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Vol. 1, No. 4-5, 2004
Issue release date: November 2004
Section title: Paper
Neurodegenerative Dis 2004;1:175–183
(DOI:10.1159/000080983)

Caenorhabditis elegans MPP+ Model of Parkinson’s Disease for High-Throughput Drug Screenings

Braungart E.a · Gerlach M.b · Riederer P.b · Baumeister R.c · Hoener M.C.d
aPieris Proteolab AG, Freising-Weihenstephan, bDepartment of Psychiatry, University of Würzburg, Würzburg, and cBio3/Bioinformatics and Molecular Genetics, University of Freiburg, Freiburg, Germany; dF. Hoffmann-La Roche, Basel, Switzerland
email Corresponding Author

Abstract

The neurotoxin MPTP and its active metabolite MPP+ cause Parkinson’s disease (PD)-like symptoms in vertebrates by selectively destroying dopaminergic neurons in the substantia nigra. MPTP/MPP+ models have been established in rodents to screen for pharmacologically active compounds. In addition to being costly and time consuming, these animal models are not suitable for large scale testings using compound libraries. We present a novel MPP+-based model for high-throughput screenings using the nematode Caenorhabditis elegans. Incubation of C. elegans with MPTP or its active metabolite MPP+ resulted in strong symptomatic defects including reduced mobility and increased lethality, and is correlated with a specific degeneration of the dopaminergic neurons. The phenotypic consequences of MPTP/MPP+ treatments were recorded using automated hardware and software for quantification. Incubation of C. elegans with a variety of pharmacologically active components used in PD treatment reduced the MPP+-induced defects. Our data suggest that the C. elegans MPTP/MPP+ model can be used for the quantitative evaluation of anti-PD drugs.

© 2004 S. Karger AG, Basel


  

Key Words

  • Caenorhabditis elegans
  • Dopaminergic neurons
  • High-throughput screenings
  • MPP+
  • Parkinson’s disease
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References

