MPTP-Induced Modulation of Neurotransmitters in SH-SY5Y Human Neuroblastoma Cells

Abstract

Neurotoxic effects of MPTP(1-methyl-4-phenyl-1, 2, 3, 6-tetrahydropyridine) were evaluated in vitro using a human neuronal cell line, SH-SY5Y, that contained features contributing to expression of MPTP toxicity in vivo, namely, a transport system for dopam ine (DA) and monam ine oxidase (MAO) activity. In this model system, MPTP was found to reduce levels of catecholamines (DA, norepinephrine, epinephrine), serotonin (5-HT), and the 5-HT metabolite 5-hydroxyindoleacetic acid (5-HIAA). MPTP enhanced 3H-DA release, which could contribute to the reduction in DA concentrations seen in these cells. In addition, MPTP inhibited MAO activity (K_i 2.26 X 10-5 M). Pretreatment with the MAO inhibitor pargy-line protected the cells from MPTP-induced alterations of catecholamines and the decrease in 5-HT. In this in vitro model, the cholinergic antagonists atro-pine and A-tubocurarine also protected cells from MPTP-induced alterations of catecholamines. The capability of cholinergic antagonists to prevent the MPTP-induced alterations of catecholamine concentrations suggests a possible cholinergic contribution to MPTP neurotoxicity in this cell line.

Keywords

in vitro studies MPP+MPTP SH-SY5Yhuman neuroblastoma cells

References

Adem, A. , M. E. K. Mattsson , A. Nordberg , and S. Pahlman . 1987. Muscarinic receptors in human SH-SY5 Yneuroblastoma cell line: regulation by phorbol ester and retinoic acid-induced differentiation. Dev. Brain Res. 33:235–242.

Biedler, J. L. , L. Halson and B. A. Spengler . 1973. Morphology and growth, tumorigenicity, and cytogenecities of human neuroblastoma cells in continuous culture. Cancer Res. 33:2643–2652.

Biedler, J. L. , S. Roffler-Tarlov , M. Schachner , and L. S. Freedman . 1978. Multiple neurotransmitter synthesis by human neuroblastoma cell lines and clones. Cancer Res. 38:3751–3757.

Bocchetta, A. , M. P. Piccardi , M. del Zompo , S. Pintus , and G. U. Corsini . 1985. 1-Methyl-4-phenyl-1, 2, 3, 6-tetrahydropyridine: Correspondence of its binding sites to monoamine oxidase in rat brain, and inhibition of dapomine oxidative deamination in vivo and in vitro. J. Neurochem. 45:673–676.

Burns, R. S. , C. C. Chiueh , S. P. Markey , M. H. Ebert , D. M. Jacobowitz and I. J. Kopin . 1983. A primate model of parkinsonsonism: 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 80:4546–4550.

Chiba, K. , L. A. Peterson , K. P. Castagnoli , A. J. Trevor , and N. E. Castagnoli, Jr . 1985. Studies on the molecular mechanism of bioactivation of the selective nigrostriatal toxin l-methyl-4-phenyl-1, 2, 3, 6-tetrahydropyridine. Drug Metab. Disposit. 13:342–347.

Chiba, K. , A. Trevor , and N. Castagnoli . 1984. Metabolism of the neurotoxic amine, MPTP, by brain monoamine oxidase. Biochem. Biophys. Res. Com-mun. 120:574–579.

Chiueh, C. C. , R. S. Burns , S. P. Markey , D. M. Jacobowitz and I. J. Kopin . 1985. Primate model of parkinsonism: Selective lesion of nigrostriatal neurons by 1-methyl-4-phenyl-1, 2, 3, 6-tetrahydropyridine produces an extrapyramidal syndrome in rhesus monkeys. Life Sci. 36:213–218.

Chiueh, C. C. , S. P. Markey , R. E. Burns , J. N. Johannessen , A. Pert and I. J. Kopin . 1984. Neurochemical and behavioral effects of systemic and intran-igral administration of 1-methyl-4-phenyl-1, 2, 3, 6-tetrahydropyridine in the rat. Eur. J. Pharmacol. 100:189–194.

