Neurochemical Research

, Volume 33, Issue 3, pp 401–411

The Role of 3-O-Methyldopa in the Side Effects of l-dopa

  • Eun-Sook Y. Lee
  • Hongtao Chen
  • Jennifer King
  • Clivel Charlton
Original Paper


Long-term treatment of l-dopa for Parkinson’s disease (PD) patients induces adverse effects, including dyskinesia, on–off and wearing-off symptoms. However, the cause of these side effects has not been established to date. In the present study, therefore, 3-O-methyldopa (3-OMD), which is a major metabolite of l-dopa, was tested to determine whether it plays a role in the aforementioned adverse effects. The effects of 3-OMD on the dopaminergic nervous system in the brain were investigated, by examining behavioral, biochemical, and cellular changes in male Sprague–Dawley rats and catecholamine-producing PC12 neuronal cells. The results revealed that the intracerebroventricular (icv) injection of 1 μmol of 3-OMD impaired locomotor activities by decreasing movement time (MT), total distance (TD), and the number of movement (NM) by 70, 74 and 61%, respectively. The biochemical analysis results showed that a single administration of 1 μmole of 3-OMD decreased the dopamine turnover rate (DOPAC/DA) by 40.0% in the rat striatum. 3-OMD inhibited dopamine transporter and uptake in rat brain striatal membranes and PC12 cells. The subacute administration of 3-OMD (5 days, icv) also significantly impaired the locomotor activities and catecholamine levels. 3-OMD induced cytotoxic effects via oxidative stress and decreased mitochondrial membrane potential in PC12 cells, indicating that 3-OMD can damage neuronal cells. Furthermore, 3-OMD potentiated l-dopa toxicity and these toxic effects induced by both 3-OMD and l-dopa were blocked by vitamin E (α-tocopherol) in PC12 cells, indicating that 3-OMD may increase the toxic effects of l-dopa to some extent by oxidative stress. Therefore, the present study reveals that 3-OMD accumulation from long-term l-dopa treatment may be involved in the adverse effects of l-dopa therapy. Moreover, l-dopa treatment might accelerate the progression of PD, at least in part, by 3-OMD.


