Neuromelanin synthesis in rat and human substantia nigra

  • J. M. Rabey
  • F. Hefti
Original Papers


A relation between neuromelanin synthesis and vulnerability of dopaminergic neurons is suggested by the fact that heavily pigmented cells are preferentially lost in aging and Parkinson's disease and that the dopaminergic neurotoxin MPP+ (1-methyl-4-phenyl-pyridine) binds to neuromelanin. To elucidate the mechanism of neuromelanin synthesis, we studied the formation of melanin in homogenates of human and rat substantia nigra tissue “in vitro”. It was found that enzymatic processes accounted for 70% and 90% of the melanin formation in homogenates of human and rat tissue, respectively. The enzymatic synthesis was due to the activity of monoamine oxidase (MAO), since it was prevented by selective inhibitors of this enzyme. Both MPTP (1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine) and MPP+ inhibited melanin formation, probably due to their ability to inhibit MAO. No evidence was found for involvement of cytochrome P-450 monooxigenases, which have been postulated to exist in central catecholaminergic neurons. Proadifen reduced melanin formation, not necessarily because it is an inhibitor of P-450 monooxigenases, but rather as it is also a potent inhibitor of MAO. Some antioxidants like ascorbic acid, but not agents destroying hydrogen peroxide, inhibited melanin formation. The findings suggest that the formation of neuromelanin in the substantia nigra involves MAO and non-enzymatic oxidative processes.


