Top

Published in:

01-10-2014 | Neurology and Preclinical Neurological Studies - Original Article

6-Hydroxydopamine impairs mitochondrial function in the rat model of Parkinson’s disease: respirometric, histological, and behavioral analyses

Authors: Andreas Kupsch, Werner Schmidt, Zemfira Gizatullina, Grazyna Debska-Vielhaber, Jürgen Voges, Frank Striggow, Patricia Panther, Herbert Schwegler, Hans-Jochen Heinze, Stefan Vielhaber, Frank Norbert Gellerich

Published in: Journal of Neural Transmission | Issue 10/2014

Abstract

Mitochondrial defects have been shown to be associated with the pathogenesis of Parkinson’s disease (PD). Yet, experience in PD research linking mitochondrial dysfunction, e.g., deregulation of oxidative phosphorylation, with neuronal degeneration and behavioral changes is rather limited. Using the 6-hydroxydopamine (6-OHDA) rat model of PD, we have investigated the potential role of mitochondria in dopaminergic neuronal cell death in the substantia nigra pars compacta by high-resolution respirometry. Mitochondrial function was correlated with the time course of disease-related motor behavior asymmetry and dopaminergic neuronal cell loss, respectively. Unilateral 6-OHDA injections (>2.5 μg/2 μl) into the median forebrain bundle induced an impairment of oxidative phosphorylation due to a decrease in complex I activity. This was indicated by increased flux control coefficient. During the period of days 2–21, a progressive decrease in respiratory control ratio of up to −58 % was observed in the lesioned compared to the non-lesioned substantia nigra of the same animals. This decrease was associated with a marked uncoupling of oxidative phosphorylation. Mitochondrial dysfunction, motor behavior asymmetry, and dopaminergic neuronal cell loss correlated with dosage (1.25–5 μg/2 μl). We conclude that high-resolution respirometry may allow the detection of distinct mitochondrial dysfunction as a suitable surrogate marker for the preclinical assessment of potential neuroprotective strategies in the 6-OHDA model of PD.

Baloh RH, Salavaggione E, Milbrandt J, Pestronk A (2007) Familial Parkinsonism and ophthalmoplegia from a mutation in the mitochondrial DNA helicase twinkle. Arch Neurol 64(7):998–1000. doi:10.1001/archneur.64.7.998 PubMedCrossRef

Banerjee R, Starkov AA, Beal MF, Thomas B (2009) Mitochondrial dysfunction in the limelight of Parkinson’s disease pathogenesis. Biochim Biophys Acta 1792(7):651–663. doi:10.1016/j.bbadis.2008.11.007 PubMedPubMedCentralCrossRef

Bender A, Krishnan KJ, Morris CM, Taylor GA, Reeve AK, Perry RH, Jaros E, Hersheson JS, Betts J, Klopstock T, Taylor RW, Turnbull DM (2006) High levels of mitochondrial DNA deletions in substantia nigra neurons in aging and Parkinson disease. Nat Genet 38(5):515–517. doi:10.1038/ng1769 PubMedCrossRef

Berkich DA, Ola MS, Cole J, Sweatt AJ, Hutson SM, LaNoue KF (2007) Mitochondrial transport proteins of the brain. J Neurosci Res 85(15):3367–3377. doi:10.1002/jnr.21500 PubMedCrossRef

Bové J, Perier C (2012) Neurotoxin-based models of Parkinson’s disease. Neuroscience 211:51–76. doi:10.1016/j.neuroscience.2011.10.057 PubMedCrossRef

Brand MD, Nicholls DG (2011) Assessing mitochondrial dysfunction in cells. Biochem J 435(2):297–312. doi:10.1042/BJ20110162 PubMedPubMedCentralCrossRef

Bredesen DE, Rao RV, Mehlen P (2006) Cell death in the nervous system. Nature 443(7113):796–802. doi:10.1038/nature05293 PubMedPubMedCentralCrossRef

Chaudhuri KR, Healy DG, Schapira AH; National Institute for Clinical E (2006) Non-motor symptoms of Parkinson’s disease: diagnosis and management. Lancet Neurol 5 (3):235–245. doi:10.1016/S1474-4422(06)70373-8

Cohen G (1984) Oxy-radical toxicity in catecholamine neurons. Neurotoxicology 5(1):77–82PubMed

de Lau LM, Giesbergen PC, de Rijk MC, Hofman A, Koudstaal PJ, Breteler MM (2004) Incidence of parkinsonism and Parkinson disease in a general population: the Rotterdam Study. Neurology 63(7):1240–1244PubMedCrossRef

dos Santos El-Bachá R, Daval J-L, Koziel V, Netter P, Minn A (2001) Toxic effects of apomorphine on rat cultured neurons and glial C6 cells, and protection with antioxidants. Biochem Pharmacol 61(1):73-85. doi:10.1016/S0006-2952(00)00524-4

