Effects of Treadmill Exercise on Alpha-synuclein Mutation and Activated Neurotrophins in Nigrostriatal Region of MPTP-induced Parkinson Models

MPTP 파킨슨 모델의 트레드밀 운동이 알파시누크린 변성과 흑질선조체내 신경성장인자 활성화에 미치는 영향

  • Park, Jae-Sung (Dept. of Physical Education, College of Education, Korea University) ;
  • Kim, Jeong-Hwan (Dept. of Oriental Rehabilitation Medicine, College of Oriental Medicine, Woo-Suk University) ;
  • Yoon, Sung-Jin (Dept. of Physical Education, College of Education, Korea University)
  • 박재성 (고려대학교 사범대학 체육교육과) ;
  • 김정환 (우석대학교 한의과대학 한방재활의학과교실) ;
  • 윤성진 (고려대학교 사범대학 체육교육과)
  • Received : 2009.03.26
  • Accepted : 2009.04.09
  • Published : 2009.04.30


Objectives : Neuronal changes that result from treadmill exercise for patients with Parkinson's disease(PD) have not been well documented, although some clinical and laboratory reports suggest that regular exercise may produce a neuroprotective effect and restore dopaminergic and motor functions. However, it is not clear if the improvements are due to neuronal alterations within the affected nigrostriatal region or result from a more general effect of exercise on affect areas and motivation. In this study, we demonstrate that motorized treadmill exercise improves the neuronal outcomes in rodent models of PD. Methods : We used a chronic mouse model of parkinsonism, which was induced by injecting male C57BL/6 mice with 10 doses(Every 12 hour) of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (30 mg/kg) and probenecid (20 mg/kg) over 5 days. These mice were able to sustain an exercise training program on a motorized rodent treadmill at a speed of 18 m/min, $0^{\circ}$ of inclination, 40 min/day, 5 days/week for 4 weeks. At the end of exercise training, we extracted the brain and compared their neuronal and neurochemical changes with the control(saline and sedentary) mice groups. Synphilin protein is the substance that manifestly reacts with ${\alpha}$-synuclein. In this study, we used Synphilin as a manifest sign of recovery from neurodegeneration. We analyze the brain stems of the substantia nigra and striatum region using the western blotting technique. Results : There were no expression of synphilin in the saline-induced groups. The addition of MPTP(1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine) greatly accelerated synphilin expression which meant an aggregation of ${\alpha}$-synuclein. But, the MPTP-induced treadmill exercise group showed significantly lower expression than the MPTP-induced sedentary group. This means treadmill exercise has a definite effect on the decrease of ${\alpha}$-synuclein aggregation. Conclusions : In this study, our results suggest that treadmill exercise promoted the removal of the aggregation of ${\alpha}$-synuclein, resulting in protection against disease development and blocks the apoptotic process in the chronic parkinsonian mice brain with severe neurodegeneration.


  1. Hornykiewicz O, Kish SJ. Biochemical pathophysiology of Parkinson's disease. In: Advances in neurology(Yahr MD, Bergmann KJ, eds). Cambridge, Ma, USA:Perseus Publishers. 1987;45:19-34.
  2. Knott M. Report of case of parkinsonism treated with proprioceptive facilitation techniques. Physical Ther Rev. 1957;37:229. https://doi.org/10.1093/ptj/37.4.229
  3. Franklyn S, Kohout LJ, Stren GM, Dunning M. Physiotherapy in Parkinson's disease. In: Research progress in Parkinson's disease(Rose FC, Capiledo R, eds). Tunbridge wells: Pittman Medical Ltd. 1981:397-400.
  4. Toole T, Hirsch MA, Forkink A, Lehman DA, Maitland CG. The effects of abalance and strength training program on equilibrium in parkinsonism: a preliminary study. Neurorehabilitation. 2000;14:165-74.
  5. Brasted PJ, Watts C, Torres EM, Robbins TW, Dunnett SB. Behavioural recovery following striatal transplantation: effects of postoperative training and P-zone volume. Exp Brain Res. 1999;128;535-8. https://doi.org/10.1007/s002210050877
  6. Jellinger KA. Post mortem studies in Parkinson's disease-Is it possible to detect brain areas for specific symptoms? J. Neural Transm. Suppl. 1999:56,129.