l-dopa 3-O-methyldopa Parkinson’s disease Locomotor activity Oxidative stress 


  1. 1.
    Muenter MD, Sharpless NS, Tyce GM (1972) Plasma 3-0-methyldopa in l-dopa therapy of Parkinson’s disease. Mayo Clin Proc 47:389–395PubMedGoogle Scholar
  2. 2.
    Sharpless NS, Muenter MD, Tyce GM et al (1972) 3-methoxy-4-hydroxyphenylalanine (3-O-methyldopa) in plasma during oral l-dopa therapy of patients with Parkinson’s disease. Clin Chim Acta 37:359–369PubMedCrossRefGoogle Scholar
  3. 3.
    Gervas JJ, Muradas V, Bazan E et al (1983) Effects of 3-OM-dopa on monoamine metabolism in rat brain. Neurology 33:278–282PubMedGoogle Scholar
  4. 4.
    Tohgi H, Abe T, Takahashi S (1991b) Concentrations of tyrosine, l-dihydroxyphenylalanine, dopamine, and 3-O-methyldopa in the cerebrospinal fluid of Parkinson’s disease. Neurosci Lett 127:212–214PubMedCrossRefGoogle Scholar
  5. 5.
    Miller JW, Shukitt-Hale B, Villalobos-Molina R et al (1997) Effect of l-Dopa and the catechol-O-methyltransferase inhibitor Ro 41-0960 on sulfur amino acid metabolites in rats. Clin Neuropharmacol 20:55–66PubMedCrossRefGoogle Scholar
  6. 6.
    Kuruma I, Bartholini G, Tissot R et al (1971) The metabolism of l-3-O-methyldopa, a precursor of dopa in man. Clin Pharmacol Ther 12:678–82PubMedGoogle Scholar
  7. 7.
    Marion MH, Stocchi F, Quinn NP et al (1986) Repeated levodopa infusions in fluctuating Parkinson’s disease: clinical and pharmacokinetic data. Clin Neuropharmacol 9:165–181PubMedCrossRefGoogle Scholar
  8. 8.
    Marsden CD (1994) Problems with long-term levodopa therapy for Parkinson’s disease. Clin Neuropharmacol Suppl 2:S32–S44Google Scholar
  9. 9.
    Feuerstein C, Serre F, Gavend M et al (1977a) Plasma O-methyldopa in levodopa-induced dyskinesias. A bioclinical investigation. Acta Neurol Scand 56:508–524PubMedCrossRefGoogle Scholar
  10. 10.
    Feuerstein C, Tauche M, Serre F et al (1977b) Does O-methyl-dopa play a role in levodopa-induced dyskinesias. Acta Neurol Scand 56:79–82PubMedCrossRefGoogle Scholar
  11. 11.
    Mena MA, Muradas V, Bazan E et al (1987) Pharmacokinetics of l-dopa in patients with Parkinson’s disease. Adv Neurol 45:481–486PubMedGoogle Scholar
  12. 12.
    Blandini F, Nappi G, Fancellu R et al (2003) Modifications of plasma and platelet levels of l-DOPA and its direct metabolites during treatment with tolcapone or entacapone in patients with Parkinson’s disease. J Neural Transm 110:911–922PubMedCrossRefGoogle Scholar
  13. 13.
    Tohgi H, Abe T, Kikuchi T et al (1991a) The significance of 3-O-methyldopa concentrations in the cerebrospinal fluid in the pathogenesis of wearing-off phenomenon in Parkinson’s disease. Neurosci Lett 132:19–22PubMedCrossRefGoogle Scholar
  14. 14.
    Fabbrini G, Juncos JL, Mouradian MM et al (1987) 3-O-methyldopa and motor fluctuations in Parkinson’s disease. Neurology 37:856–859PubMedGoogle Scholar
  15. 15.
    Soares-da-Silva P, Parada A, Serrao P (2000) The O-methylated derivative of l-DOPA, 3-O-methyl-l-DOPA, fails to inhibit neuronal and non-neuronal aromatic l-amino acid decarboxylase. Brain Res 863:293–297PubMedCrossRefGoogle Scholar
  16. 16.
    Reches A, Fahn S (1982) 3-O-methyldopa blocks dopa metabolism in rat corpus striatum. Ann Neurol 12:267–271PubMedCrossRefGoogle Scholar
  17. 17.
    Benetello P, Furlanut M, Fortunato M et al (1997) Levodopa and 3-O-methyldopa in cerebrospinal fluid after levodopa-carbidopa association. Pharmacol Res 35:313–315PubMedCrossRefGoogle Scholar
  18. 18.