10.

Cohen, G. , P. Paski , B. Cohen , A. Leist , C. Mystilineou , and M. D. Yahr . 1984. Pargylineand deprenyl prevent the neurotoxicity of 1-methyl-4-phenyl-1, 2, 3, 6-tetrahydropyridine (MPTP) in monkey. Eur. J. Pharmacol. 106:209–210.

11.

Davis, G. C. , A. C. Williams , S. P. Markey , M. H. Ebert , E. D. Caine , C. M. Beichert and I. J. Kopin . 1979. Chronic parkinsonism secondary to intravenous injection of meperidine analogues. Psychiatry Res. 1:249–254.

12.

Desole, M. S. , G. Esposito , P. Enrico , M. Miele , M. Fresu , G. de Natale , E. Miele , and G. Grella . 1993. Effects of aging on 1-methyl-4-phenyl-1, 2, 3, 6-tetrahydropyridine (MPTP) neurotoxic effects on striatum and brainstem in the rat. Neurosci. Lett. 159:143–146.

13.

DiMonte, D. A. , E. Y. Wu , L. Irwin , L. E. Delanney and J. W. Langston . 1992. Production and disposition of 1 -methyl-4-phenylpyridine in primary cultures of mouse astrocytes. Glia 5:48–55.

14.

Drug facts and comparisons. 1998. Antiparkinson agents, 289b-290n. St. Louis, MO: Facts and Comparisons.

15.

Elsworth, J. D. , A. Y. Deutch , D. E. Redmond , J. R. Sladek and R. H. Roth . 1987. Differential responsiveness to 1-methyl-4-phenyl-1, 2, 3, 6-tetrahydropyridine toxicity in subregions of the primate substania nigra and striatum. Life Sci. 40:193–202.

16.

Forno, L. S. 1990. Pathology of Parkinson's disease: The importance of the sub-stantia nigra and Lewy bodies. In Parkinson's Disease, ed. G. M. Stern , 185–238, Baltimore, MD: Johns Hopkins University Press.

17.

Frazer, A. , and J. G. Hensler . 1995. Serotonin. In Basic neurochemistry, ed. J. Siegel , B. W. Agranoff , R. W. Albert , and P. B. Molinoff , 283–308, New York, NY: Raven Press.

18.

Gaspar, P. , A. Febvret , and J. Colombo . 1993. Serotogenic sprouting in primate MPTP-induced hemiparkinsonism. Exp. Brain Res. 96:100–106.

19.

Giorguieff, M. F. , M. L. Lefloch , J. Glowinski and M. B. Beeson . 1977. Involvement of cholinergic presynaptic receptors of nicotinic and muscarinic types in the control of spontaneous release of dopamine from striatal dopaminergic terminals in the rat. J. Pharmacol. Exp. Ther. 200:535–544.

20.

Glover, V. , C. Gibb , and M. Sandier . 1986. Monoamine oxidase B (MAO-B) is the major catalyst for 1-methyl-4-phenyl-1, 2, 3, 6-tetrahydropyridine (MPTP) oxidation in human brain and other tissues. Neurosci. Lett. 64:216–220.

21.

Gould, J. , H. Reeve , P. F. T. Vaughan , and C. Peers . 1992. Nicotinic acetylcholine receptors in human neuroblastoma (SH-SY5Y) cells. Neurosci. Lett. 145:201–204.

22.

Hadjiconstantinou, M. , D. Cavalla , E. Anthoupoulou , H. E. Laird , and N. H. Neff . 1985. N-Methyl-4-phenyl-1, 2, 3, 6-tetrahydropyridine increases acetylcholine and decreases dopamine in mouse striatum: Both responses are blocked by anticholinergic drugs. J. Neurochem. 45:195–197.

23.