  1. Nass R, Hall DH, Miller DM 3rd, Blakely RD: Neurotoxin-induced degeneration of dopamine neurons in Caenorhabditis elegans. Proc Natl Acad Sci USA 2002;99:3264–3269.
  2. Gerlach M, Riederer P: Animal models of Parkinson’s disease: An empirical comparison with the phenomenology of the disease in man. J Neural Transm 1996;103:987–1041.
  3. Kindt MV, Heikkila RE: Prevention of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine-induced dopaminergic toxicity in mice by MDL 72145, a selective inhibitor of MAO-B. Life Sci 1986;38:1459–1462.
  4. Russ H, Staust K, Martel F, Gliese M, Schomig E: The extraneuronal transporter for monoamine transmitters exists in cells derived from human central nervous system glia. Eur J Neurosci 1996;8:1256–1264.
  5. Javitch JA, D’Amato RJ, Strittmatter SM, Snyder SH: Parkinsonism-inducing neurotoxin, N-methyl-4-phenyl-1,2,3,6-tetrahydropyridine: Uptake of the metabolite N-methyl-4- phenylpyridine by dopamine neurons explains selective toxicity. Proc Natl Acad Sci USA 1985;82:2173–2177.
  6. Mizuno Y, Suzuki K, Sone N, Saitoh T: Inhibition of mitochondrial respiration by 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) in mouse brain in vivo. Neurosci Lett 1988;91:349–353.
  7. Tipton KF, Singer TP: Advances in our understanding of the mechanisms of the neurotoxicity of MPTP and related compounds. J Neurochem 1993;61:1191–1206.
  8. Smith MT, Ekstrom G, Sandy MS, Di Monte D: Studies on the mechanism of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine cytotoxicity in isolated hepatocytes. Life Sci 1987;40:741–748.
  9. Burns RS, Chiueh CC, Markey SP, Ebert MH, Jacobowitz DM, Kopin IJ: A primate model of parkinsonism: Selective destruction of dopaminergic neurons in the pars compacta of the substantia nigra by N-methyl-4-phenyl-1,2,3,6-tetrahydropyridine. Proc Natl Acad Sci USA 1983;80:4546–4550.
  10. Herrero MT, Augood SJ, Asensi H, Hirsch EC, Agid Y, Obeso JA, Emson PC: Effects of L-DOPA-therapy on dopamine D2 receptor mRNA expression in the striatum of MPTP-intoxicated parkinsonian monkeys. Brain Res Mol Brain Res 1996;42:149–155.
  11. Taylor JR, Elsworth JD, Roth RH, Sladek JR Jr, Redmond DE Jr: Severe long-term 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine-induced parkinsonism in the vervet monkey (Cercopithecus aethiops sabaeus). Neuroscience 1997;81:745–755.
  12. Doudet DJ, Chan GL, Holden JE, McGeer EG, Aigner TA, Wyatt RJ, Ruth TJ: 6-[18F]Fluoro-L-DOPA PET studies of the turnover of dopamine in MPTP-induced parkinsonism in monkeys. Synapse 1998;29:225–232.
  13. Heikkila RE: Differential neurotoxicity of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) in Swiss-Webster mice from different sources. Eur J Pharmacol 1985;117:131–133.
  14. Jarvis MF, Wagner GC: Neurochemical and functional consequences following 1-methyl- 4-phenyl-1,2,5,6-tetrahydropyridine (MPTP) and methamphetamine. Life Sci 1985;36:249–254.
  15. Bradbury AJ, Costall B, Domeney AM, Jenner P, Kelly ME, Marsden CD, Naylor RJ: 1-Methyl-4-phenylpyridine is neurotoxic to the nigrostriatal dopamine pathway. Nature 1986;319:56–57.
  16. Frohna PA, Rothblat DS, Joyce JN, Schneider JS: Alterations in dopamine uptake sites and D1 and D2 receptors in cats symptomatic for and recovered from experimental parkinsonism. Synapse 1995;19:46–55.
  17. Bargmann CI: Neurobiology of the Caenorhabditis elegans genome. Science 1998;282:2028–2033.
  18. Baumeister R, Ge L: The worm in us – Caenorhabditis elegans as a model of human disease. Trends Biotechnol 2002;20:147–148.
  19. White JG, Southgate E, Thomson JN, Brenner S: The structure of the nervous system of the nematode Caenorhabditis elegans. Philos Trans R Soc Lond 1986;314:1–340.
  20. Sulston J, Dew M, Brenner S: Dopaminergic neurons in the nematode Caenorhabditis elegans. J Comp Neurol 1975;163:215–226.
  21. Wintle RF, Van Tol HH: Dopamine signaling in Caenorhabditis elegans-potential for parkinsonism research. Parkinsonism Relat Disord 2001;7:177–183.
  22. Wood WB: The Nematode C. elegans. Cold Spring Harbor, Cold Spring Harbor Laboratory, 1988.
  23. Lints R, Emmons SW: Patterning of dopaminergic neurotransmitter identity among Caenorhabditis elegans ray sensory neurons by a TGFbeta family signaling pathway and a Hox gene. Development 1999;126:5819–5831.
  24. Hobert O, Mori I, Yamashita Y, Honda H, Ohshima Y, Liu Y, Ruvkun G: Regulation of interneuron function in the C. elegans thermoregulatory pathway by the ttx-3 LIM homeobox gene. Neuron 1997;19:345–357.
  25. Troemel ER, Kimmel BE, Bargmann CI: Reprogramming chemotaxis responses: Sensory neurons define olfactory preferences in C. elegans. Cell 1997;91:161–169.
  26. Rohrig S, Rockelein I, Donhauser R, Baumeister R: Protein interaction surface of the POU transcription factor UNC-86 selectively used in touch neurons. EMBO J 2000;19:3694–3703.
  27. Song X, Perkins S, Jortner BS, Ehrich M: Cytotoxic effects of MPTP on SH-SY5Y human neuroblastoma cells. Neurotoxicology 1997;18:341–353.
  28. Marini AM, Schwartz JP, Kopin IJ: The neurotoxicity of 1-methyl-4-phenylpyridinium in cultured cerebellar granule cells. J Neurosci 1989;9:3665–3672.
  29. Sanchez-Ramos JR, Song S, Facca A, Basit A, Epstein CJ: Transgenic murine dopaminergic neurons expressing human Cu/Zn superoxide dismutase exhibit increased density in culture, but no resistance to methylphenylpyridinium-induced degeneration. J Neurochem 1997;68:58–67.
  30. Sawin ER, Ranganathan R, Horvitz HR: C. elegans locomotory rate is modulated by the environment through a dopaminergic pathway and by experience through a serotonergic pathway. Neuron 2000;26:619–631.
  31. Jenner P, Marsden CD: The actions of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine in animals as a model of Parkinson’s disease. J Neural Transm Suppl 1986;20:11–39.
  32. Olanow CW: The role of dopamine agonists in the treatment of early Parkinson’s disease. Neurology 2002;58:S33–41.
  33. Schafer WR, Kenyon CJ: A calcium-channel homologue required for adaptation to dopamine and serotonin in Caenorhabditis elegans. Nature 1995;375:73–78.
  34. Kamath RS, Fraser AG: Systematic functional analysis of the Caenorhabditis elegans genome using RNAi. Nature 2003;421:231–237.

