Abstract
Aim:
To study the role of dopamine (DA) in rotenone-induced neurotoxicity in PC12 cells.
Methods:
Cell viability was assessed by detecting the leakage of lactate dehydrogenase (LDH) into the medium. Apoptosis rate was measured by flow cytometry. Caspase-3-like activity was measured by fluorescence assay using the probe Ac-DEVD-AMC. The level of intracellular hydrogen peroxide and other peroxides in PC12 cells were quantified by loading cells with 2′-7′-Dichlorodihydrofluorescein diacetate (DCFH-DA) in fluorescence assay. Lactic acid was measured spectrophotometrically. The DA levels in PC12 cells were determined by HPLC-ECD.
Results:
A 48-h incubation of PC12 cells with rotenone caused an apoptotic cell death and elevated intracellular reactive oxygen species (ROS) and lactic acid accumulation. Intracellular DA depletion with reserpine significantly attenuated rotenone-induced ROS accumulation and apoptotic cell death. No change was found in rotenone-induced ROS accumulation when cells were co-treated with deprenyl. Brief treatment with reserpine at the end of rotenone treatment had no effect on rotenone-induced neurotoxicity. However, when cells were first incubated with deprenyl, a monoamine oxidase-B inhibitor for 30 min then co-incubated with rotenone plus deprenyl, a brief treatment with reserpine enhanced cell injury.
Conclusion:
Rotenone-induced apoptosis in PC12 cells was mediated by intracellular dopamine oxidation.
Similar content being viewed by others
Article PDF
References
Kitada T, Asakawa S, Hattori N, Matsumine H, Yamamura Y, Minoshima S, et al. Mutations in the parkin gene cause autosomal recessive juvenile parkinsonism. Nature 1998; 392: 605–8.
Leroy E, Boyer R, Auburger G, Leube B, Ulm G, Mezey E, et al. The ubiquitin pathway in Parkinson's disease. Nature 1998; 395: 451–2.
Polymeropoulos MH, Lavedan C, Leroy E, Ide SE, Dehejia A, Dutra A, et al. Mutation in the alpha-synuclein gene identified in families with Parkinson's disease. Science 1997; 276: 2045–7.
Jenner P . Parkinson's disease, pesticides and mitochondrial dysfunction. Trends Neurosci 2001; 24: 245–7.
Sherer TB, Betarbet R, Greenamyre JT . Environment, mitochondria, and Parkinson's disease. Neuroscientist 2002; 8: 192–7.
Tipton KF, Singer TP . Advances in our understanding of the mechanisms of the neurotoxicity of MPTP and related compounds. J Neurochem 1993; 61: 1191–206.
Schapira AH . Mitochondrial function and neurotoxicity. Curr Opin Neurol 1994; 7: 531–6.
Marey-Semper I, Gelman M, Levi-Strauss M . A selective toxicity toward cultured mesencephalic dopaminergic neurons is induced by the synergistic effects of energetic metabolism impairment and NMDA receptor activation. J Neurosci 1995; 15: 5912–8.
Mizuno Y, Ohta S, Tanaka M, Takamiya S, Suzuki K, Sato T, et al. Deficiencies in complex 1 subunits of the respiratory chain in Parkinson's disease. Biochem Biophys Res Commun 1989; 163: 1450–5.
Heikkila RE, Nicklas WJ, Vyas I, Duvoisin RC . Dopaminergic toxicity of rotenone and the 1-methyl-4-phenylpyridinium ion after their stereotaxic administration to rats: implication for the mechanism of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine toxicity. Neurosci Lett 1985; 62: 389–94.
Ferrante RJ, Schulz JB, Kowall NW, Beal MF . Systemic administration of rotenone produces selective damage in the striatum and globus pallidus, but not in the substantia nigra. Brain Res 1997; 753: 157–62.
Thiffault C, Langston JW, Di Monte DA . Increased striatal dopamine turnover following acute administration of rotenone to mice. Brain Res 2000; 885: 283–8.