Heikkila, R. E. , L. Manzino , F. E. Cabbat and R. C. Duvoisin . 1984. Protection against the dopaminergic neurotoxicity of 1-methyl-4-phenyl-1, 2, 3, 6-tetrahydrophyridine by monoamine oxidase inhibitors. Nature 311:467–469.

24.

Hoppel, C. L. , D. Grinblattm , H. C. Kwokm , P. K. Arora , M. P. Singh and L. M. Sayre . 1987. Inhibition of mitochondrial respiration by analogs of 4-phenyl-pyridine and 1-methyl-4-phenylpyridinium (MPP+), theneurotoxic metabolite of MPTP. Biochem. Biophys. Res. Commun. 148:684–690.

25.

Itzhak, Y. , D. Mash , S.Zhang , and I. Stein . 1991. Characterization of N-methyl-4-phenyl-1, 2, 3, 6-tetrahydropyridine (MPTP) binding sites in C57BL/6 mouse brain: Mutual effects of monoamine oxidase inhibitors and δligands on MPTP and δbinding sites. Molec. Pharmacol. 39:385–393.

26.

Javitch, J. A. , R. J.D'Amato , S.M. Strittmatter , and S.H. Snyder . 1985. Parkin-sonian-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 82:2173–2177.

27.

Kinemuchi, H. , Y. Arai , and Y. Toyoshima . 1985. Participation of brain monoamine oxidase B form in the neurotoxicity of 1 -methyl-4-phenyl-1, 2, 3, 6-tetra-hydropyridine: Relationship between the enzyme inhibition and the neurotoxicity. Neurosci. Lett. 58:195–200.

28.

Koman, A. , S. Cazaubon , A. Adem , P. O. Courand , and A. D. Strosberg . 1993. Different regulatory patterns of M1 and M2 muscarinic receptor subtype RNA in SH-SY5Y human neuroblastoma induced by phorbol ester or DMSO. Neurosci. Lett. 149:79–82.

29.

Krueger, M. J. , K. McKeown , R. R. Ramsay , S. K. Youngster and T. P. Singer . 1990. Mechanism-based inactivation of monoamine oxidases A and B by tetrahydropyridines and dihydropyridines. Biochem. J. 268:219–224.

30.

Kukkonen, J. , P. Ojala , J. Nasman , H. Hamalainen , J. Heikkila , and K. E. O. Akerman . 1992. Muscarinic receptor subtypes in human neuroblastoma cell lines SH-SY5Y and IMR-32 as determined by receptor binding, Ca++ mobilization and northern blotting. J. Pharmacol. Exp. Ther. 263:1487–1493.

31.

Lai, M. , H. Griffiths , H. Pall , A. Willimas , and J. Lunec . 1993. An investigation into the role of reactive oxygen species in the mechanism of 1-methyl-4-phenyl-1, 2, 3, 6-tetrahydropyridine toxicity using neuronal cell lines. Biochem. Pharmacol. 45:927–933.

32.

Langston, J. W. , P. Ballard , J. W. Tetrud and I. Irwin . 1983. Chronic parkin-sonism in human due to a product of meperidine-analog synthesis. Science 219:979–980.

33.

Langston, J. W. , L. S. Forno , C. S. Rebert and I. Irwin . 1984a. Selective nigra toxicity after systemic administration of 1-methyl-4-phenyl-1, 2, 3, 6-tetra-hydropyridine (MPTP) in the squirrel monkey. Brain Res. 292:390–394.

34.

Langston, J. W. , I. Irwin , E. B. Langston and L. S. Forno . 1984b. 1-Methyl-4-phenylpyridinium ion (MPP+): Identification of a metabolite of MPTP, a toxin selective to the substantia nigra. Neurosci. Lett. 48:87–92.

35.

Langston, J. W. , I. Irwin , E. B. Langston and L. S. Forno . 1984c. Pargyline prevents MPTP-induced parkinsonism in primates. Science 225:1480–1482.

36.