Ekstrand MI, Terzioglu M, Galter D, Zhu S, Hofstetter C, Lindqvist E, Thams S, Bergstrand A, Hansson FS, Trifunovic A, Hoffer B, Cullheim S, Mohammed AH, Olson L, Larsson NG (2007) Progressive parkinsonism in mice with respiratory-chain-deficient dopamine neurons. Proc Natl Acad Sci USA 104(4):1325–1330. doi:10.1073/pnas.0605208103 PubMedPubMedCentralCrossRef

Fahn S (2003) Description of Parkinson’s disease as a clinical syndrome. Ann N Y Acad Sci 991:1–14PubMedCrossRef

Gassen M, Glinka Y, Pinchasi B, Youdim MBH (1996) Apomorphine is a highly potent free radical scavenger in rat brain mitochondrial fraction. Eur J Pharmacol 308(2):219–225. doi:10.1016/0014-2999(96)00291-9 PubMedCrossRef

Gellerich FN, Kunz WS, Bohnensack R (1990) Estimation of flux control coefficients from inhibitor titrations by non-linear regression. FEBS Lett 274(1–2):167–170PubMedCrossRef

Gellerich FN, Deschauer M, Chen Y, Müller T, Neudecker S, Zierz S (2002) Mitochondrial respiratory rates and activities of respiratory chain complexes correlate linearly with heteroplasmy of deleted mtDNA without threshold and independently of deletion size. Biochim Biophys Acta 1556(1):41–52PubMedCrossRef

Gellerich FN, Gizatullina Z, Trumbekaite S, Korzeniewski B, Gaynutdinov T, Seppet E, Vielhaber S, Heinze HJ, Striggow F (2012) Cytosolic Ca²⁺ regulates the energization of isolated brain mitochondria by formation of pyruvate through the malate-aspartate shuttle. Biochem J 443(3):747–755. doi:10.1042/BJ20110765 PubMedCrossRef

Gellerich FN, Gizatullina Z, Gainutdinov T, Muth K, Seppet E, Orynbayeva Z, Vielhaber S (2013) The control of brain mitochondrial energization by cytosolic calcium: the mitochondrial gas pedal. IUBMB Life 65(3):180–190. doi:10.1002/iub.1131 PubMedCrossRef

Giordano S, Lee J, Darley-Usmar VM, Zhang J (2012) Distinct effects of rotenone, 1-methyl-4-phenylpyridinium and 6-hydroxydopamine on cellular bioenergetics and cell death. PLoS ONE 7(9):e44610. doi:10.1371/journal.pone.0044610 PubMedPubMedCentralCrossRef

Glinka YY, Youdim MB (1995) Inhibition of mitochondrial complexes I and IV by 6-hydroxydopamine. Eur J Pharmacol 292(3–4):329–332PubMed

Glinka Y, Tipton KF, Youdim MB (1998) Mechanism of inhibition of mitochondrial respiratory complex I by 6-hydroxydopamine and its prevention by desferrioxamine. Eur J Pharmacol 351(1):121–129PubMedCrossRef

Gnaiger E (2001) Bioenergetics at low oxygen: dependence of respiration and phosphorylation on oxygen and adenosine diphosphate supply. Respir Physiol 128(3):277–297PubMedCrossRef

Heinrich R, Rapoport TA (1974) A linear steady-state treatment of enzymatic chains. Critique of the crossover theorem and a general procedure to identify interaction sites with an effector. Eur J Biochem/FEBS 42(1):97–105CrossRef

Hökfelt T, Ungerstedt U (1973) Specificity of 6-hydroxydopamine induced degeneration of central monoamine neurones: an electron and fluorescence microscopic study with special reference to intracerebral injection on the nigro-striatal dopamine system. Brain Res 60(2):269–297PubMedCrossRef

Kacser H, Burns JA (1973) The control of flux. Symp Soc Exp Biol 27:65–104PubMed

Khaliulin I, Schneider A, Houminer E, Borman JB, Schwalb H (2004) Apomorphine prevents myocardial ischemia/reperfusion-induced oxidative stress in the rat heart. Free Radic Biol Med 37(7):969–976. doi:10.1016/j.freeradbiomed.2004.06.029 PubMedCrossRef

Koh H, Chung J (2012) PINK1 as a molecular checkpoint in the maintenance of mitochondrial function and integrity. Mol Cells 34(1):7–13. doi:10.1007/s10059-012-0100-8 PubMedPubMedCentralCrossRef

