Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Research Article
  • Published:

Delayed delivery of AAV-GDNF prevents nigral neurodegeneration and promotes functional recovery in a rat model of Parkinson's disease

Gene Therapy volume 9, pages 381–389 (2002)Cite this article

Abstract

Glial cell line-derived neurotrophic factor (GDNF) is a strong candidate agent in the neuroprotective treatment of Parkinson's disease (PD). We investigated whether adeno-associated viral (AAV) vector-mediated delivery of a GDNF gene in a delayed manner could prevent progressive degeneration of dopaminergic (DA) neurons, while preserving a functional nigrostriatal pathway. Four weeks after a unilateral intrastriatal injection of 6-hydroxydopamine (6-OHDA), rats received injection of AAV vectors expressing GDNF tagged with FLAG peptide (AAV-GDNFflag) or β-galactosidase (AAV-LacZ) into the lesioned striatum. Immunostaining for FLAG demonstrated retrograde transport of GDNFflag to the substantia nigra (SN). The density of tyrosine hydroxylase (TH)-positive DA fibers in the striatum and the number of TH-positive or cholera toxin subunit B (CTB, neuronal tracer)-labeled neurons in the SN were significantly greater in the AAV-GDNFflag group than in the AAV-LacZ group. Dopamine levels and those of its metabolites in the striatum were remarkably higher in the AAV-GDNFflag group compared with the control group. Consistent with anatomical and biochemical changes, significant behavioral recovery was observed from 4–20 weeks following AAV-GDNFflag injection. These data indicate that a delayed delivery of GDNF gene using AAV vector is efficacious even 4 weeks after the onset of progressive degeneration in a rat model of PD.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
Figure 6
Figure 7
Figure 8

Similar content being viewed by others

References

  1. Dunnett S.B., Bjorklund A. . Prospects for new restorative and neuroprotective treatments in Parkinson's disease Nature 1999 399: A32 A32

    Article  CAS  Google Scholar 

  2. Bjorklund A. et al. Towards a neuroprotective gene therapy for Parkinson's disease: use of adenovirus, AAV and lentivirus vectors for gene transfer of GDNF to the nigrostriatal system in the rat Parkinson model Brain Res 2000 886: 82 82

    Article  CAS  Google Scholar 

  3. Bohn M.C. . Parkinson's disease: a neurodegenerative disease particularly amenable to gene therapy Mol Ther 2000 1: 494 494

    Article  CAS  Google Scholar 

  4. Ozawa K. et al. Gene therapy of Parkinson's disease using adeno-associated virus (AAV) vectors J Neural Transm Suppl 2000 58: 181 181

    Google Scholar 

  5. Choi-Lundberg D.L. et al. Dopaminergic neurons protected from degeneration by GDNF gene therapy Science 1997 275: 838 838

    Article  CAS  Google Scholar 

  6. Mandel R.J., Spratt S.K., Snyder R.O., Leff S.E. . Midbrain injection of recombinant adeno-associated virus encoding rat glial cell line-derived neurotrophic factor protects nigral neurons in a progressive 6-hydroxydopamine-induced degeneration model of Parkinson's disease in rats Proc Natl Acad Sci USA 1997 94: 14083 14083

    Article  CAS  Google Scholar 

  7. Bilang-Bleuel A. et al. Intrastriatal injection of an adenoviral vector expressing glial cell line-derived neurotrophic factor prevents dopaminergic neuron degeneration and behavioral impairment in a rat model of Parkinson disease Proc Natl Acad Sci USA 1997 94: 8818 8818

    Article  CAS  Google Scholar 

  8. Choi-Lundberg D.L. et al. Behavioral and cellular protection of rat dopaminergic neurons by an adenoviral vector encoding glial cell line-derived neurotrophic factor Exp Neurol 1998 154: 261 261

    Article  CAS  Google Scholar 

  9. Mandel R.J., Snyder R.O., Leff S.E. . Recombinant adeno-associated viral vector-mediated glial cell line-derived neurotrophic factor gene transfer protects nigral dopamine neurons after onset of progressive degeneration in a rat model of Parkinson's disease Exp Neurol 1999 160: 205 205

    Article  CAS  Google Scholar 

  10. Kirik D., Rosenblad C., Bjorklund A., Mandel R.J. . Long-term rAAV-mediated gene transfer of GDNF in the rat Parkinson's model: intrastriatal but not intranigral transduction promotes functional regeneration in the lesioned nigrostriatal system J Neurosci 2000 20: 4686 4686

    Article  CAS  Google Scholar 

  11. Bensadoun J.C. et al. Lentiviral vectors as a gene delivery system in the mouse midbrain: cellular and behavioral improvements in a 6-OHDA model of Parkinson's disease using GDNF Exp Neurol 2000 164: 15 15