    Cheng H, Gomes-Trolin C, Aquilonius SM et al (1997) Levels of l-methionine S-adenosyltransferase activity in erythrocytes and concentrations of S-adenosylmethionine and S-adenosylhomocysteine in whole blood of patients with Parkinson’s disease. Exp Neurol 145:580–585PubMedCrossRefGoogle Scholar
  19. 19.
    Zhao WQ, Latinwo L, Liu XX et al (2001) l-dopa upregulates the expression and activities of methionine adenosyl transferase and catechol-O-methyltransferase. Exp Neurol 171:127–138PubMedCrossRefGoogle Scholar
  20. 20.
    Chang WY, Webster RA (1995) Effects of 3-O-methyl on l-dopa-facilitated synthesis and efflux of dopamine from rat striatal slices. Br J Pharmacol 116:2637–2640PubMedGoogle Scholar
  21. 21.
    Mena MA, Pardo B, Paino CL et al (1993) Levodopa toxicity in foetal rat midbrain neurons in culture: modulation by ascorbic acid. Neuroreport 4:438–440PubMedCrossRefGoogle Scholar
  22. 22.
    Mena MA, Casarejos MJ, Carazo A et al (1996) Glia conditioned medium protects fetal rat midbrain neurones in culture from l-DOPA toxicity. Neuroreport 7:441–445PubMedCrossRefGoogle Scholar
  23. 23.
    Datla KP, Blunt SB, Dexter DT (2001) Chronic l-DOPA administration is not toxic to the remaining dopaminergic nigrostriatal neurons, but instead may promote their functional recovery, in rats with partial 6-OHDA or FeCl(3) nigrostriatal lesions. Mov Disord 16:424–434PubMedCrossRefGoogle Scholar
  24. 24.
    Melamed E, Offen D, Shirvan A et al (1998) Levodopa toxicity and apoptosis. Ann Neurol 44:S149–S154PubMedGoogle Scholar
  25. 25.
    Murer MG, Dziewczapolski G, Menalled LB et al (1998) Chronic levodopa is not toxic for remaining dopamine neurons, but instead promotes their recovery, in rats with moderate nigrostriatal lesions. Ann Neurol 43:561–575PubMedCrossRefGoogle Scholar
  26. 26.
    Walkinshaw G, Waters CM (1995) Induction of apoptosis in catecholaminergic PC12 cells by l-DOPA. Implications for the treatment of Parkinson’s disease. J Clin Invest 95:2458–2464PubMedGoogle Scholar
  27. 27.
    Müller T, Woitalla D, Fowler B et al (2002) 3-OMD and homocysteine plasma levels in parkinsonian patients. J Neural Transm 109:175–179PubMedCrossRefGoogle Scholar
  28. 28.
    Müller T, Woitalla D, Hauptmann B et al (2001) Decrease of methionine and S-adenosylmethionine and increase of homocysteine in treated patients with Parkinson’s disease. Neurosci Lett 308:54–56PubMedCrossRefGoogle Scholar
  29. 29.
    Lee ES, Chen H, Soliman KF et al (2005) Effects of homocysteine on the dopaminergic system and behavior in rodents. Neurotoxicology 26:361–371PubMedCrossRefGoogle Scholar
  30. 30.
    Spencer JP, Jenner A, Aruoma OI et al (1994) Intense oxidative DNA damage promoted by l-dopa and its metabolites. Implications for neurodegenerative disease. FEBS Lett 353:246–250PubMedCrossRefGoogle Scholar
  31. 31.
    Martignoni E, Blandini F, Godi L et al (1999) Peripheral markers of oxidative stress in Parkinson’s disease. The role of l-DOPA. Free Radic Biol Med 27:428–437CrossRefGoogle Scholar
  32. 32.
    Blandini F, Martignoni E, Ricotti R et al (1999) Determination of hydroxyl free radical formation in human platelets using high-performance liquid chromatography with electrochemical detection. J Chromatogr B Biomed Sci Appl 732:213–220PubMedCrossRefGoogle Scholar
  33. 33.
    Lee JJ, Kim YM, Yin SY et al (2003) Aggravation of l-DOPA-induced neurotoxicity by tetrahydropapaveroline in PC12 cells. Biochem Pharmacol 66:1787–1795PubMedCrossRefGoogle Scholar
  34. 34.
    Lowry CA, Renner KJ, Moore FL (1996) Catecholamines and indoleamines in the central nervous system of a urodele amphibian: a microdissection study with emphasis on the distribution of epinephrine. Brain Behav Evol 48:70–93PubMedCrossRefGoogle Scholar