MPTP MPP+ neuromelanin Parkinson's disease 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Barden H (1969) The histochemical relationship of neuromelanin and lipofucsin. J Neuropathol Exp Neurol 28: 419–441PubMedGoogle Scholar
  2. Bazelon M, Fenichel GM, Randall J (1967) Studies on neuromelanin. I. A melanin system in the human adult brainstem. Neurology 17: 512–518PubMedGoogle Scholar
  3. Blaschko H, Hellmann K (1953) Pigment formation from tryptamine and 5-hydroxytryptamine in tissue: a contribution to the histochemistry of amine oxidase. J Physiol 122: 419–427PubMedGoogle Scholar
  4. Bochetta A, Piccardi MP, Del Zompo M, Pintus S, Corsini GU (1985) 1-methyl-4-phenyl-1,2,5,6-tetrahydropyridine: correspondence of its binding sites to monoamine oxidase in rat brain and inhibition of dopamine oxidative deamination in vivo and in vitro. J Neurochem 45: 673–676PubMedGoogle Scholar
  5. Bogerts B (1981) A brainstem atlas of catecholaminergic neurons in man using melanin as a natural marker. J Comp Neurol 197: 63–80CrossRefPubMedGoogle Scholar
  6. Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72: 248–254PubMedGoogle Scholar
  7. Cohen G, Pasik P, Cohen B, Leist A, Mytilineou C, Yahr MD (1985) Pargyline and deprenyl prevent the neurotoxicity of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) in monkeys. Eur J Pharmacol 106: 209–210CrossRefGoogle Scholar
  8. D'Amato RJ, Lipman ZP, Snyder SH (1986) Selectivity of the parkinsonian neurotoxin MPTP: toxic metabolite MPP binds to neuromelanin. Science 231: 987–989PubMedGoogle Scholar
  9. Das KC, Abramson MB, Katzman R (1978) Neuronal pigments: spectroscopic characterization of human brain melanin. J Neurochem 30: 601–605PubMedGoogle Scholar
  10. Del Zompo M, Piccardi MP, Bernardi F, Bonuccelli U, Corsini GU (1986) Involvement of monoamine oxidase enzymes in the action of 1-methyl-4-phenyl-1,2,5,6-tetrahydropyridine, a selective neurotoxin in the Squirrel monkey: binding and biochemical studies. Brain Res 378: 320–324CrossRefPubMedGoogle Scholar
  11. DeMattei M, Levi AC, Fariello RG (1986) Neuromelanic pigment in substantia nigra neurons of rats and dogs. Neurosci Lett 72: 37–42CrossRefPubMedGoogle Scholar
  12. Fellman JH (1958) Epinephrine metabolites and pigmentation in the central nervous system in a case of phenylpyruvic oligophrenia. J Neurol Neurosurg Psychiatry 21: 58–62PubMedGoogle Scholar
  13. Foley JM, Baxter D (1958) On the nature of pigment granules in the cells of the locus ceruleus and substantia nigra. J Neuropathol Exp Neurol 17: 586–598PubMedGoogle Scholar
  14. Fuller RW, Steranka LR (1985) Central and peripheral catecholamine depletion by 1-methyl-4-phenyl-tetrahydropyridine (MPTP) in rodents. Life Sci 36: 243–247PubMedGoogle Scholar
  15. Graham DG, Tye RW, Vogel FS (1977) Inhibition of DNA polymerase from L 1210 murine leukemia by a sulfhydryl reagent from Agaricus bisporus. Cancer Res 37: 436–439PubMedGoogle Scholar
  16. Graham DG (1978a) Oxidative pathways for catecholamines in the genesis of neuromelanin and cytotoxic quinones. Mol Pharmacol 14: 633–643PubMedGoogle Scholar
  17. Graham DG, Tiffany SM, Bell WR Jr (1978b) Autooxidation versus covalent binding of quinones as the mechanism of toxicity of dopamine, 6-hydroxydopamine and related compounds towards C 1300 neuroblastoma cells in vitro. Mol Pharmacol 14: 644–653PubMedGoogle Scholar
  18. Graham DG (1979) On the origin and significance of neuromelanin. Arch Pathol Lab Med 103: 359–362PubMedGoogle Scholar
  19. Grisham MB, Perez VJ, Everse J (1987) Neuromelanogenic and cytotoxic properties of canine brainstem peroxidase. J Neurochem 48: 876–882PubMedGoogle Scholar
  20. Haglund L, Kohler C, Haaparanta T, Goldstein M, Gustafsson JA (1984) Presence of NADPH-cytochrome P-450 reductase in central catecholaminergic neurons. Nature 107: 259–262Google Scholar
  21. Heikkila RE, Hess A, Duvoisin RC (1984) Dopaminergic neurotoxicity of 1-methyl-4-phenyl-1,2,5,6-tetrahydropyridine in mice. Science 224: 1451–1453PubMedGoogle Scholar
  22. Hirsch E, Graybiel AM, Agid YA (1988) Melanized dopaminergic neurons are differentially susceptible to degeneration in Parkinson's disease. Nature 334: 345–348CrossRefPubMedGoogle Scholar
  23. Knoll J (1983) Deprenyl (selegiline): the history of its development and pharmacological action. Acta Neurol Scand 95: 57–80Google Scholar
  24. Kopin IJ, Burns SR, Chiueh CC, Markey SP (1986) MPTP induced Parkinsonian syndromes in humans and animals. In: Fisher A, Hanin I, Lachman CH (eds) Alzheimer and Parkinson's disease. Strategies for research and development. Plenum Press, New York (Adv Behav Biol, vol 29, pp 519–530)Google Scholar
  25. Lichtensteiger W (1971) Effect of electrical stimulation on the fluorescence intensity of catecholamine-containing tuberal nerve cells. J Physiol 218: 63–84PubMedGoogle Scholar
  26. Lichtensteiger W, Felix D, Lienhart R, Hefti F (1976) A quantitative correlation between single unit activity and fluorescence intensity of dopamine neurons in zona compacta of substantia nigra, as demonstrated under the influence of nicotine and physostigmine. Brain Res 117: 85–103CrossRefPubMedGoogle Scholar
  27. Lichtensteiger W, Felix D, Hefti F (1979) Spike activity and histofluorescence correlated in the giant dopamine neuron of Planorbis corneus. Brain Res 170: 231–245CrossRefPubMedGoogle Scholar