    Article  CAS  Google Scholar 

  12. Kordower J.H. et al. Neurodegeneration prevented by lentiviral vector delivery of GDNF in primate models of Parkinson's disease Science 2000 290: 767 767

    Article  CAS  Google Scholar 

  13. Leman S., Viltart O., Sequeira H. . Double immunocytochemistry for the detection of Fos protein in retrogradely identified neurons using cholera toxin B subunit Brain Res Brain Res Protoc 2000 5: 298 298

    Article  CAS  Google Scholar 

  14. Monahan P.E., Samulski R.J. . AAV vectors: is clinical success on the horizon? Gene Therapy 2000 7: 24 24

    Article  CAS  Google Scholar 

  15. Bankiewicz K.S. et al. Convection-enhanced delivery of AAV vector in parkinsonian monkeys; in vivo detection of gene expression and restoration of dopaminergic function using pro-drug approach Exp Neurol 2000 164: 2 2

    Article  CAS  Google Scholar 

  16. Shen Y. . Triple transduction with adeno-associated virus vectors expressing tyrosine hydroxylase aromatic-L-amino-acid decarboxylase and GTP cyclohydrolase I for gene therapy of Parkinson's disease Hum Gene Ther 2000 11: 1509 1509

    Article  CAS  Google Scholar 

  17. Sauer H., Oertel W.H. . Progressive degeneration of nigrostriatal dopamine neurons following intrastriatal terminal lesions with 6-hydroxydopamine: a combined retrograde tracing and immunocytochemical study in the rat Neuroscience 1994 59: 401 401

    Article  CAS  Google Scholar 

  18. Hudson J.L. et al. Correlation of apomorphine- and amphetamine-induced turning with nigrostriatal dopamine content in unilateral 6-hydroxydopamine lesioned rats Brain Res 1993 626: 167 167

    Article  CAS  Google Scholar 

  19. Javoy-Agid F. et al. Decreased tyrosine hydroxylase messenger RNA in the surviving dopamine neurons of the substantia nigra in Parkinson's disease: an in situ hybridization study Neuroscience 1990 38: 245 245

    Article  CAS  Google Scholar 

  20. Fearnley J.M., Lees A.J. . Ageing and Parkinson's disease: substantia nigra regional selectivity Brain 1991 114: 2283 2283

    Article  Google Scholar 

  21. Schulzer M. et al. A mathematical model of pathogenesis in idiopathic parkinsonism Brain 1994 117: 509 509

    Article  Google Scholar 

  22. Kozlowski D.A. et al. Delivery of a GDNF gene into the substantia nigra after a progressive 6-OHDA lesion maintains functional nigrostriatal connections Exp Neurol 2000 166: 1 1

    Article  CAS  Google Scholar 

  23. Connor B. et al. Differential effects of glial cell line-derived neurotrophic factor (GDNF) in the striatum and substantia nigra of the aged Parkinsonian rat Gene Therapy 1999 6: 1936 1936

    Article  CAS  Google Scholar 

  24. Winkler C., Sauer H., Lee C.S., Bjorklund A. . Short-term GDNF treatment provides long-term rescue of lesioned nigral dopaminergic neurons in a rat model of Parkinson's disease J Neurosci 1996 16: 7206 7206

    Article  CAS  Google Scholar 

  25. Natsume A. et al. Bcl-2 and GDNF delivered by HSV-mediated gene transfer act additively to protect dopaminergic neurons from 6-OHDA-induced degeneration Exp Neurol 2001 169: 231 231

    Article  CAS  Google Scholar 

  26. Rosenblad C., Kirik D., Bjorklund A. . Sequential administration of GDNF into the substantia nigra and striatum promotes dopamine neuron survival and axonal sprouting, but not striatal reinnervation or functional recovery in the partial 6-OHDA lesion model Exp Neurol 2000 161: 503 503

    Article  CAS  Google Scholar 

  27. McCown T.J. et al. Differential and persistent expression patterns of CNS gene transfer by an adeno-associated virus (AAV) vector Brain Res 1996 713: 99 99

    Article  CAS  Google Scholar 

  28. Kaplitt M.G. et al. Long-term gene expression and phenotypic correction using adeno-associated virus vectors in the mammalian brain Nat Genet 1994 8: 148 148

    Article  CAS  Google Scholar 

  29. Lo W.D. et al. Adeno-associated virus-mediated gene transfer to the brain: duration and modulation of expression Hum Gene Ther 1999 10: 201 201