  

Author Contacts

Ralf Baumeister, PhD
Bio3/ Bioinformatics and Molecular Genetics
Schaenzlestrasse 1
DE–79104 Freiburg (Germany)
Tel. +49 761 203 2799, Fax +49 761 203 8351, E-Mail baumeister@celegans.de

  

Article Information

Number of Print Pages : 9
Number of Figures : 5, Number of Tables : 1, Number of References : 34

  

Publication Details

Neurodegenerative Diseases

Vol. 1, No. 4-5, Year 2004 (Cover Date: Released November 2004)

Journal Editor: R.M. Nitsch, Zürich; C. Hock, Zürich
ISSN: 1660–2854 (print), 1660–2862 (Online)

For additional information: http://www.karger.com/ndd


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Drug Dosage: The authors and the publisher have exerted every effort to ensure that drug selection and dosage set forth in this text are in accord with current recommendations and practice at the time of publication. However, in view of ongoing research, changes in goverment regulations, and the constant flow of information relating to drug therapy and drug reactions, the reader is urged to check the package insert for each drug for any changes in indications and dosage and for added warnings and precautions. This is particularly important when the recommended agent is a new and/or infrequently employed drug.
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References

  1. Nass R, Hall DH, Miller DM 3rd, Blakely RD: Neurotoxin-induced degeneration of dopamine neurons in Caenorhabditis elegans. Proc Natl Acad Sci USA 2002;99:3264–3269.
  2. Gerlach M, Riederer P: Animal models of Parkinson’s disease: An empirical comparison with the phenomenology of the disease in man. J Neural Transm 1996;103:987–1041.
  3. Kindt MV, Heikkila RE: Prevention of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine-induced dopaminergic toxicity in mice by MDL 72145, a selective inhibitor of MAO-B. Life Sci 1986;38:1459–1462.
  4. Russ H, Staust K, Martel F, Gliese M, Schomig E: The extraneuronal transporter for monoamine transmitters exists in cells derived from human central nervous system glia. Eur J Neurosci 1996;8:1256–1264.
  5. Javitch JA, D’Amato RJ, Strittmatter SM, Snyder SH: Parkinsonism-inducing neurotoxin, N-methyl-4-phenyl-1,2,3,6-tetrahydropyridine: Uptake of the metabolite N-methyl-4- phenylpyridine by dopamine neurons explains selective toxicity. Proc Natl Acad Sci USA 1985;82:2173–2177.
  6. Mizuno Y, Suzuki K, Sone N, Saitoh T: Inhibition of mitochondrial respiration by 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) in mouse brain in vivo. Neurosci Lett 1988;91:349–353.
  7. Tipton KF, Singer TP: Advances in our understanding of the mechanisms of the neurotoxicity of MPTP and related compounds. J Neurochem 1993;61:1191–1206.
  8. Smith MT, Ekstrom G, Sandy MS, Di Monte D: Studies on the mechanism of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine cytotoxicity in isolated hepatocytes. Life Sci 1987;40:741–748.
  9. Burns RS, Chiueh CC, Markey SP, Ebert MH, Jacobowitz DM, Kopin IJ: A primate model of parkinsonism: Selective destruction of dopaminergic neurons in the pars compacta of the substantia nigra by N-methyl-4-phenyl-1,2,3,6-tetrahydropyridine. Proc Natl Acad Sci USA 1983;80:4546–4550.
  10. Herrero MT, Augood SJ, Asensi H, Hirsch EC, Agid Y, Obeso JA, Emson PC: Effects of L-DOPA-therapy on dopamine D2 receptor mRNA expression in the striatum of MPTP-intoxicated parkinsonian monkeys. Brain Res Mol Brain Res 1996;42:149–155.
  11. Taylor JR, Elsworth JD, Roth RH, Sladek JR Jr, Redmond DE Jr: Severe long-term 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine-induced parkinsonism in the vervet monkey (Cercopithecus aethiops sabaeus). Neuroscience 1997;81:745–755.
