Abstract
Heterogenous diseases such as Parkinson’s disease (PD) needs an efficient animal model to enhance understanding of the underlying mechanisms and to develop therapeutics. MPTP (1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine), a neurotoxin, has been widely used to replicate the pathophysiology of PD in rodents, however, the knowledge about its effects on energy metabolism is limited. Moreover, susceptibility to different dose regimens of MPTP also varies among mice strains. Thus, the present study compares the effect of acute and sub-acute MPTP administration on mitochondrial functions in C57BL/6 and BALB/c mice. In addition, activity of enzymes involved in energy metabolism was also studied along with behavioural alterations. The findings show that acute dose of MPTP in C57BL/6 mice had more profound effect on the activity of electron transport chain complexes. Further, the activity of MAO-B was increased following acute and sub-acute MPTP administration in C57BL/6 mice. However, no significant change was observed in BALB/c mice. Acute MPTP treatment resulted in decreased mitochondrial membrane potential along with increased swelling of mitochondria in C57BL/6 mice. In addition, perturbations were observed in hexokinase, the rate limiting enzyme of glycolysis and pyruvate dehydrogenase, the enzymes that connects glycolysis and TCA cycle. The activity of TCA cycle enzymes; citrate synthase, aconitase, isocitrate dehydrogenase and fumarase were also altered following MPTP intoxication. Furthermore, acute MPTP administration led to drastic reduction in dopamine levels in striatum of C57BL/6 as compared to BALB/c mice. Behavioral tests such as open field, narrow beam walk and footprint analysis revealed severe impairment in locomotor activity in C57BL/6 mice. These results clearly demonstrate that C57BL/6 strain is more vulnerable to MPTP-induced mitochondrial dysfunctions, perturbations in energy metabolism and motor defects as compared to BALB/c strain. Thus, the findings suggest that the dose and strain of mice need to be considered for pre-clinical studies using MPTP-induced model of Parkinson’s disease.
Graphical abstract
Highlights
-
MPTP-induced mitochondrial perturbations were studied in C57BL/6 and BALB/c mice.
-
C57BL/6 mice had higher susceptibility to MPTP in terms of mitochondrial dysfunctions and motor defects.
-
Motor impairments were accompanied by decrease in dopamine levels in C57BL/6 mice following acute MPTP administration.
-
Acute and sub-acute MPTP administration impaired glycolytic and TCA cycle enzymes.
-
The findings suggest acute MPTP administration in C57BL/6 mice is a suitable toxin-induced PD model for pre-clinical studies.
Similar content being viewed by others
Data availability
The data that support the findings of this study are available on reasonable request from the corresponding author.
Abbreviations
- BSA:
-
Bovine serum albumin
- CS:
-
Citrate synthase
- DA:
-
Dopamine
- DMEM:
-
Dulbecco’s Modified Eagle’s Medium
- ETC:
-
Electron transport chain
- EDTA:
-
Ethylene diamine tetra-acetic acid
- HEPES:
-
4-(2-Hydroxyethyl)- 1-piperazineethanesulfonic acid
- MAO:
-
Monoamine oxidase
- MPTP:
-
1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine
- PD:
-
Parkinson’s Disease
- PDHc:
-
Pyruvate dehydrogenase complex
- TCA:
-
Tri-carboxylic acid
- SN:
-
Substantia nigra
References
Ahmed SS, Santosh W, Kumar S, Christlet HTT (2009) Metabolic profiling of Parkinson’s disease: evidence of biomarker from gene expression analysis and rapid neural network detection. J Biomed Sci 16:63
