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. 1992 Oct 1;89(19):9074–9078. doi: 10.1073/pnas.89.19.9074

Gene transfer of a reserpine-sensitive mechanism of resistance to N-methyl-4-phenylpyridinium.

Y Liu 1, A Roghani 1, R H Edwards 1
PMCID: PMC50067  PMID: 1409604

Abstract

The toxin N-methyl-1,2,3,6-tetrahydropyridine produces a model of neural degeneration very similar to idiopathic Parkinson disease. To understand the cellular mechanisms that modulate susceptibility to its active metabolite N-methyl-4-phenylpyridinium (MPP+), we have transfected a cDNA expression library from the relatively MPP(+)-resistant rat pheochromocytoma PC12 cells into MPP(+)-sensitive Chinese hamster ovary (CHO) fibroblasts. Selection of the stable transformants in high concentrations of MPP+ has yielded a clone extremely resistant to the toxin. Reserpine reverses the resistance to MPP+, suggesting that a transport activity protects against this form of toxicity, perhaps by sequestering the toxin within an intracellular compartment. In support of this hypothesis, dopamine loaded into the CHO transformant shows a localized distribution that is distinct from the pattern observed in wild-type cells and is also reversed by reserpine.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Aruffo A., Seed B. Molecular cloning of a CD28 cDNA by a high-efficiency COS cell expression system. Proc Natl Acad Sci U S A. 1987 Dec;84(23):8573–8577. doi: 10.1073/pnas.84.23.8573. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Aviv H., Leder P. Purification of biologically active globin messenger RNA by chromatography on oligothymidylic acid-cellulose. Proc Natl Acad Sci U S A. 1972 Jun;69(6):1408–1412. doi: 10.1073/pnas.69.6.1408. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Chen C., Okayama H. High-efficiency transformation of mammalian cells by plasmid DNA. Mol Cell Biol. 1987 Aug;7(8):2745–2752. doi: 10.1128/mcb.7.8.2745. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Chirgwin J. M., Przybyla A. E., MacDonald R. J., Rutter W. J. Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease. Biochemistry. 1979 Nov 27;18(24):5294–5299. doi: 10.1021/bi00591a005. [DOI] [PubMed] [Google Scholar]
  5. Cohen G. Monoamine oxidase and oxidative stress at dopaminergic synapses. J Neural Transm Suppl. 1990;32:229–238. doi: 10.1007/978-3-7091-9113-2_33. [DOI] [PubMed] [Google Scholar]
  6. Daniels A. J., Reinhard J. F., Jr Energy-driven uptake of the neurotoxin 1-methyl-4-phenylpyridinium into chromaffin granules via the catecholamine transporter. J Biol Chem. 1988 Apr 15;263(11):5034–5036. [PubMed] [Google Scholar]
  7. De la Torre J. C. An improved approach to histofluorescence using the SPG method for tissue monoamines. J Neurosci Methods. 1980 Oct;3(1):1–5. doi: 10.1016/0165-0270(80)90029-1. [DOI] [PubMed] [Google Scholar]
  8. Denton T., Howard B. D. A dopaminergic cell line variant resistant to the neurotoxin 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine. J Neurochem. 1987 Aug;49(2):622–630. doi: 10.1111/j.1471-4159.1987.tb02909.x. [DOI] [PubMed] [Google Scholar]
  9. Dower W. J., Miller J. F., Ragsdale C. W. High efficiency transformation of E. coli by high voltage electroporation. Nucleic Acids Res. 1988 Jul 11;16(13):6127–6145. doi: 10.1093/nar/16.13.6127. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Greene L. A., Rein G. Release, storage and uptake of catecholamines by a clonal cell line of nerve growth factor (NGF) responsive pheo-chromocytoma cells. Brain Res. 1977 Jul 1;129(2):247–263. doi: 10.1016/0006-8993(77)90005-1. [DOI] [PubMed] [Google Scholar]