  12. Doudet DJ, Chan GL, Holden JE, McGeer EG, Aigner TA, Wyatt RJ, Ruth TJ: 6-[18F]Fluoro-L-DOPA PET studies of the turnover of dopamine in MPTP-induced parkinsonism in monkeys. Synapse 1998;29:225–232.
  13. Heikkila RE: Differential neurotoxicity of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) in Swiss-Webster mice from different sources. Eur J Pharmacol 1985;117:131–133.
  14. Jarvis MF, Wagner GC: Neurochemical and functional consequences following 1-methyl- 4-phenyl-1,2,5,6-tetrahydropyridine (MPTP) and methamphetamine. Life Sci 1985;36:249–254.
  15. Bradbury AJ, Costall B, Domeney AM, Jenner P, Kelly ME, Marsden CD, Naylor RJ: 1-Methyl-4-phenylpyridine is neurotoxic to the nigrostriatal dopamine pathway. Nature 1986;319:56–57.
  16. Frohna PA, Rothblat DS, Joyce JN, Schneider JS: Alterations in dopamine uptake sites and D1 and D2 receptors in cats symptomatic for and recovered from experimental parkinsonism. Synapse 1995;19:46–55.
  17. Bargmann CI: Neurobiology of the Caenorhabditis elegans genome. Science 1998;282:2028–2033.
  18. Baumeister R, Ge L: The worm in us – Caenorhabditis elegans as a model of human disease. Trends Biotechnol 2002;20:147–148.
  19. White JG, Southgate E, Thomson JN, Brenner S: The structure of the nervous system of the nematode Caenorhabditis elegans. Philos Trans R Soc Lond 1986;314:1–340.
  20. Sulston J, Dew M, Brenner S: Dopaminergic neurons in the nematode Caenorhabditis elegans. J Comp Neurol 1975;163:215–226.
  21. Wintle RF, Van Tol HH: Dopamine signaling in Caenorhabditis elegans-potential for parkinsonism research. Parkinsonism Relat Disord 2001;7:177–183.
  22. Wood WB: The Nematode C. elegans. Cold Spring Harbor, Cold Spring Harbor Laboratory, 1988.
  23. Lints R, Emmons SW: Patterning of dopaminergic neurotransmitter identity among Caenorhabditis elegans ray sensory neurons by a TGFbeta family signaling pathway and a Hox gene. Development 1999;126:5819–5831.
  24. Hobert O, Mori I, Yamashita Y, Honda H, Ohshima Y, Liu Y, Ruvkun G: Regulation of interneuron function in the C. elegans thermoregulatory pathway by the ttx-3 LIM homeobox gene. Neuron 1997;19:345–357.
  25. Troemel ER, Kimmel BE, Bargmann CI: Reprogramming chemotaxis responses: Sensory neurons define olfactory preferences in C. elegans. Cell 1997;91:161–169.
  26. Rohrig S, Rockelein I, Donhauser R, Baumeister R: Protein interaction surface of the POU transcription factor UNC-86 selectively used in touch neurons. EMBO J 2000;19:3694–3703.
  27. Song X, Perkins S, Jortner BS, Ehrich M: Cytotoxic effects of MPTP on SH-SY5Y human neuroblastoma cells. Neurotoxicology 1997;18:341–353.
  28. Marini AM, Schwartz JP, Kopin IJ: The neurotoxicity of 1-methyl-4-phenylpyridinium in cultured cerebellar granule cells. J Neurosci 1989;9:3665–3672.
  29. Sanchez-Ramos JR, Song S, Facca A, Basit A, Epstein CJ: Transgenic murine dopaminergic neurons expressing human Cu/Zn superoxide dismutase exhibit increased density in culture, but no resistance to methylphenylpyridinium-induced degeneration. J Neurochem 1997;68:58–67.
  30. Sawin ER, Ranganathan R, Horvitz HR: C. elegans locomotory rate is modulated by the environment through a dopaminergic pathway and by experience through a serotonergic pathway. Neuron 2000;26:619–631.
  31. Jenner P, Marsden CD: The actions of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine in animals as a model of Parkinson’s disease. J Neural Transm Suppl 1986;20:11–39.
  32. Olanow CW: The role of dopamine agonists in the treatment of early Parkinson’s disease. Neurology 2002;58:S33–41.
  33. Schafer WR, Kenyon CJ: A calcium-channel homologue required for adaptation to dopamine and serotonin in Caenorhabditis elegans. Nature 1995;375:73–78.
  34. Kamath RS, Fraser AG: Systematic functional analysis of the Caenorhabditis elegans genome using RNAi. Nature 2003;421:231–237.