Betarbet R, Sherer TB, MacKenzie G, Garcia-Osuna M, Panov AV, Greenamyre JT . Chronic systemic pesticide exposure reproduces features of Parkinson's disease. Nat Neurosci 2000; 3: 1301–6.
Alam M, Schmidt WJ . Rotenone destroys dopaminergic neurons and induces parkinsonian symptoms in rats. Behav Brain Res 2002; 136: 317–24.
Sherer TB, Kim JH, Betarbet R, Greenamyre JT . Subcutaneous rotenone exposure causes highly selective dopaminergic degeneration and alpha-synuclein aggregation. Exp Neurol 2003; 179: 9–16.
Shimura H, Hattori N, Kubo S, Mizuno Y, Asakawa A, Minoshima S, et al. Familial Parkinson disease gene product, parkin, is a ubiquitin-protein ligase. Nat Genet 2000; 25: 302–5.
Shimura H, Schlossmacher MG, Hattori N, Frosch MP, Trockenbacher A, Schneider R, et al. Ubiquitination of a new form of α-synuclein by Parkin from human brain: implications for Parkinson's disease. Science 2001; 293: 263–9.
McNaught KSP, Olanow CW, Halliwell B, Isacson O, Jenner P . Failure of the ubiquitin-proteasome system in Parkinson's disease. Nat Rev Neurosci 2001; 2: 589–94.
Wang RG, Zhu XZ . Subtoxic concentration of manganese synergistically potentiates 1-14 methyl-4-phenylpyridinium-induced neurotoxicity in PC12 cells. Brain Res 2003; 961: 131–8.
Zhou LJ, Zhu XZ . Reactive oxygen species-induced apoptosis in PC12 cells and protective effect of Bilobalide. J Pharmacol Exp Ther 2000; 293: 982–5.
Hirata Y, Adachi K, Kiuchi K . Activation of JNK pathway and induction of apoptosis by manganese in PC12 cells. J Neurochem 1998; 71: 1607–15.
Mefford IN, Giberg M, Barchas JD . Simultaneous determination of catecholamines and unconjugated 3,4-dihydroxyphenylacetic acid (DOPAC) by ion-pairing reverse phase high performance liquid chromatography with electrochemical detection. Anal Biochem 1980; 104: 469–72.
Zhu XZ, Luo LG . Effect of nitroprusside (nitric oxide) on endogenous dopamine release from rat striatal slices. J Neurochem 1992; 59: 932–5.
Wu WR, Zhu XZ, Guan HJ, Wang RG, Ji XQ . Neuroprotective rather than neurorescue or neurorestorative effect of selegiline against MPTP-induced dopaminergic toxicity. Acta Pharmacol Sin 1999; 20: 146–50.
Bal-Price A, Brown GC . Nitric-oxide-induced necrosis and apoptosis in PC12 cells mediated by mitochondria. J Neurochem 2000; 75: 1455–64.
Richter C, Gogvadze V, Laffranchi R, Schlapbach R, Schweizer M, Suter M, et al. Oxidants in mitochondria: from physiology to disease. Biochim Biophys Acta 1995; 1271: 67–74.
Takeshige K, Minakami S . NADH- and NADPH-dependent formation of superoxide anions by bovine heart submitochondrial particles and NADH-ubiquinone reductase preparation. Biochem J 1979; 180: 129–35.
Turrens JF, Boveris A . Generation of superoxide anion by the NADH dehydrogenase of bovine heart mitochondria. Biochem J 1980; 191: 421–7.
Hasegawa E, Takeshige K, Oishi T, Murai Y, Minakami S . 1-Methyl-4-phenylpyridinium (MPP+) induces NADH-dependent superoxide formation and enhances NADH-dependent lipid peroxidation in bovine heart submitochondrial particles. Biochem Biophys Res Commun 1990; 170: 1049–55.
Packer MA, Miesel R, Murphy MP . Exposure to the parkinsonian neurotoxin 1-methyl-4-phenylpyridinium (MPP+) and nitric oxide simultaneously causes cyclosporin A-sensitive mitochondrial calcium efflux and depolarisation. Biochem Pharmacol 1996; 51: 267–73.