Kraytsberg Y, Kudryavtseva E, McKee AC, Geula C, Kowall NW, Khrapko K (2006) Mitochondrial DNA deletions are abundant and cause functional impairment in aged human substantia nigra neurons. Nat Genet 38(5):518–520. doi:10.1038/ng1778 PubMedCrossRef

Kunz WS, Kuznetsov AV, Clark JF, Tracey I, Elger CE (1999) Metabolic consequences of the cytochrome c oxidase deficiency in brain of copper-deficient Mo(vbr) mice. J Neurochem 72(4):1580–1585PubMedCrossRef

Kupsch A, Loschmann PA, Sauer H, Arnold G, Renner P, Pufal D, Burg M, Wachtel H, ten Bruggencate G, Oertel WH (1992) Do NMDA receptor antagonists protect against MPTP-toxicity? Biochemical and immunocytochemical analyses in black mice. Brain Res 592(1–2):74–83PubMedCrossRef

Kuznetsov AV, Tiivel T, Sikk P, Kaambre T, Kay L, Daneshrad Z, Rossi A, Kadaja L, Peet N, Seppet E, Saks VA (1996) Striking differences between the kinetics of regulation of respiration by ADP in slow-twitch and fast-twitch muscles in vivo. Eur J Biochem/FEBS 241(3):909–915CrossRef

Kuznetsov AV, Veksler V, Gellerich FN, Saks V, Margreiter R, Kunz WS (2008) Analysis of mitochondrial function in situ in permeabilized muscle fibers, tissues and cells. Nat Protoc 3(6):965–976. doi:10.1038/nprot.2008.61 PubMedCrossRef

Kyriazis M (2003) Neuroprotective, anti-apoptotic effects of apomorphine. J Anti Aging Med 6(1):21–28. doi:10.1089/109454503765361551 PubMedCrossRef

Lemasters JJ, Nieminen AL (1997) Mitochondrial oxygen radical formation during reductive and oxidative stress to intact hepatocytes. Biosci Rep 17(3):281–291PubMedCrossRef

Lin MT, Cantuti-Castelvetri I, Zheng K, Jackson KE, Tan YB, Arzberger T, Lees AJ, Betensky RA, Beal MF, Simon DK (2012) Somatic mitochondrial DNA mutations in early Parkinson and incidental Lewy body disease. Ann Neurol 71(6):850–854. doi:10.1002/ana.23568 PubMedPubMedCentralCrossRef

Luoma P, Melberg A, Rinne JO, Kaukonen JA, Nupponen NN, Chalmers RM, Oldfors A, Rautakorpi I, Peltonen L, Majamaa K, Somer H, Suomalainen A (2004) Parkinsonism, premature menopause, and mitochondrial DNA polymerase gamma mutations: clinical and molecular genetic study. Lancet 364(9437):875–882. doi:10.1016/S0140-6736(04)16983-3 PubMedCrossRef

Miller RL, James-Kracke M, Sun GY, Sun AY (2009) Oxidative and inflammatory pathways in Parkinson’s disease. Neurochem Res 34(1):55–65. doi:10.1007/s11064-008-9656-2 PubMedCrossRef

Musatov A, Robinson NC (2012) Susceptibility of mitochondrial electron-transport complexes to oxidative damage. Focus on cytochrome c oxidase. Free Radic Res 46(11):1313–1326. doi:10.3109/10715762.2012.717273 PubMedCrossRef

Olszewska A, Szewczyk A (2013) Mitochondria as a pharmacological target: magnum overview. IUBMB Life 65(3):273–281. doi:10.1002/iub.1147 PubMedCrossRef

Paxinos G, Watson C (2007) The rat brain in stereotaxic coordinates. Academic, New York

Schapira AH (2008) Mitochondria in the aetiology and pathogenesis of Parkinson’s disease. Lancet Neurol 7(1):97–109. doi:10.1016/S1474-4422(07)70327-7 PubMedCrossRef

Schapira AH, Cooper JM, Dexter D, Jenner P, Clark JB, Marsden CD (1989) Mitochondrial complex I deficiency in Parkinson’s disease. Lancet 1(8649):1269PubMedCrossRef

Seppet E, Gizatullina Z, Trumbeckaite S, Zierz S, Striggow F, Gellerich FN (2007) Mitochondrial medicine: the central role of cellular energetic depression and mitochondria in cell pathophysiology. In: Molecular System Bioenergetics. Wiley, New York, pp 479–520. doi:10.1002/9783527621095.ch15

Soong NW, Hinton DR, Cortopassi G, Arnheim N (1992) Mosaicism for a specific somatic mitochondrial DNA mutation in adult human brain. Nat Genet 2(4):318–323. doi:10.1038/ng1292-318 PubMedCrossRef

