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Functional changes in cocultures of mesencephalon and striatal neurons from embryonic C57/BL6 mice due to low concentrations of 1-Methyl-4-Phenylpyridinium (MPP+)

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Summary

1-Methyl-4-phenylpyridinium (MPP+), the active metabolite of 1-Methyl-4-phenyl-1, 2, 3, 6-tetrahydropyridine (MPTP) is taken up into dopaminergic terminals and selectively destroys dopaminergic neurons, serving as a valuable tool in animal model of Parkinson's disease. Cocultures from ventral mesencephalon and neostriatum of embryonic C57/BL6 mouse brains were used to study the sensitivity of dopaminergic neurons to the toxic agent MPP+. Cultures were grown for 9 days in vitro and exposed to different concentrations of MPP+ for various times. Treatment with (0.1–1.0 μM) MPP+ for 24 hours decreased3H-dopamine (3H-DA) uptake with an IC50 at 0.2 μM. Exposure of cells to 1 μM MPP+ over time decreased the+H-DA uptake to 38% of controls within the first two hours of incubation and to 8% after 48 hours. Loss of tyrosine hydroxylase (TH) positive cells became evident at 0.1 μM MPP+ (80% of control) leading to maximal toxicity at 10 μM (20% of control). MPP+ reduced the dopamine content in the cultures in a dose dependent manner (IC50 at 0.1 μM) and failed to show reversibility in recovery studies. These findings provide evidence that exposure of MPP+ even at low concentrations and for short time in our coculture model results in irreversible toxicity for dopaminergic neurons.

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References

  • Berger B, di Porzio U, Dagnet MC, Gay M, Vigny A, Glowinski J, Prochiantz A (1982) Long term development of mesencephalic dopaminergic neurons of mouse embryos in dissociated primary cultures: morphologic and histochemical characteristics. Neurosci Lett 7: 193–205

    Google Scholar 

  • Bernheimer H, Birkmayer W, Hornykiewicz O, Jellinger K, Seitelberger F (1973) Brain dopamine and the syndromes of Parkinson and Huntington. J Neurol Sci 20: 415–455

    PubMed  Google Scholar 

  • Burns RS, Chiueh CC, Markey SP, Ebert MH, Jacobowitz DM, Kopin IJ (1983) A primate model of parkinsonism: selective destruction of dopaminergic neuros in the pars compacta of the substantia nigra by N-methyl-4-phenyl-1,2,3,6-tetrahydropyridine. Proc Natl Acad Sci USA 80: 4546–4550

    PubMed  Google Scholar 

  • Coyle JT, Henry D (1973) Catecholamines in fetal and newborn rat brain. J Neurochem 21: 61–67

    PubMed  Google Scholar 

  • Danias P, Nicklas WJ, Ofori S, Shen J, Mytilineou C (1989) Mesencephalic dopamine neurons become less sensitive to 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine toxicity during development in vitro. J Neurochem 53: 1149–1155

    PubMed  Google Scholar 

  • Di Porzio U, Daguet M-C, Glowinski J, Prochiantz A (1980) Effect of striatal cells on in vitro maturation of mesencephalic dopaminergic neurons grown in serum-free conditions. Nature 288: 370–373

    PubMed  Google Scholar 

  • Engele J, Pilgrim C, Kirsch M, Reisert I (1989) Different developmental potentials of diencephalic and mesencephalic dopaminergic neurons in vitro. Brain Res 483: 98–109

    PubMed  Google Scholar 

  • Friedman L, Mytilineou C (1987) The toxicity of MPTP to dopamine neurons in culture is reduced at high concentrations. Neurosci Lett 79: 65–72

    PubMed  Google Scholar 

  • Hasegawa E, Takeshige K, Oishi T, Murai Y, Minakami S (1990) 1-Methyl-4-phenylpyr-idinium (MPP+) induces NADH-dependent Superoxide formation and enhances NADH-dependent lipid peroxidation in bovine heart submitochondrial particles. Biochem Biophys Res Commun 170(3): 1049–1055