Anandhan A, Jacome MS, Lei S, Hernandez-Franco P, Pappa A, Panayiotidis MI, Powers R, Franco R (2017) Metabolic dysfunction in Parkinson’s Disease: Bioenergetics, redox homeostasis and central carbon metabolism. Brain Res Bull 133:12–30
Berman SB, Hastings TG (2001) Dopamine oxidation alters mitochondrial respiration and induces permeability transition in brain mitochondria. J Neurochem 73:1127–1137
Blin O, Desnuelle C, Rascol O, Borg M, Paul HPS, Azulay JP, Billé F, Figarella D, Coulom F, Pellissier JF, Montastruc JL, Chatel M, Serratrice G (1994) Mitochondrial respiratory failure in skeletal muscle from patients with Parkinson’s disease and multiple system atrophy. J Neurol Sci 125:95–101
Braak H, Del Tredici K (2008) Nervous system pathology in sporadic Parkinson disease. Neurology 70:1916–1925
Cenci MA, Whishaw IQ, Schallert T (2002) Animal models of neurological deficits: How relevant is the rat? Nat Rev Neurosci 3:574–579
Church WH (2005) Column chromatography analysis of brain tissue: An advanced laboratory exercise for neuroscience majors. J Undergrad Neurosci Educ 3:A36-41
Cirillo G, Maggio N, Bianco MR, Vollono C, Sellitti S, Papa M (2010) Discriminative behavioral assessment unveils remarkable reactive astrocytosis and early molecular correlates in basal ganglia of 3-nitropropionic acid subchronic treated rats. Neurochem Int 56:152–160
Darios F, Corti O, Lücking CB, Hampe C, Muriel M-P, Abbas N, Gu W-J, Hirsch EC, Rooney T, Ruberg M, Brice A (2003) Parkin prevents mitochondrial swelling and cytochrome c release in mitochondria-dependent cell death. Hum Mol Genet 12:517–526
Davey GP, Peuchen S, Clark JB (1998) Energy thresholds in brain mitochondria: Potential involvement in neurodegeneration. J Biol Chem 273:12753–12757
Eberling JL, Richardson BC, Reed BR, Wolfe N, Jagust WJ (1994) Cortical glucose metabolism in Parkinson’s disease without dementia. Neurobiol Aging 15:329–335
Ellis G, Goldberg DM (1971) An improved manual and semi-automatic assay for NADP-dependent isocitrate dehydrogenase activity, with a description of some kinetic properties of human liver and serum enzyme. Clin Biochem 4:175–185
Fischer DA, Ferger B, Kuschinsky K (2002) Discrimination of morphine- and haloperidol-induced muscular rigidity and akinesia/catalepsy in simple tests in rats. Behav Brain Res 134:317–321
Fowler CJ, Wiberg A, Oreland L, Marcusson J, Winblad B (1980) The effect of age on the activity and molecular properties of human brain monoamine oxidase. J Neural Transm 49:1–20
Gibrat C, Saint-Pierre M, Bousquet M, Lévesque D, Rouillard C, Cicchetti F (2009) Differences between subacute and chronic MPTP mice models: investigation of dopaminergic neuronal degeneration and α-synuclein inclusions. J Neurochem 109:1469–1482
Griffiths DE, Houghton RL (1974) Studies on energy-linked reactions: modified mitochondrial ATPase of oligomycin-resistant mutants of saccharomyces cerevisiae. Eur J Biochem 46:157–167
Gupta D, Kurhe Y, Radhakrishnan M (2014) Antidepressant effects of insulin in streptozotocin induced diabetic mice: Modulation of brain serotonin system. Physiol Behav 129:73–78
Hashimoto M, Rockenstein E, Masliah E (2003) Transgenic models of alpha-synuclein pathology: past, present, and future. Ann N Y Acad Sci 991:171–188
Hauser DN, Mamais A, Conti MM, Primiani CT, Kumaran R, Dillman AA, Langston RG, Beilina A, Garcia JH, Diaz-Ruiz A, Bernier M, Fiesel FC, Hou X, Springer W, Li Y, De Cabo R, Cookson MR (2017) Hexokinases link DJ-1 to the PINK1/parkin pathway. Mol Neurodegener 12:1–17
Ito T, Suzuki K, Uchida K, Nakayama H (2013a) Different susceptibility to 1-methyl-4-phenylpyridium (MPP+)-induced nigro-striatal dopaminergic cell loss between C57BL/6 and BALB/c mice is not related to the difference of monoamine oxidase-B (MAO-B). Exp Toxicol Pathol 65:153–158