  7. Petroske E, Meredith GE, Callen S, Totterdell S, Lau YS. Mouse model of Parkinsonism: A comparison between subacute MPTP and chronic MPTP/probenecid treatment. Neuroscience. 2001;106:589-601. https://doi.org/10.1016/S0306-4522(01)00295-0
  8. Trojanowsi JQ, Lee VM. Parkinson's disease and related neurodegenerative synucleinopathies linked to progressive accumulation of synuclein aggregates in brain. Parkinsonism Relat Disord. 2001;7:247-51. https://doi.org/10.1016/S1353-8020(00)00065-1
  9. Conner J, Lauterborn J, Yan Q, Gall C, Varon S. Distribution of brain-dervied neurotrophic factor(BDNF)protein and mRNA in the normal adult rat CNS:evedence for anterograde axonal transport. J Neurosci. 1997;17:2295-313. https://doi.org/10.1523/JNEUROSCI.17-07-02295.1997
  10. Mu JS, Li WP, Yao ZP, Zhou XF. Deprivation of endogenous brain-derived neurotrophic factor results in impairment of spatial learning and memory in adult rats. Brain Res. 1999;835:259-65. https://doi.org/10.1016/S0006-8993(99)01592-9
  11. Kruttgen A, Saxena S, Evangelopoulos ME, Weis J. Neurotrophins and neurodegenerative disease: Receptors stuck in traffic? J Neuropath Exp Neur. 2003;62:340-50. https://doi.org/10.1093/jnen/62.4.340
  12. Colcher A, Simuni T. Clinical manifestations of Parkinson's disease. Med Clin North Am. 1999;83:327-47. https://doi.org/10.1016/S0025-7125(05)70107-3
  13. Batchelor PE, Liberatore GT, Wong JY, Porritt MJ, Frerichs F, Donnan GA, Howells DW. Activated macrophages and microglia induce dopaminergic sprouting in the injured striatum and express brain-derived neurotrophic factor and glial cell line-delived neurotrophic factor. J Neurosci. 1999;19:1708-16. https://doi.org/10.1523/JNEUROSCI.19-05-01708.1999
  14. Lin LF, Doherty DH, Lile JD, Bektesh S, Collins F. GDNF: a glial cellline-derived neurotrophic factor for midbrain dopaminergic neurons. Science. 1993;260:1130-2. https://doi.org/10.1126/science.8493557
  15. Lau YS. Progressive neurodegeneration in the chronic MPRP/probenecid model of Parkinson's disease. In: Parkinson's disease(Ebidi M, Pfeiffer R, eds). CRC Press, Boca Raton, FL; 2005:109-15
  16. Tu PH, Galvin JE, Baba M, Giasson B, Tomita T, Leight S, Nakajo S, Iwatsubo T, Trojanowski JQ, Lee VM. Glial cytoplasmic inclusion in white matter oligodendrocytes of multiple system atrophy brains contain insoluble alpha-synuclein. Ann neurol. 1998;44:415-22. https://doi.org/10.1002/ana.410440324
  17. Tillerson JL, Caudle WM, Reveron ME, Miller GW. Exercise induces behavioral recovery and attenuates neurochemical deficits in rodent models of Parkinson's disease. J Neurosci. 2003;119:899-911. https://doi.org/10.1016/S0306-4522(03)00096-4
  18. Duda JE, Giasson BI, Chen Q, Gur TL, Hurtig HI, Stern MB, Gollomp SM, Ischiropoulos H, Lee VM, Trojanowski JQ. Widespread nitration of pathological inclusions in neurodegenerative synucleinopathies. Am J Pathol. 2000;157:1439-45. https://doi.org/10.1016/S0002-9440(10)64781-5
  19. Iseki E, Marui W, Kosaka K, Akiyama H, Ueda K, Iwatsubo T. Degenerative terminals of the perforant pathway are human alpha-synuclein-immunoreactive in the hippocampus of patients with diffuse Lewy body disease. Neurosci Lett. 1998;258:81-4. https://doi.org/10.1016/S0304-3940(98)00856-8
  20. Lang AE, Lozano AM. Parkinson's disease. First of two parts. N Engl J Med. 1998;339:1044-53. https://doi.org/10.1056/NEJM199810083391506
  21. Tyler WJ, Alonso M, Bramham CR, Pozzo-Miller LD. From acquisition to consolidation: on the role of brain-derived neurotrophic factor signaling in hippocampal-dependent learning. Learn Memory. 2002;9:224-37. https://doi.org/10.1101/lm.51202
  22. Polymeropoulos MH, Lavedan C, Leroy E, Ide SE, Dehejia A, Dutra A, Pike B, Root H, Rubenstein J, Boyer R. Mutation in the a-synuclein gene identified in families with Parkinson's disease. Science. 1997;276:2045-7. https://doi.org/10.1126/science.276.5321.2045
  23. Jackson-Lewis V, Jakowec M, Burke RE, Przedbroski S. Time course and morphology of dopaminergic neuronal death caused by the neurotoxin 1-methyl-4-phenyl-1,2,3,6- tetrahydropyridine. Neurodegeneration. 1995;4:257-69. https://doi.org/10.1016/1055-8330(95)90015-2
  24. Tatton NA, Kish SJ. In situ detection of apoptotic nuclei in the subtantia nigra compacta of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine-treated mice using terminal deoxynucleotidyl transferase labelling and acridine orange staining. J Neurosci. 1997;77:1037-48. https://doi.org/10.1016/S0306-4522(96)00545-3
  25. Saito Y, Kawashima A, Ruberu NN, Fujiwara H, Koyama S, Sawabe M, Arai T, Nagura H, Yamanouchi H, Hasegawa M, Iwatsubo T, Murayama S. Accumulation of phosphorylated alpha-synuclein in aging human brain. J Neuropathol Wxp Neurol. 2003;62:644-54. https://doi.org/10.1093/jnen/62.6.644
  26. Tillerson JL, Cohen AD, Caudle WM, Zigmond MJ, Schallert T, Miller GW. Forced nonuse in unilateral Parkinsonian rats exacerbates injury. J Neurosci. 2002;22:6790-9.
