDNF: a novel clinical strategy for neuroprotection and neuroregeneration in Parkinson’s disease?. Exp Neurol 209: 82–88.
  50. Markey S, Schmuff N (1986) The pharmacology of the parkinsonian syndrome producing neurotoxin MPTP (1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine) and structurally related compounds. Med Res Rev 6: 389–429.
  51. Meredith GE, Totterdell S, Potashkin JA, Surmeier DJ (2008) Modeling PD pathogenesis in mice: advantages of a chronic MPTP protocol. Parkinsonism Relat Disord 14 Suppl 2: S112–S115.
  52. Mikkelsen M, Møller A, Jensen LH, Pedersen A, Harajehi JB, Pakkenberg H (1999) MPTP-induced Parkinsonism in minipigs: A behavioral, biochemical, and histological study. Neurotoxicol Teratol 21: 169–175.
  53. Mogensen PH, Jakobsen J (2001) Cued and non-cued repetitive ballistic movements. A kinematic study in healthy subjects. Acta Neurol Scand 103: 12–19.
  54. Mounayar S, Boulet S, Tandé D, Jan C, Pessiglione M, Hirsch EC, Féger J, Savasta M, François C, Tremblay L (2007) A new model to study compensatory mechanisms in MPTP-treated monkeys exhibiting recovery. Brain 130: 2898–2914.
  55. Nielsen MS, Sørensen JC, Bjarkam CR (2009) The substantia nigra pars compacta of the Göttingen minipig: an anatomical and stereological study. Brain Struct Funct 213: 481–488.
  56. Norgaard Glud A, Hedegaard C, Nielsen MS, Sørensen JC, Bendixen C, Jensen PH, Larsen K, Bjarkam CR (2010) Direct gene transfer in the Gottingen minipig CNS using stereotaxic lentiviral microinjections. Acta Neurobiol Exp (Wars) 70(3): 308–315.
  57. Olanow CW, Kieburtz K, Schapira AHV (2008) Why have we failed to achieve neuroprotection in Parkinson’s disease?. Ann Neurol 64 Suppl 2: S101–S110.
  58. Ostergaard K, Holm IE, Zimmer J (1992) Tyrosine hydroxylase and acetylcholinesterase in the domestic pig mesencephalon: an immunocytochemical and histochemical study. J Comp Neurol 322: 149–166.
  59. Przedborski S, Jackson-Lewis V, Naini AB, Jakowec M, Petzinger G, Miller R, Akram M (2001) The parkinsonian toxin 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP): a technical review of its utility and safety. J Neurochem 76: 1265–1274.
  60. Rosendal F, Chakravarty MM, Sunde N, Rodell A, Jonsdottir KY, Pedersen M, Bjarkam C, Sørensen JC (2010) Defining the intercommissural plane and stereotactic coordinates for the Basal Ganglia in the Göttingen minipig brain. Stereotact Funct Neurosurg 88: 138–146.
  61. Rosendal F, Frandsen J, Chakravarty MM, Bjarkam CR, Pedersen M, Sangill R, Sorensen JC (2009a) New surgical technique reduces the susceptibility artefact at air-tissue interfaces on in vivo cerebral MRI in the Gottingen minipig. Brain Res Bull 80: 403–407.
  62. Rosendal F, Pedersen M, Sangill R, Stødkilde-Jørgensen H, Nielsen MS, Bjarkam CR, Sunde N, Sorensen JC (2009b) MRI protocol for in vivo visualization of the Göttingen minipig brain improves targeting in experimental functional neurosurgery. Brain Res Bull 79: 41–45.
  63. Schmidt N, Ferger B (2001) Neurochemical findings in the MPTP model of Parkinson’s disease. J Neural Transm (Vienna) 108: 1263–1282.
  64. Seniuk NA, Tatton WG, Greenwood CE (1990) Dose-dependent destruction of the coeruleus-cortical and nigral-striatal projections by MPTP. Brain Res 527: 7–20.
  65. Shimoji M, Zhang L, Mandir AS, Dawson VL, Dawson TM (2005) Absence of inclusion body formation in the MPTP mouse model of Parkinson’s disease. Brain Res Mol Brain Res 134: 103–108.
  66. Song DD, Haber SN (2000) Striatal responses to partial dopaminergic lesion: evidence for compensatory sprouting. J Neurosci 20: 5102–5114.
  67. Sossi V, de La Fuente-Fernandez R, Holden JE, Doudet DJ, McKenzie J, Stoessl AJ, Ruth TJ (2002) Increase in dopamine turnover occurs early in Parkinson’s disease: evidence from a new modeling approach to PET 18 F-fluorodopa data. J Cereb Blood Flow Metab 22: 232–239.
  68. Sørensen JC, Bjarkam CR, Danielsen EH, Simonsen CZ, Geneser FA (2000) Oriented sectioning of irregular tissue blocks in relation to computerized scanning modalities: results from the domestic pig brain. J Neurosci Methods 104: 93–98.
  69. Suchowersky O, Gronseth G, Perlmutter J, Reich S, Zesiewicz T, Weiner WJ (2006) Practice Parameter: neuroprotective strategies and alternative therapies for Parkinson disease (an evidence-based review): report of the Quality Standards Subcommittee of the American Academy of Neurology. Neurology 66: 976–982.
  70. Watanabe H, Andersen F, Simonsen CZ, Evans SM, Gjedde A, Cumming P, DaNex Study Group (2001) MR-based statistical atlas of the Göttingen minipig brain. Neuroimage 14: 1089–1096.
  71. Wernersson R, Schierup MH, Jørgensen FG, Gorodkin J, Panitz F, Staerfeldt H-H, Christensen OF, Mailund T, Hornshøj H, Klein A, Wang J, Liu B, Hu S, Dong W, Li W, Wong GKS, Yu J, Wang J, Bendixen C, Fredholm M, Brunak S, Yang H, Bolund L (2005) Pigs in sequence space: a 0.66X coverage pig genome survey based on shotgun sequencing. BMC Genomics 6: 70.
  72. West MJ (1999) Stereological methods for estimating the total number of neurons and synapses: issues of precision and bias. Trends Neurosci 22: 51–61.
  73. Yacoubian TA, Standaert DG (2009) Targets for neuroprotection in Parkinson’s disease. Biochim Biophys Acta 1792: 676–687.
  74. Yoshihara S, Harada K, Ohta S (2000) Metabolism of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) in perfused rat liver: involvement of hepatic aldehyde oxidase as a detoxification enzyme. Drug Metab Dispos 28: 538–543.