Ito T, Uchida K, Nakayama H (2013b) Neuronal or inducible nitric oxide synthase (NOS) expression level is not involved in the different susceptibility to nigro-striatal dopaminergic neurotoxicity induced by 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) between C57BL/6 and BALB/c mice. Exp Toxicol Pathol 65:121–125
Jogdand PS, Singh SK, Christiansen M, Dziegiel MH, Singh S, Theisen M (2012) Flow cytometric readout based on Mitotracker Red CMXRos staining of live asexual blood stage. Malar J 11:235
Kanarek L, Hill RL (1964) The preparation and characterization of fumarase from swine heart and muscle. J Biol Chem 239:4202–4206
Ke CJ, He YH, He HW, Yang X, Li R, Yuan J (2014) A new spectrophotometric assay for measuring pyruvate dehydrogenase complex activity: A comparative evaluation. Anal Methods 6:6381–6388
King TE, Howard RL (1967) Preparations and properties of soluble NADH dehydrogenases from cardiac muscle. Methods Enzymol 10:275–294
King TE, Ohnishi T, Winter DB, Wu JT (1976) Biochemical and EPR probes for structure-function studies of iron sulfur centers of succinate dehydrogenase. Adv Exp Med Biol 74:182–227
Kristián T, Gertsch J, Bates TE, Siesjö BK (2000) Characteristics of the calcium-triggered mitochondrial permeability transition in nonsynaptic brain mitochondria: Effect of cyclosporin A and ubiquinone O. J Neurochem 74:1999–2009
Lee KS, Lee JK, Kim HG, Kim HR (2013) Differential effects of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine on motor behavior and dopamine levels at brain regions in three different mouse strains. Korean J Physiol Pharmacol 17:89–97
Liang LP, Patel M (2004) Iron-sulfur enzyme mediated mitochondrial superoxide toxicity in experimental Parkinson’s disease. J Neurochem 90:1076–1084
Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the Folin phenol reagent. J Biol Chem 193:265–275
Masliah E, Rockenstein E, Veinbergs I, Mallory M, Hashimoto M, Takeda A, Sagara Y, Sisk A, Mucke L (2000) Dopaminergic loss and inclusion body formation in α-synuclein mice: Implications for neurodegenerative disorders. Science 287:1265–1269
Mattson MP, Pedersen WA, Duan W, Culmsee C, Camandola S (1999) Cellular and molecular mechanisms underlying perturbed energy metabolism and neuronal degeneration in Alzheimer’s and Parkinson’s diseases. Ann N Y Acad Sci 893:154–175
McCormack AL, Thiruchelvam M, Manning-Bog AB, Thiffault C, Langston JW, Cory-Slechta DA, Di Monte DA (2002) Environmental risk factors and Parkinson’s disease: Selective degeneration of nigral dopaminergic neurons caused by the herbicide paraquat. Neurobiol Dis 10:119–127
McCoy MK, Kaganovich A, Rudenko IN, Ding J, Cookson MR (2014) Hexokinase activity is required for recruitment of parkin to depolarized mitochondria. Hum Mol Genet 23:145–156
Meredith GE, Rademacher DJ (2011) MPTP mouse models of Parkinson’s disease: An update. J Parkinsons Dis 1:19–33
Mizuno Y, Ohta S, Tanaka M, Takamiya S, Suzuki K, Sato T, Oya H, Ozawa T, Kagawa Y (1989) Deficiencies in Complex I subunits of the respiratory chain in Parkinson’s disease. Biochem Biophys Res Commun 163:1450–1455
Morton RL, Iklé D, White CW (1998) Loss of lung mitochondrial aconitase activity due to hyperoxia in bronchopulmonary dysplasia in primates. Am J Physiol Cell Mol Physiol 274:L127–L133
Orozco JL, Valderrama-Chaparro JA, Pinilla-Monsalve GD, Molina-Echeverry MI, Castaño AMP, Ariza-Araújo Y, Prada SI, Takeuchi Y (2020) Parkinson’s disease prevalence, age distribution and staging in Colombia. Neurol Int 12:9–14
Parker WD, Parks JK, Swerdlow RH (2008) Complex I deficiency in Parkinson’s disease frontal cortex. Brain Res 1189:215–218