  27. Kumar R, Lozano AM, Kim YJ, Hutchison WD, Sime E, Halket E, Lang AE. Double-blind evaluation of subthalamic nucleus deep brain stimulation in advanced Parkinson's disease. Neurology. 1998;51:850-5. https://doi.org/10.1212/WNL.51.3.850
  28. Li L, Wu W, Lin LF, Lei M, Oppenheim RW, Houenou LJ. Rescue of adult mouse motoneurons from injury-induced cell death by glial cell line-derived neurotrophic factor on hippocampal neurons after traumatic brain injury in rats. J Neurosurg. 2001;95:674-9. https://doi.org/10.3171/jns.2001.95.4.0674
  29. Tomac A, Lindqvist E, Lin LF, Ogren SO, Young D, Hoffer BJ, Olson L. Protection and repair of the nigrostriatal dopaminergic system by GDNF in vivo. Nature. 1995;373:335-9. https://doi.org/10.1038/373335a0
  30. Serra PA, Sciola L, Delogu MR. The neurotoxin 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine induces apoptosis in mouse of nigrostriatal glia. Relevance to nigral neuronal death and striatal neurochemical changes. J Biol Chem. 2002;277:3451-61.
  31. Szekely BC, kosanovich NN, Sheppard W. Adjunctive treatment in Parkinson's disease: physical therapy and comprehensive group therapy. Rehabil Lit. 1982;43:72-6.
  32. Lau YS, Trobough KL, Crampton JM, Wilson JA. Effects of probenecid on striatal dopamine depletion in acute and long-term 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine(MPTP)-treated mice. Gen Phamacol. 1990;21:181-7.
  33. Das K, Chao S, White L, Haines W, Harry G, Tilson H. Differential patterns of nerve growth factor, brain-derived neurotrophic factor and neurotrophin-3 mRNA and protein levels in developing regions of rat brain. J Neurosci. 2001;103:739-61. https://doi.org/10.1016/S0306-4522(01)00011-2
  34. Hashimoto M, Hsu LJ, Sisk A, Xia Y, Takeda A, Sundsmo M, Masliah E. Human recombinant NACP/alpha-synuclein is aggregated and fibrillated in vitro: relevance for Lewy body disease. Brain Res. 1998;799:301-6. https://doi.org/10.1016/S0006-8993(98)00514-9
  35. Jones TA, Schallert T. Use-dependent growth of pyramidal neurons after neocortical damage. J Neurosci. 1994;14:2140-52. https://doi.org/10.1523/JNEUROSCI.14-04-02140.1994
  36. Giasson BI, Duda JE, Murray IV, Chen Q, Souza JM, Hurting HI, Ischiropoulos H, Trojanowski JQ, Lee VM. Oxidative damage linked to neurodegeneration by selective alpha-synuclein nitration in synucleinopathy lesions. Science. 2000;290:985-9. https://doi.org/10.1126/science.290.5493.985
  37. Novikova L, Garris BL, Garris DR, Lau YS. Early signs of neuronal apoptosis in the subtantia nigra pars compacta of the progressive neurodegenerative mouse 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine/probenecid model of Parkinson's disease. Neuroscience. 2006;140:67-76. https://doi.org/10.1016/j.neuroscience.2006.02.007
  38. Murer M, Yan Q, Rainman-Vozari R. Brain-derived neurotrophic factor in the control human brain, and in Alzheimer's disease and Parkinson's disease. Progneurobiol. 2001;63:71-124.