    PubMed  Google Scholar 

  • Heikkila RE, Hess A, Durosin RC (1984) Dopaminergic neurotoxicity of 1,2,3,6-tetra-hydropyridine in mice. Science 224: 1451–1453

    PubMed  Google Scholar 

  • Javitch JA, D'Amato RJ, Stritmatter SM, Snyder SH (1985) Parkinsonism-inducing neurotoxin, N-methyl-4-phenyl-1,2,3,6-tetrahydropyridine: uptake of the metabolic N-methyl-4-phenylpyridine by dopamine neurons explains selective toxicity. Proc Natl Acad Sci USA 82: 2173–2177

    PubMed  Google Scholar 

  • Johannessen JN, Chiueh CC, Bacon JP, Garrick NA, Burns RS, Weise VK, Kopin I-J, Paresi JE, Marke P (1989) Effects of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine in the dog, effect of pargyline pretreatment. J Neurochem 53: 582–589

    PubMed  Google Scholar 

  • Langsten JW, Baliard P, Tetrud JW, Irwin I (1983) Chronic parkinsonism in humans due to a product of meperidine analogue synthesis. Science 219: 979–980

    PubMed  Google Scholar 

  • Levitt P, Pintar JE, Breakefield XO (1982) Immunocytochemical demostration of monoamine oxidase B in brain astrocytes and serotonergic neurons. Proc Natl Acad Sci USA 79: 6385–6389

    PubMed  Google Scholar 

  • Lindvall O, Björklund A (1983) Dopamine and norepinephrine containing neuron systems: their anatomy in the rat brain. In: Emson PC (ed) Chemical neuroanatomy. Raven Press, New York, pp 229–255

    Google Scholar 

  • Liu J, Lauder JM (1991) Serotonin and nialamide differentially regulate survival and growth of cultured serotonin and catecholamine neurons. Dev Brain Res 60: 59–67

    Google Scholar 

  • Michel PP, Dandapani BK, Knusel B, Sanchez-Ramos J, Hefti F (1990a) Toxicity of 1-methyl-4-phenylpyridinium for rat dopaminergic neurons in culture: selectivity and irreversability. J Neurochem 54: 1102–1109

    PubMed  Google Scholar 

  • Michel PP, Bhuvaneshari K, Dandapani BK, Efange MN, Hefti F (1990b) Potential environment neurotoxins related to 1-methyl-4-phenylpyridinium: selective toxicity of 1-methyl-4-phenylpyridinium and 1-methyl-4-cyclohexylpyridinium for dopaminergic neurons in culture. Exp Neurol 108: 141–150

    PubMed  Google Scholar 

  • Mytilineou C, Cohen G (1984) 1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridin destroys dopamine neurons in explants of rat embryo mesencephalon. Science 225: 224–226

    PubMed  Google Scholar 

  • Mytilineou C, Cohen G, Heikkila RE (1985) 1-Methyl-4-phenylpyridine (MPP+) is toxic to mesencephalic dopamine neurons in culture. Neurosci Lett 57: 19–24

    PubMed  Google Scholar 

  • Nicklas WJ, Vyas I, Heikkila RE (1985) Inhibition of NADH-linked oxidation in brain mitochondria by 1-methyl-4-phenylpyridine, a metabolite of the neurotoxin 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine. Life Sci 36: 2503–2508

    PubMed  Google Scholar 

  • Prochiantz A, di Porzio U, Kato A, Berger B, Glowinski J (1979) In vitro maturation of mesencephalic dopaminergic neurons from mouse embryos is enhanced in presence of their striatal target cells. Proc Natl Acad Sci USA 76: 5387–5391