    Article  CAS  Google Scholar 

  30. Ohta M. et al. Apomorphine up-regulates NGF and GDNF synthesis in cultured mouse astrocytes Biochem Biophys Res Commun 2000 272: 18 18

    Article  CAS  Google Scholar 

  31. Zhou J. et al. Differential expression of mRNAs of GDNF family in the striatum following 6-OHDA-induced lesion Neuroreport 2000 11: 3289 3289

    Article  CAS  Google Scholar 

  32. Chamberlin N.L., Du B., de Lacalle S., Saper C.B. . Recombinant adeno-associated virus vector: use for transgene expression and anterograde tract tracing in the CNS Brain Res 1998 793: 169 169

    Article  CAS  Google Scholar 

  33. Connor B. et al. Glial cell line-derived neurotrophic factor (GDNF) gene delivery protects dopaminergic terminals from degeneration Exp Neurol 2001 169: 83 83

    Article  CAS  Google Scholar 

  34. Kirik D., Georgievska B., Rosenblad C., Bjorklund A. . Delayed infusion of GDNF promotes recovery of motor function in the partial lesion model of Parkinson's disease Eur J Neurosci 2001 13: 1589 1589

    Article  CAS  Google Scholar 

  35. Akerud P. et al. Differential effects of glial cell line-derived neurotrophic factor and neurturin on developing and adult substantia nigra dopaminergic neurons J Neurochem 1999 73: 70 70

    Article  CAS  Google Scholar 

  36. Horger B.A. et al. Neurturin exerts potent actions on survival and function of midbrain dopaminergic neurons J Neurosci 1998 18: 4929 4929

    Article  CAS  Google Scholar 

  37. Rosenblad C. et al. In vivo protection of nigral dopamine neurons by lentiviral gene transfer of the novel GDNF-family member neublastin/artemin Mol Cell Neurosci 2000 15: 199 199

    Article  CAS  Google Scholar 

  38. Fan D. et al. Prevention of dopaminergic neuron death by adeno-associated virus vector-mediated GDNF gene transfer in rat mesencephalic cells in vitro Neurosci Lett 1998 248: 61 61

    Article  CAS  Google Scholar 

  39. Matsushita N. et al. Cloning and structural organization of the gene encoding the mouse glial cell line-derived neurotrophic factor, GDNF Gene 1997 203: 149 149

    Article  CAS  Google Scholar 

  40. Matsushita T. et al. Adeno-associated virus vectors can be efficiently produced without helper virus Gene Therapy 1998 5: 938 938

    Article  CAS  Google Scholar 

  41. Sawada H. et al. Neuroprotective mechanism of glial cell line-derived neurotrophic factor in mesencephalic neurons J Neurochem 2000 74: 1175 1175

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We thank Masashi Urabe and Dongsheng Fan for helpful advice. We also thank Avigen for providing the AAV vector production system. This study was supported in part by a Grant-in-Aid for Scientific Research on Priority Areas and Special Coordination Funds for Promoting Science and Technology from the Ministry of Education, Culture, Sports, Science and Technology, the Japanese Government; by Health Sciences Research Grants from the Ministry of Health Labour and Welfare of Japan; and by Core Research for Evolutional Science and Technology (CREST) of Japan Science and Technology Corporation (JST).

Author information

Authors and Affiliations

  1. Department of Neurology, Jichi Medical School, Japan

    L Wang, S Muramatsu, Y Lu, K Ikeguchi, K Fujimoto & I Nakano

  2. Division of Genetic Therapeutics, Center for Molecular Medicine, Jichi Medical School, Japan

    L Wang, Y Lu, T Okada, H Mizukami, Y Hanazono, A Kume & K Ozawa

  3. Institute for Comprehensive Medical Science, Fujita Health University, Japan

    F Urano, H Ichinose & T Nagatsu

Authors
  1. L Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. S Muramatsu
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Y Lu
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. K Ikeguchi
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. K Fujimoto
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. T Okada
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. H Mizukami
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Y Hanazono
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. A Kume
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. F Urano
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. H Ichinose
    View author publications

    You can also search for this author in PubMed Google Scholar

  12. T Nagatsu
    View author publications

    You can also search for this author in PubMed Google Scholar

  13. I Nakano
    View author publications

    You can also search for this author in PubMed Google Scholar

  14. K Ozawa
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Wang, L., Muramatsu, S., Lu, Y. et al. Delayed delivery of AAV-GDNF prevents nigral neurodegeneration and promotes functional recovery in a rat model of Parkinson's disease. Gene Ther 9, 381–389 (2002). https://doi.org/10.1038/sj.gt.3301682

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/sj.gt.3301682

Keywords

This article is cited by

Search

Advanced search

Quick links