  39. karege F, Perret G, Bondolfi G, Schwald M, Bertschy G, Aubry JM. Decreased serum brain-derived neurotrophic factor levels in major depressed patients. Psychiat Res. 2002;109:143-8. https://doi.org/10.1016/S0165-1781(02)00005-7
  40. 1Lewy FH, Lewandowski M, Abelsdorff G. In Paralysis Agitans. Handbuch der Neurologie. Springer-Verlag, Berlin, Ed. III, S.1912;920-33.
  41. Fisher BE, Petzinger GM, Nixon K, Hogg E, Bremmer S, Meshul CK, Jakowec MW. Exercise-induced behavioral recovery and neuroplasticity in the 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine-leisoned mouse basal ganglia. J Neurosci Res. 2004;7:378-90.
  42. Hurwitz A. The benefit of a home exercise regiment for ambulatory Parkinson's disease patients. J Neurosci. Nurs. 1989;21:180-4. https://doi.org/10.1097/01376517-198906000-00008
  43. Bostantjopoulou S, Katsarou Z, Papadimitriou A, Veletza V, Hatzigeorgiou G, Lees A. Clinical features of parkinsonian patients with the a-synuclein (G209A) mutation. Mov. Disord. 2001;16:1007-13. https://doi.org/10.1002/mds.1221
  44. Hashimoto M, Masliah E. Alpha-synuclein in Lewy body disease and Alzheimer's disease. Brain Pathol. 1999;9:707-29. https://doi.org/10.1111/j.1750-3639.1999.tb00552.x
  45. Spillantini MG, Schmidt ML, Lee VM, Trojanowski JQ, Jakes R, Goedert M. a-Synuclein in Lewy bodies. Nature. 1997;388:839-40. https://doi.org/10.1038/42166
  46. Ivanova T, Beyer C. Pre-and Postnatal expression of brain-derived neurotrophic factor mRNA/protein and tyrosine protein kinase receptorB mRNA in the mouse hippocampus. Neurosci Lett. 2001;307:21-4. https://doi.org/10.1016/S0304-3940(01)01905-X
  47. Fahnestock M, Garzon D, Holsinger RMD, Michalski B. Neurotrophic factors and Alzheimer's disease: are we focusing on the wrong molecule? J Neuro Transm Suppl. 2002;62:241-52.
  48. Sasco AJ, Paffenbarger RS Jr, Gendre I, Wing Al. Therole of physical exercise in the occurrence of Parkinson's disease. Arch neurol. 1992;49:360-5. https://doi.org/10.1001/archneur.1992.00530280040020
  49. Kitagawa H, Hayashi T, Mitsumoto Y, Koga N, Itoyama Y, Abe K. Reduction of ischemic brain injury by topical application of glial cell line-derived neurotrophic factor after permanent middle cerebral artery occlusion in rats. Stroke. 1998;29:1417-22. https://doi.org/10.1161/01.STR.29.7.1417
  50. Adler CH. Nonmotor complications in Parkinson's disease. Mov Disorder 20 Suppl. 2005;11:S23-9.
  51. Bilowit DS. Establishing physical objectives in rehabilitation of patients with Parkinson's disease (gymnasium activities). Physical Ther Rev. 1956;36:176-8. https://doi.org/10.1093/ptj/36.3.176
  52. Palmer SS, Mortimer JA, Webster DD, Bistevins R, Dickinson GL. Exercise therapy for Parkinson's disease. Arch. Phys. Med. Rehab. 1986;67:741-5. https://doi.org/10.1016/0003-9993(86)90007-9
  53. Baba M, Nakajo S, Tu PH, Tomita T, Nakaya K, Lee VM, Trojanowski JQ, Iwatsubo T. Aggregation of alpha-synuclein in Lewy bodies of sporadic Parkinson's disease and dementia with Lewy bodies. Am J Pathol. 1998;152:879-84.
  54. Hirsch EC. Nifrostriatal system plasticity in Parkinson's disease: effect of dopaminergic denervation and treatment. Ann neurol. 2000;47:S115-20.
  55. Fujiwara H, Hasegawa M, Dohmae N, Kawashima A, Masliah E, Goldberg MS, Shen J, Takio K, Iwatsubo T. Alpha-Synuclein is phosphorylated in synucleinopathy lesions. Nat Cell Biol. 2002;4:160-4. https://doi.org/10.1038/ncb748