  11. Gubler U., Hoffman B. J. A simple and very efficient method for generating cDNA libraries. Gene. 1983 Nov;25(2-3):263–269. doi: 10.1016/0378-1119(83)90230-5. [DOI] [PubMed] [Google Scholar]
  12. Hansen M. B., Nielsen S. E., Berg K. Re-examination and further development of a precise and rapid dye method for measuring cell growth/cell kill. J Immunol Methods. 1989 May 12;119(2):203–210. doi: 10.1016/0022-1759(89)90397-9. [DOI] [PubMed] [Google Scholar]
  13. Heikkila R. E., Manzino L., Cabbat F. S., Duvoisin R. C. Protection against the dopaminergic neurotoxicity of 1-methyl-4-phenyl-1,2,5,6-tetrahydropyridine by monoamine oxidase inhibitors. Nature. 1984 Oct 4;311(5985):467–469. doi: 10.1038/311467a0. [DOI] [PubMed] [Google Scholar]
  14. Hyman C., Hofer M., Barde Y. A., Juhasz M., Yancopoulos G. D., Squinto S. P., Lindsay R. M. BDNF is a neurotrophic factor for dopaminergic neurons of the substantia nigra. Nature. 1991 Mar 21;350(6315):230–232. doi: 10.1038/350230a0. [DOI] [PubMed] [Google Scholar]
  15. Javitch J. A., D'Amato R. J., Strittmatter S. M., Snyder S. H. 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 U S A. 1985 Apr;82(7):2173–2177. doi: 10.1073/pnas.82.7.2173. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Kilty J. E., Lorang D., Amara S. G. Cloning and expression of a cocaine-sensitive rat dopamine transporter. Science. 1991 Oct 25;254(5031):578–579. doi: 10.1126/science.1948035. [DOI] [PubMed] [Google Scholar]
  17. Kish P. E., Fischer-Bovenkerk C., Ueda T. Active transport of gamma-aminobutyric acid and glycine into synaptic vesicles. Proc Natl Acad Sci U S A. 1989 May;86(10):3877–3881. doi: 10.1073/pnas.86.10.3877. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Knigge K. M., Hoffman G., Scott D. E., Sladek J. R., Jr Identification of catecholamine and luteinizing hormone-releasing hormone (LHRH)-containing neurons in primary cultures of dispersed cells of the basal hypothalamus. Brain Res. 1977 Jan 28;120(3):393–405. doi: 10.1016/0006-8993(77)90394-8. [DOI] [PubMed] [Google Scholar]
  19. Krueger M. J., Singer T. P., Casida J. E., Ramsay R. R. Evidence that the blockade of mitochondrial respiration by the neurotoxin 1-methyl-4-phenylpyridinium (MPP+) involves binding at the same site as the respiratory inhibitor, rotenone. Biochem Biophys Res Commun. 1990 May 31;169(1):123–128. doi: 10.1016/0006-291x(90)91442-u. [DOI] [PubMed] [Google Scholar]
  20. Langston J. W., Ballard P., Tetrud J. W., Irwin I. Chronic Parkinsonism in humans due to a product of meperidine-analog synthesis. Science. 1983 Feb 25;219(4587):979–980. doi: 10.1126/science.6823561. [DOI] [PubMed] [Google Scholar]
  21. Langston J. W., Irwin I., Langston E. B., Forno L. S. Pargyline prevents MPTP-induced parkinsonism in primates. Science. 1984 Sep 28;225(4669):1480–1482. doi: 10.1126/science.6332378. [DOI] [PubMed] [Google Scholar]
  22. Markey S. P., Johannessen J. N., Chiueh C. C., Burns R. S., Herkenham M. A. Intraneuronal generation of a pyridinium metabolite may cause drug-induced parkinsonism. Nature. 1984 Oct 4;311(5985):464–467. doi: 10.1038/311464a0. [DOI] [PubMed] [Google Scholar]