    PubMed  Google Scholar 

  • Ramsay RR, Singer TP (1986) Energy-dependent uptake of N-Methyl-4-phenylpyridinium, the neurotoxic metabolite of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine, by mitochondria. J Biol Chem 261(17): 7585–7587

    PubMed  Google Scholar 

  • Sanchez-Ramos JR, Barrett JN, Goldstein M, Weiner WJ, Hefti F (1987) 1-Methyl-4-phenylpyridinium (MPP+) but not 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) selectively destroys dopaminergic neurons in culture of dissociated rat mesencephalic neurons. Neurosci Lett 72: 215–220

    Google Scholar 

  • Sanchez-Ramos JR, Michel P, Weiner WJ, Hefti F (1988) Selective destruction of cultured dopaminergic neurons from fetal rat mesencephalon by 1-methyl-4-phenylpyridinium: cytochemical and morphological evidence. J Neurochem 50: 1934–1944

    PubMed  Google Scholar 

  • Sanchez-Ramos JR, Song S, Mash DC, Weiner WJ (1992) 21-Aminosteroids interact with the dopamine transporter to protect against 1-methyl-4-phenylpyridinium-induced neurotoxicity. J Neurochem 58: 328–334

    PubMed  Google Scholar 

  • Schambra UB, Lauder JM, Silver J (1992) Atlas of the prenatal mouse brain. Academic Press, London, pp 76–84, 108–112

    Google Scholar 

  • Schinelli S, Zuddas A, Kopin IJ, Barker JL, di Porzio U (1988) 1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine metabolism and 1-methyl-4-phenylpyridinium uptake in dissociated cell cultures from the embryonic mesencephalon. J Neurochem 50: 1900–1907

    PubMed  Google Scholar 

  • Schneider JS, Markam CH (1986) Neurotoxic effects of N-methyl-4-phenyl-1,2,3,6-tetra-hydropyridine (MPTP) in the cat. Tyrosine hydroxylase immunohistochemistry. Brain Res 373: 258–267

    PubMed  Google Scholar 

  • Sebben M, Gabrion J, Manzoni O, Sladeczek F, Gril C, Bockaert J, Dumuis A (1990) Establishment of a long-term primary culture of striatal neurons. Dev Brain Res 52: 229–239

    Google Scholar 

  • Sofie E (1986) Untersuchung von biogenen Aminen, Metaboliten, Ascorbinsäure und Glutathion mittels HPLC-ECD und deren Verhalten in ausgewählten Lebensmitteln und im Organismus von Tier und Mensch. Dissertation, Technische Universität Wien

    Google Scholar 

  • Sumners C, Phillips MI, Raizada MK (1983) Rat brain cells in primary culture: visualization and measurement of catecholamines. Brain Res 264: 267–275

    PubMed  Google Scholar 

  • Tomozawa Y, Appel SH (1986) Soluble striatal extracts development of mesencephalic dopaminergic neurons in vitro. Brain Res 399: 111–124

    PubMed  Google Scholar 

  • Turrens JF, Boveris A (1980) Generation of superoxide anion by the NADH-dehydrogenase of bovine heart mitochondria. Biochem J 191: 421–427

    PubMed  Google Scholar 

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  1. E. Koutsilieri

    Present address: Institute of Medical Chemistry, Veterinary University of Vienna, Austria

Authors and Affiliations

  1. Institute of Medical Chemistry, Veterinary University of Vienna, Austria

    W. W. Chan, D. Reinitzer & W. -D. Rausch

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  1. E. Koutsilieri
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  2. W. W. Chan
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  3. D. Reinitzer
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  4. W. -D. Rausch
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Koutsilieri, E., Chan, W.W., Reinitzer, D. et al. Functional changes in cocultures of mesencephalon and striatal neurons from embryonic C57/BL6 mice due to low concentrations of 1-Methyl-4-Phenylpyridinium (MPP+). J. Neural Transmission 94, 189–197 (1993). https://doi.org/10.1007/BF01277024

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