Parnetti L, Gaiti A, Polidori MC, Brunetti M, Palumbo B, Chionne F, Cadini D, Cecchetti R, Senin U (1995) Increased cerebrospinal fluid pyruvate levels in Alzheimer’s disease. Neurosci Lett 199:231–233
Perier C, Bové J, Vila M, Przedborski S (2003) The rotenone model of Parkinson’s disease. Trends Neurosci 26:345–346
Przedborski S, Jackson-Lewis V, Naini AB, Jakowec M, Petzinger G, Miller R, Akram M (2001) The parkinsonian toxin 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP): A technical review of its utility and safety. J Neurochem 76:1265–1274
Puka-Sundvall M, Wallin C, Gilland E, Hallin U, Wang X, Sandberg M, Karlsson JO, Blomgren K, Hagberg H (2000) Impairment of mitochondrial respiration after cerebral hypoxia-ischemia in immature rats: Relationship to activation of caspase-3 and neuronal injury. Dev Brain Res 125:43–50
Roberts DJ, Miyamoto S (2015) Hexokinase II integrates energy metabolism and cellular protection: Akting on mitochondria and TORCing to autophagy. Cell Death Differ 22:248–257
Saura DJ, Richards JG, Mahy N (1994) Age-related changes on MAO in Bl/C57 mouse tissues: A quantitative radioautographic study. J Neural Transm Suppl 41:89–94
Schapira AHV (2007) Mitochondrial dysfunction in Parkinson’s disease. Cell Death Differ 14:1261–1266
Schapira AHV, Cooper JM, Dexter D, Clark JB, Jenner P, Marsden CD (1990) Mitochondrial Complex I Deficiency in Parkinson’s Disease. J Neurochem 54:823–827
Sedelis M, Hofele K, Auburger GW, Morgan S, Huston JP, Schwarting RKW (2000) MPTP susceptibility in the mouse: Behavioral, neurochemical, and histological analysis of gender and strain differences. Behav Genet 30:171–182
Seibenhener ML, Wooten MC (2015) Use of the open field maze to measure locomotor and anxiety-like behavior in mice. J vis Exp 6:52434
Sottocasa GL, Kuylenstierna B, Ernster L, Bergstrand A (1967) An electron-transport system associated with the outer membrane of liver mitochondria. A biochemical and morphological study. J Cell Biol 32:415–438
Southwell AL, Ko J, Patterson PH (2009) Intrabody gene therapy ameliorates motor, cognitive, and neuropathological symptoms in multiple mouse models of Huntington’s disease. J Neurosci 29:13589–13602
Spinazzi M, Casarin A, Pertegato V, Salviati L, Angelini C (2012) Assessment of mitochondrial respiratory chain enzymatic activities on tissues and cultured cells. Nat Protoc 7:1235–1246
Bergmeyer HU, Bernt E, Schmidt F, and Stork H (1974). Determination with hexokinase and glucose-6-phosphate dehydrogenase. In 'Methods of Enzymatic Analysis'. 2nd English Edn. (Ed. H. U. Bergmeyer.) pp. 1196–201
Acknowledgements
The financial assistance provided by the University Grants Commission (UGC), New Delhi under the Basic Science Research (UGC Ref. No. F.25-1/2014-15(BSR)/7-209/2009[BSR]) is acknowledged. The authors also acknowledge Department of Biotechnology (DBT), Government of India for the financial support (BT/PR17127/NER/95/453/2015).
Author information
Authors and Affiliations
Contributions
AP and RS conceptualized and designed the study. AP and PG conducted experiments and analyzed the data. All authors contributed in drafting and editing the manuscript.
Corresponding author
Ethics declarations
Conflict of interest
The authors declare no competing conflict of interest.
Additional information
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Rights and permissions
About this article
Cite this article
Pathania, A., Garg, P. & Sandhir, R. Impaired mitochondrial functions and energy metabolism in MPTP-induced Parkinson’s disease: comparison of mice strains and dose regimens. Metab Brain Dis 36, 2343–2357 (2021). https://doi.org/10.1007/s11011-021-00840-2
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11011-021-00840-2