  35. 35.
    Billard W, Ruperto V, Crosby G et al (1984) Characterization of the binding of [3H]SCH 23390, a selective D1 receptor antagonist ligand, in rat striatum. Life Sci 35:1885–1893PubMedCrossRefGoogle Scholar
  36. 36.
    Woodgate A, MacGibbon G, Walton M et al (1999) The toxicity of 6-hydroxydopamine on PC12 and P19 cells. Brain Res Mol Brain Res 69:84–92PubMedCrossRefGoogle Scholar
  37. 37.
    Fu W, Luo H, Parthasarathy S et al (1998) Catecholamines potentiate amyloid beta-peptide neurotoxicity: involvement of oxidative stress, mitochondrial dysfunction, and perturbed calcium homeostasis. Neurobiol Dis 5:229–243PubMedCrossRefGoogle Scholar
  38. 38.
    Palmeira CM, Moreno AJ, Madeira VM et al (1996) Continuous monitoring of mitochondrial membrane potential in hepatocyte cell suspensions. J Pharmacol Toxicol Methods 35:35–43PubMedCrossRefGoogle Scholar
  39. 39.
    Lee CS, Han JH, Jang YY et al (2002) Differential effect of catecholamines and MPP+ on membrane permeability in brain mitochondria and cell viability in PC12 cells. Neurochem Int 40:361–369PubMedCrossRefGoogle Scholar
  40. 40.
    Reilly DK, Rivera-Calimlim L, Van Dyke D (1980) Catechol-O-methyltransferase activity: a determinant of levodopa response. Clin Pharmacol Ther 28:278–286PubMedCrossRefGoogle Scholar
  41. 41.
    Rivera-Calimlim L, Tandon D, Anderson F et al (1977) The clinical picture and plasma levodopa metabolite profile of parkinsonian nonresponders. Treatment with levodopa and decarboxylase inhibitor. Arch Neurol 34:228–232PubMedGoogle Scholar
  42. 42.
    Gomes P, Soares-da-Silva P (1999) Interaction between l-DOPA and 3-O-methyl-l-DOPA for transport in immortalised rat capillary cerebral endothelial cells. Neuropharmacology 38:1371–1380PubMedCrossRefGoogle Scholar
  43. 43.
    Himori N, Tanaka Y, Kurasawa M et al (1994) 3-O-methyldopa attenuates the effects of Madopar on the haloperidol-induced cataleptic behavior and the locomotor activity in the mouse. Pharmacology 48:226–233PubMedCrossRefGoogle Scholar
  44. 44.
    Greene LA, Rein G (1977) Release of (3H)norepinephrine from a clonal line of pheochromocytoma cells (PC12) by nicotinic cholinergic stimulation. Brain Res 138:521–528PubMedCrossRefGoogle Scholar
  45. 45.
    Schubert D, Klier FG (1977) Storage and release of acetylcholine by a clonal cell line. Proc Natl Acad Sci USA 74:5184–5188PubMedCrossRefGoogle Scholar
  46. 46.
    Rebois RV, Reynolds EE, Toll L et al (1980) Storage of dopamine and acetylcholine in granules of PC12, a clonal pheochromocytoma cell line. Biochemistry 19:1240–1248PubMedCrossRefGoogle Scholar
  47. 47.
    Greene LA, Tischler AS (1976) Establishment of a noradrenergic clonal line of rat adrenal pheochromocytoma cells which respond to nerve growth factor. Proc Natl Acad Sci USA 73:2424–2428PubMedCrossRefGoogle Scholar
  48. 48.
    Basma AN, Morris EJ, Nicklas WJ, Geller HM (1995) l-dopa cytotoxicity to PC12 cells in culture is via its autoxidation. J Neurochem 64:825–832PubMedCrossRefGoogle Scholar
  49. 49.
    Cobuzzi RJ, Neafsey EJ, Collins MA (1994) Differential cytotoxicities of N-methyl-beta-carbolinium analogues of MPP+ in PC12 cells: insights into potential neurotoxicants in Parkinson’s disease. J Neurochem 62:1503–1510PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2007

Authors and Affiliations

  • Eun-Sook Y. Lee
    • 1
  • Hongtao Chen
    • 1
  • Jennifer King
    • 1
  • Clivel Charlton
    • 1
  1. 1.Department of Neurobiology and NeurotoxicologyMeharry Medical CollegeNashvilleUSA

Personalised recommendations