  23. Mizuno Y., Ohta S., Tanaka M., Takamiya S., Suzuki K., Sato T., Oya H., Ozawa T., Kagawa Y. Deficiencies in complex I subunits of the respiratory chain in Parkinson's disease. Biochem Biophys Res Commun. 1989 Sep 29;163(3):1450–1455. doi: 10.1016/0006-291x(89)91141-8. [DOI] [PubMed] [Google Scholar]
  24. Pacholczyk T., Blakely R. D., Amara S. G. Expression cloning of a cocaine- and antidepressant-sensitive human noradrenaline transporter. Nature. 1991 Mar 28;350(6316):350–354. doi: 10.1038/350350a0. [DOI] [PubMed] [Google Scholar]
  25. Parker W. D., Jr, Boyson S. J., Parks J. K. Abnormalities of the electron transport chain in idiopathic Parkinson's disease. Ann Neurol. 1989 Dec;26(6):719–723. doi: 10.1002/ana.410260606. [DOI] [PubMed] [Google Scholar]
  26. Ramsay R. R., Krueger M. J., Youngster S. K., Gluck M. R., Casida J. E., Singer T. P. Interaction of 1-methyl-4-phenylpyridinium ion (MPP+) and its analogs with the rotenone/piericidin binding site of NADH dehydrogenase. J Neurochem. 1991 Apr;56(4):1184–1190. doi: 10.1111/j.1471-4159.1991.tb11409.x. [DOI] [PubMed] [Google Scholar]
  27. Ramsay R. R., Singer T. P. 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. 1986 Jun 15;261(17):7585–7587. [PubMed] [Google Scholar]
  28. Reinhard J. F., Jr, Carmichael S. W., Daniels A. J. Mechanisms of toxicity and cellular resistance to 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine and 1-methyl-4-phenylpyridinium in adrenomedullary chromaffin cell cultures. J Neurochem. 1990 Jul;55(1):311–320. doi: 10.1111/j.1471-4159.1990.tb08853.x. [DOI] [PubMed] [Google Scholar]
  29. Reinhard J. F., Jr, Diliberto E. J., Jr, Viveros O. H., Daniels A. J. Subcellular compartmentalization of 1-methyl-4-phenylpyridinium with catecholamines in adrenal medullary chromaffin vesicles may explain the lack of toxicity to adrenal chromaffin cells. Proc Natl Acad Sci U S A. 1987 Nov;84(22):8160–8164. doi: 10.1073/pnas.84.22.8160. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Rudnick G. ATP-driven H+ pumping into intracellular organelles. Annu Rev Physiol. 1986;48:403–413. doi: 10.1146/annurev.ph.48.030186.002155. [DOI] [PubMed] [Google Scholar]
  31. Scherman D., Henry J. P. Reserpine binding to bovine chromaffin granule membranes. Characterization and comparison with dihydrotetrabenazine binding. Mol Pharmacol. 1984 Jan;25(1):113–122. [PubMed] [Google Scholar]
  32. Shimada S., Kitayama S., Lin C. L., Patel A., Nanthakumar E., Gregor P., Kuhar M., Uhl G. Cloning and expression of a cocaine-sensitive dopamine transporter complementary DNA. Science. 1991 Oct 25;254(5031):576–578. doi: 10.1126/science.1948034. [DOI] [PubMed] [Google Scholar]
  33. Shoffner J. M., Watts R. L., Juncos J. L., Torroni A., Wallace D. C. Mitochondrial oxidative phosphorylation defects in Parkinson's disease. Ann Neurol. 1991 Sep;30(3):332–339. doi: 10.1002/ana.410300304. [DOI] [PubMed] [Google Scholar]
  34. Stern-Bach Y., Greenberg-Ofrath N., Flechner I., Schuldiner S. Identification and purification of a functional amine transporter from bovine chromaffin granules. J Biol Chem. 1990 Mar 5;265(7):3961–3966. [PubMed] [Google Scholar]
  35. Turski L., Bressler K., Rettig K. J., Löschmann P. A., Wachtel H. Protection of substantia nigra from MPP+ neurotoxicity by N-methyl-D-aspartate antagonists. Nature. 1991 Jan 31;349(6308):414–418. doi: 10.1038/349414a0. [DOI] [PubMed] [Google Scholar]

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