  28. Lillie RD (1957) Metal reduction reactions of the melanins: histochemical studies. J Histochem Cytochem 5: 325–333PubMedGoogle Scholar
  29. Lyden A, Bondesson V, Larsson BS, Lindquist NG (1983) Melanin affinity of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine, an inducer of chronic parkinsonism in humans. Acta Pharmacol Toxicol (Copenh) 53: 429–432Google Scholar
  30. Maker HS, Weiss C, Silides DJ, Cohen G (1981) Coupling of dopamine oxidation (monoamine oxidase activity) to glutathione oxidation via the generation of hydrogen peroxide in rat brain homogenates. J Neurochem 36: 589–593PubMedGoogle Scholar
  31. Mann DMA, Yates PO (1974) Lipoprotein pigments. Their relationship to aging in the human nervous system. II. The melanin content of pigmented nerve cells. Brain 97: 489–498PubMedGoogle Scholar
  32. Mann DMA, Yates PO (1977) Neuromelanin and RNA in cells of substantia nigra. J Neuropathol Exp Neurol 36: 379–383PubMedGoogle Scholar
  33. Mann DMA, Yates PO (1983) Possible role of neuromelanin in the pathogenesis of Parkinson disease. Mechanism of aging and development, vol 21. Elsevier, pp 193–203CrossRefGoogle Scholar
  34. Marsden CD (1983) Neuromelanin and Parkinson's disease. J Neural Transm 19: 121–141Google Scholar
  35. Mayer SE, Melmon KL, Gilman AG (1980) The dynamics of drug absorption, distribution and elimination. In: Goodman Gilman A, Goodman LS, Gilman A (eds) The pharmacological basis of therapeutics. McMillan, New York, pp 1–27Google Scholar
  36. Melamed E, Rosenthal J, Cohen O, Uzzan A, Globus M (1985) Amphetamine but not reserpine protects mice against dopaminergic toxicity of MPTP. Neuropharmacology 24: 923–925CrossRefPubMedGoogle Scholar
  37. Parsons B, Rainbow TC (1984) High affinity binding sites for3H-MPTP may correspond to monoamine oxidase. Eur J Pharmacol 102: 375–377CrossRefPubMedGoogle Scholar
  38. Perry TL, Yong VW, Clavier RM, Jones K, Wright JM, Foulks JG, Wall RA (1985) Partial protection from the dopaminergic neurotoxin N-methyl-4-phenyl-1,2,3,6-tetrahydropyridine by four different antioxidants in the mouse. Neurosci Lett 60: 109–114CrossRefPubMedGoogle Scholar
  39. Pomerantz SH (1958) Separation, purification and properties of two tyrosinase from Hamster melanoma. J Neuropathol Exp Neurol 17: 586–598PubMedGoogle Scholar
  40. Prota G (1980) Recent advances in the chemistry of melanogenesis in mammals. J Invest Dermatol 75: 122–127CrossRefPubMedGoogle Scholar
  41. Rainbow TC, Parsons B, Wieczorek CM, Manaker S (1985) Localization in rat brain of binding sites for parkinsonian toxin MPTP: similarities with3H-pargyline binding to monoamine oxidase. Brain Res 330: 337–342CrossRefPubMedGoogle Scholar
  42. Rodgers AD, Curzon G (1975) Melanin formation by human brain in vitro. J Neurochem 24: 1123–1129PubMedGoogle Scholar
  43. Roth JA, Feor K (1978) Deamination of dopamine and its 3-o-methylated derivative by human brain monoamine oxidase. Biochem Pharmacol 27: 1606–1608CrossRefPubMedGoogle Scholar
  44. Schmidt DJ, Bruckwick E, Lovenberg W (1985) Lack of evidence supporting a role for dopamine in 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine neurotoxicity. Eur J Pharmacol 113: 149–150CrossRefPubMedGoogle Scholar
  45. Singer T, Salach J, Crabtee D (1985) Reversible inhibition and mechanism based irreversible inactivation of monoamine oxidases by 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP). Biochem Biophys Res Commun 127: 707–712PubMedGoogle Scholar
  46. Smith MT, Ekstrom G, Sandy MS, DiMonte D (1987) Current concepts in molecular mechanisms of MPTP-induced toxicity. VI. Studies on the mechanism of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine cytotoxicity in isolated hepatocytes. Life Sci 40:741–748CrossRefPubMedGoogle Scholar
  47. Snyder SH, D'Amato R (1986) MPTP: a neurotoxin relevant to the pathophysiology of Parkinson's disease. Neurology 36: 250–258PubMedGoogle Scholar
  48. Tiffany SM, Graham DG, Vogel FS, Cass MW, Jeffs AB (1978) Investigation of structure-function relationships of cytotoxic quinones of natural and synthetic origin. Cancer Res 38: 3230–3235PubMedGoogle Scholar
  49. Van Woert MH, Prasad KN, Borg DC (1967) Spectroscopic studies of substantia nigra pigment in human subjects. J Neurochem 14: 707–716PubMedGoogle Scholar
  50. Wagner GC, Carelli RM, Jarvis MF (1986) Ascorbic acid reduces the dopamine depletion induced by methamphetamine and the 1-methyl-4-phenyl pyridinium. Neuropharmacology 25: 559–561CrossRefPubMedGoogle Scholar
  51. Westlund KN, Denney RM, Kochersperger LM, Rose RM, Abell CW (1985) Distinct monoamine oxidase A and B populations in primate brain. Science 230: 181–183PubMedGoogle Scholar
  52. Wu EY, Chiba K, Trevor AJ, Castagnoli N (1986) Interactions of the 1-methyl-4-phenyl-2,3-dihydropyridinium species wtih synthetic dopamine-melanin. Life Sci 39: 1695–1700CrossRefPubMedGoogle Scholar
  53. Yong VW, Perry TL (1986) Monoamine oxidase B, smoking and Parkison's disease. J Neurol Sci 72: 265–272CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag 1990

Authors and Affiliations

  • J. M. Rabey
    • 1
  • F. Hefti
    • 1
  1. 1.Department of NeurologyUniversity of Miami School of MedicineMiamiU.S.A.

Personalised recommendations