Lukas, R. J. , S. A. Norman and L. Lucero . 1993. Characterization of nicotine acetylcholine receptors expressed by cells of the SH-SY5Y human neuroblastoma clonal line. Molec. Cell Neurosci. 4:1–12.

37.

Marini, A. M. , J. P. Schwartz and I. J. Kopin . 1989. The neurotoxicity of 1-methyl-4-phenylpyridinium in cultured cerebellar granule cells. J. Neurosci. 9:3665–3672.

38.

May, T. 1993. 1-Methyl-4-phenylpyridinium (MPP+) binds with high affinity to a β-carboline binding site located on monoamine oxidase type A in rat brain. Neurosci. Lett. 162:55–58.

39.

Mogi, M. , M. Harada , K. Kiuchi , and T. Nagatsu . 1988. Effects of administration of 1-methyl-4-phenyl-1, 2, 3, 6-tetrahydropyridine to mice on tyrosine hydroxylase, L-3,4-dihydroxyphenylamine decarboxylase, dopamine b-hydro-xylase, and monoamine oxidase activity in the striatum and hypothalamus. J. Neurochem. 50:1053–1056.

40.

Murphy, N. P. , S. G. Ball and P. F. T. Vaughan . 1991. Potassium- and carbachol-evoked release of [3H]noradrenaline from human neuroblastoma cells, SH-SY5Y J. Neurochem. 56:1810–1815.

41.

Mytilineuou, C. , G. Cohen , and R. Heikkila . 1985. 1-Methyl-4-phenylpyridine (MMP+) is toxic to mesencephalic dopamine neurons in culture. Neurosci. Lett. 57:19–24.

42.

Nagatsu, T. , T. Nakano , and H. Higashida . 1981. Expression of A and B types of monoamine oxidase in neuroblastoma hybrid cells. Neurochem. Int. 3:137–142.

43.

Naoi, M. , H. Ichinose , T. Takahashi , I. Nakatsu , and T. Nagatsu . 1988. Reduction of aromatic L-amino acid decarboxylase activity in clonal pheochromocy-toma PC12 cells by culture in the presence of N-methyl-4-phenylpyridinium ion (MPP+). Neurosci. Lett. 95:229–235.

44.

Naoi, M. , H. Suzuki , K. Kiuchi , T. Takahashi , and T. Nagatsu . 1987a. Effects of N-methyl-4-phenylpyridium ion on monoamine oxidase in a clonal rat pheochromocytoma cell line, PC12h. J. Neurochem. 48:1912–1916.

45.

Naoi, M. N. , T. Takahashi and T. Nagatsu . 1987b. Metabolism of N-methyl-4-phenyl-1, 2, 3, 6-tetrahydropyridinine in a clonal rat pheochromocytoma cell line, PC 12h. Life Sci. 41:2655–2661.

46.

Nicklas, W. J. , I. Vyas and R. E. Heikkila . 1985. Inhibition of NADH-linked oxidation in brain mitochondria by 1-methyl-4-phenylpyridium, a metabolite of the neurotoxin 1-methyl-4-phenyl-1, 2, 3, 6-tetrahydropyridine. Life Sci. 36:2503–2508.

47.

Nwanze, E. , F. Souverbie , G. Jos son , and E. Sundstrom . 1995. Regional biotrans-formation of MPTP in the CNS of rodents and its relation to neurotoxicity. NeuroToxicology 16:469–478.

48.

Ogata, T. , C. Myers , and J. L. Blumer . 1992. Determination of platelet monoamine oxidase activity by high-performance liquid chromatography with electrochemical detection. J. Chromatography 575:39–49.

49.

Pearce, J. M. S. 1992. Neurotransmitters and diagnostic markers. In Parkinson's Disease and its management, ed. J. M. S. Pearce , 53–62. Oxford, UK: Oxford University Press.

50.