Sipos I, Tretter L, Adam-Vizi V . Quantitative relationship between inhibition of respiratory complexes and formation of reactive oxygen species in isolated nerve terminals. J Neurochem 2003; 84: 112–8.
Budd SL, Nicholls DG . Mitochondria, calcium regulation, and acute glutamate excitotoxicity in cultured cerebellar granule cells. J Neurochem 1996; 67: 2282–91.
Rottenberg H, Wu S . Quantitative assay by flow cytometry of the mitochondrial membrane potential in intact cells. Biochim Biophys Acta 1998; 1404: 393–404.
Chinopoulos C, Tretter L, Adam-Vizi V . Depolarization of in situ mitochondria due to hydrogen peroxide-induced oxidative stress in nerve terminals: inhibition of alpha-ketoglutarate dehydrogenase. J Neurochem 1999; 73: 220–8.
Lotharius J, O'Malley KL . The parkinsonism-inducing drug 1-methyl-4-phenylpyridinium triggers intracellular dopamine oxidation. A novel mechanism of toxicity. J Biol Chem 2000; 275: 38581–8.
Brautigam M, Laschinski G, Kittner B, Herken H . Effect of apomorphine, alpha-methylparatyrosine, haloperidol and reserpine on DOPA production in clonal cell lines (PC-12 and N1E-115). Biochem Pharmacol 1985; 34: 941–7.
Fornstedt B . Role of catechol autooxidation in the degeneration of dopamine neurons. Acta Neurol Scand Suppl 1990; 129: 12–4.
Maker HS, Weiss C, Silides DJ, Cohen G . Coupling of dopamine oxidation (monoamine oxidase activity) to glutathione oxidation via the generation of hydrogen peroxide in rat brain homogenates. J Neurochem 1981; 36: 589–93.
Graham DG . Oxidative pathways for catecholamines in the genesis of neuromelanin and cytotoxic quinones. Mol Pharmacol 1978; 14: 633–43.
Kitazawa M, Wagner JR, Kirby ML, Anantharam V, Kanthasamy AG . Oxidative stress and mitochondrial-mediated apoptosis in dopaminergic cells exposed to methylcyclo-pentadienyl manganese tricarbonyl. J Pharmacol Exp Ther 2002; 302: 26–35.
Bywood PT, Johnson SM . Mitochondrial complex inhibitors preferentially damage substantia nigra dopamine neurons in rat brain slices. Exp Neurol 2003; 179: 47–59.
Liu Y, Peter D, Roghani A, Schuldiner S, Prive GG, Eisenberg D, et al. A cDNA that suppresses MPP+ toxicity encodes a vesicular amine transporter. Cell 1992; 70: 539–51.
Author information
Authors and Affiliations
Corresponding author
Additional information
Work supported by a grant from the Chinese Academy of Science and Grant G (1998) 051108 from the Ministry of Sciences and Technology of China.
Rights and permissions
About this article
Cite this article
Liu, Hq., Zhu, Xz. & Weng, Eq. Intracellular dopamine oxidation mediates rotenone-induced apoptosis in PC12 cells. Acta Pharmacol Sin 26, 17–26 (2005). https://doi.org/10.1111/j.1745-7254.2005.00003.x
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1111/j.1745-7254.2005.00003.x
Keywords
This article is cited by
-
Viability of HepG2 and MCF-7 cells is not correlated with mitochondrial bioenergetics
Scientific Reports (2023)
-
Neurotoxicity of pesticides
Acta Neuropathologica (2019)
-
Characteristics of carbonate cementation in clastic rocks from the Chang 6 sandbody of Yanchang Formation, southern Ordos Basin
Science China Earth Sciences (2012)
-
High level interleukin-6 in the medium of human pancreatic cancer cell culture suppresses production of neurotransmitters by PC12 cell line
Metabolic Brain Disease (2012)
-
Dopamine Selectively Sensitizes Dopaminergic Neurons to Rotenone-Induced Apoptosis
Neurochemical Research (2008)