Tobon-Velasco JC, Limon-Pacheco JH, Orozco-Ibarra M, Macias-Silva M, Vazquez-Victorio G, Cuevas E, Ali SF, Cuadrado A, Pedraza-Chaverri J, Santamaria A (2013a) 6-OHDA-induced apoptosis and mitochondrial dysfunction are mediated by early modulation of intracellular signals and interaction of Nrf2 and NF-kappaB factors. Toxicology 304:109–119. doi:10.1016/j.tox.2012.12.011 PubMedCrossRef

Tobon-Velasco JC, Palafox-Sanchez V, Mendieta L, Garcia E, Santamaria A, Chamorro-Cevallos G, Limon ID (2013b) Antioxidant effect of Spirulina (Arthrospira) maxima in a neurotoxic model caused by 6-OHDA in the rat striatum. J Neural Transm. doi:10.1007/s00702-013-0976-2 PubMed

Trumbeckaite S, Gizatullina Z, Arandarcikaite O, Rohnert P, Vielhaber S, Malesevic M, Fischer G, Seppet E, Striggow F, Gellerich FN (2012) Oxygen glucose deprivation causes mitochondrial dysfunction in cultivated rat hippocampal slices: protective effects of CsA, its immunosuppressive congener [D-Ser](8)CsA, the novel non-immunosuppressive cyclosporin derivative Cs9, and the NMDA receptor antagonist MK 801. Mitochondrion. doi:10.1016/j.mito.2012.07.110 PubMed

Ungerstedt U (1968) 6-Hydroxy-dopamine induced degeneration of central monoamine neurons. Eur J Pharmacol 5(1):107–110PubMedCrossRef

Ungerstedt U, Arbuthnott GW (1970) Quantitative recording of rotational behavior in rats after 6-hydroxy-dopamine lesions of the nigrostriatal dopamine system. Brain Res 24(3):485–493PubMedCrossRef

Vlamings R, Visser-Vandewalle V, Kozan R, Kaplan S, Steinbusch HW, Temel Y (2009) Bilateral high frequency stimulation of the subthalamic nucleus normalizes COX activity in the substantia nigra of Parkinsonian rats. Brain Res 1288:143–148. doi:10.1016/j.brainres.2009.06.091 PubMedCrossRef

Webster KA (2012) Mitochondrial membrane permeabilization and cell death during myocardial infarction: roles of calcium and reactive oxygen species. Futur Cardiol 8(6):863–884. doi:10.2217/fca.12.58 CrossRef

Wiedemann FR, Winkler K, Kuznetsov AV, Bartels C, Vielhaber S, Feistner H, Kunz WS (1998) Impairment of mitochondrial function in skeletal muscle of patients with amyotrophic lateral sclerosis. J Neurol Sci 156(1):65–72PubMedCrossRef

Title: 6-Hydroxydopamine impairs mitochondrial function in the rat model of Parkinson’s disease: respirometric, histological, and behavioral analyses
Authors: Andreas Kupsch
Werner Schmidt
Zemfira Gizatullina
Grazyna Debska-Vielhaber
Jürgen Voges
Frank Striggow
Patricia Panther
Herbert Schwegler
Hans-Jochen Heinze
Stefan Vielhaber
Frank Norbert Gellerich
Publication date: 01-10-2014
Publisher: Springer Vienna
Published in: Journal of Neural Transmission / Issue 10/2014
Print ISSN: 0300-9564
Electronic ISSN: 1435-1463
DOI: https://doi.org/10.1007/s00702-014-1185-3

Keynote webinar | Spotlight on medication adherence

Springer Medicine

6-Hydroxydopamine impairs mitochondrial function in the rat model of Parkinson’s disease: respirometric, histological, and behavioral analyses

Abstract

Keynote webinar | Spotlight on medication adherence

Springer Medicine

Abstract

Please log in to get access to this content

Other articles of this Issue 10/2014

Subacute peripheral neuropathy under duodopa therapy without cobalamin deficiency and despite supplementation

Automation of o-dianisidine assay for ceruloplasmin activity analyses: usefulness of investigation in Wilson’s disease and in hepatic encephalopathy

Carotid artery thickening and neurocirculatory abnormalities in de novo Parkinson disease

Subthalamic nucleus involvement in executive functions with increased cognitive load: a subthalamic nucleus and anterior cingulate cortex depth recording study

Salubrinal, ER stress inhibitor, attenuates kainic acid-induced hippocampal cell death

Physiological effects of cigarette smoking in the limbic system revealed by 3 tesla magnetic resonance spectroscopy