Peterson, L. A. , P. S. Caldera , A. Trevor , K. Chiiba and N. Castagnoli, Jr . 1985. Studies on the 1-methyl-4-phenyl- 2, 3-dihydropyridinium species 2, 3-MPDP+, the monoamine oxidase catalyzed oxidation product of the nigro-striataltoxin 1-methyl-4-phenyl-1, 2, 3, 6-tetrahydropyridine (MPTP). J. Med. Chem. 28:1432–1436.

51.

Rabe, C. S. , E. Delorme and F. F. Weight . 1987. Muscarinic-stimulated neuro-transmitter release from PC 12 cells. J. Pharmacol. Exp. Ther. 243:534–541.

52.

Ramsay, R. R. , J. I. Salach , J. Dadgar and T. P. Singer . 1986. Inhibition of mitochondrial NADH dehydrogenaseby pyridine derivatives and its possible relation to experimental and idiopathic parkinsonism. Biochem. Biophys. Res. Commun. 135:269–275.

53.

Ramsay, R. R. , and T. P. Singer . 1986. Energy-dependent uptake on N-methyl-4-phenyl-pyridinium, the neurotoxic metabolite of 1-methyl-4-phenyl-1, 2, 3, 6-tetrahydropyridine, by mitochondria. J. Biol. Chem. 261:7585–7587.

54.

Ransom, B. R. , D. M. Kunis , I. Irwin and J. W. Langston . 1987. Astrocytes convert the parkinsonism inducing neurotoxin, MPTP, to its active metabolite, MPP+. Neurosci. Lett. 75:323–328.

55.

Reinchard, J. F. , E. J. DiLiberto , H. Viveros and A. J. Daniels . 1987. Subcellular compartmentalization of 1 -methyl-4-phenylpyridinium with catecholamines in adrenal medullary chromaffm vesicles may explain the lack of toxicity to adrenal chromaffm cells. Proc. Natl. Acad. Sci. USA 84:8160–8164.

56.

Reinchard, J. F. , D. B. Miller and J. P. Callaghan . 1988. The neurotoxicant MPTP (1-methyl-4-phenyl-1, 2, 3, 6-tetrahydropyridine) increases glial fibril-lary acidic protein and decreases dopamine levels of the mouse striatum: Evidence for glial response to injury. Neurosci. Lett. 95:246–251.

57.

Rollema, H. , G. Damsma , A. S. Horn , J. B. DeVries , and B. H. C. Westerink . 1986. Brain dialysis in conscious rats reveals an instantaneous massive release of sriatal dopamine in response to MPP+. Eur. J. Pharmacol. 126:345–346.

58.

Rollema, H. , W. G. Kuhr , G. Kranenborg , J. B. DeVries , and C. Vandenberg . 1988a. MPP+-induced efflux of dopamine and lactate from rat striatum have similar time courses as shown by in vivo brain dialysis. J. Pharmacol. Exp. Ther. 245:858–866.

59.

Rollema, H. , J. B. Vries , G. Damsma , B. H. C. Westerink , G. L. Kranenborg , W. G. Kuhr , and A. S. Horn . 1988b. The use of in vivo brain dialysis of dopamine, acetylcholine, amino acids and lactic acid in studies on the neurotoxin 1-methyl-4-phenyl-1, 2, 3, 6-tetrahydropyridine (MPTP). Toxicology 49:503–511.

60.

Ross R. A. , B. A. Spengler , and J. L. Biedler . 1983. Coordinate morphological and biochemical inter conversion of human neuroblastoma cells. JNCI 71:741–749.

61.

Rowles, T. K. , X. Song and M. Ehrich . 1995. Identification of endpoints affected by exposure of human neuroblastoma cells to neurotoxicants at concentrations below those that affect cell viability. In Vitro Toxicol. 8:1–11.

62.

Santiago, M. , A. Machado , and J. Cano . 1993. Regulation of prefrontal cortical dopamine release by dopamine receptor agonists and antagonists. Eur. J. Pharmacol. 239:83–91.

63.

Santiago, M. , H. Rollema , J. B. DeVries , and B. H. C. Westerink . 1991a. Acute effects of intranigral application of MPP+ on nigral and bilateral striatal release of dopamine simultaneously recorded by microdialysis. Brain Res. 538:226–230.

64.

Santiago, M. , B. H. C. Westerink , and H. Rollema . 1991b. Responsiveness of striatal dopamine release in awake animals after chronic 1-methyl-4-phenyl-pyridinium ion-induced lesions of the substantia nigra. J. Neurochem. 56: 1336–1342.

65.

Saporito, M. S. , R. E. Keikkila , S. K. Youngater , W. J. Nicklas and H. M. Geller . 1992. Dopaminergic neurotoxicity of 1-methyl-4-phenylpyridinium analogs in cultured neurons: Relationship to the dopamine uptake system and inhibition of mitochondria 1 respiration. J. Pharmacol. Exp. Ther. 260:1400–1409.

66.

Segel, I. H. 1976. Enzyme inhibition. In Biochemical calculations, 245–272. New York: John Wiley & Sons.

67.

Singer, T. P. , J. I. Salach , N. Castagnoli and A. Trevor . 1986. Interaction of the neurotoxic amine 1-methyl-4-phenyl-1, 2, 3, 6-tetrahydropyridine with mono-amine oxidase. Biochem. J. 235:785–789.

68.

Song, X., 1996. A study of the neurotoxicity of MPTP and analogs in human neuroblastoma SH-SY5Y cells. Ph.D. dissertation, Virginia Polytechnic Institute and State University, Blacksburg, VA.

69.

Song, X. , and M. Ehrich . 1998. Uptake and metabolism of MPTP and MPP+ in SH-SY5Y human neuroblastoma cells. In Vitro Toxicol. 11:3–14.

70.

Song, X. , M. Ehrich , D. Flaherty , Y. X. Wang , and N. Castagnoli . 1996. Biotrans-formation of the MPTP analog trans-1-methyl-4-[4-dimethylamino phenyl-ethenyl]-1, 2, 3, 6-tetrahydropyridine to a fluorescent pyridium metabolite by intact neuroblastoma cells. Toxicol. Appl. Pharmacol. 137:163–172.

71.

Song, X. , S. Perkins , B. S. Jortner , and M. Ehrich . 1997. Cytotoxic effects of MPTP on SH-SY5Y human neuroblastoma cells. Neuro Toxicology 18:341–354.

72.

Stern, G. 1990. The biochemistry of Parkinson's disease. In Parkinson's Disease, ed., G. Stern , 99–126. Baltimore, MD: The Johns Hopkins University Press.

73.

Uhl, G. R. , J. A. Javitch and S. H. Snyder . 1985. Normal MPTP binding in Parkinsonian substantia nigra: Evidence for extraneuronal toxin conversion in human brain. Lancet 1:956–957.

74.

Umemura, T. , M. Naoi , T. Takahaski , Y. Fukui , T. Yasue , M. Ohashi , and T. Nagatsu . 1990. Cytotoxic effect of 1-methyl-4-phenylpyridinium in human melanoma cell lines, HMV-11 and SK-MEL-44, is dependent on the melanin contents and caused by inhibition of mitochondrial electron transport. Bio-chem. Med. Metabol. Biol. 44:51–58.

75.

Vaughan, P. F. , D. F. Kaye , H. L. Reeve , S. G. Ball and C. Peers . 1993. Nicotinic receptor-mediated release of noradrenaline in the human neuroblastoma SH-SY5Y J. Neurochem. 60:2159–2166.

76.

Wall, S. J. , R. P. Yasuda , M. Li and B. B. Wolfe . 1991. Development of an anti-serum against m3 muscarinic receptors: Distribution of m3 receptor in rat tissues and clonal cell lines. Molec. Pharmacol. 40:783–789.

77.

Willets, J. M. , D. G. Lambert and H. R. Griffiths . 1993. Suitability of B65 and SH-SY5Y neuroblastoma cell as model for “in vitro” neurotoxicity testing. Biochem. Soc. Trans. 21:425s.