Iranian Journal of Neurology 2015. 14(4):195-203.

Effects of L-arginine pre-treatment in 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine-induced Parkinson’s diseases in Balb/c mice
Javad Hami, Mehran Hosseini, Sekineh Shahi, Nassim Lotfi, Abolfazl Talebi, Mohammad Afshar


Background: Parkinson’s disease (PD) is a common neurodegenerative disease resulting from the degeneration of dopaminergic (DA) neurons in the substantia nigra pars compacta (SNc). Increasing evidence demonstrated that mice treated intranasally with 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) suffered impairments in motor functions associated with disruption of DA neurons in SNc conceivably analogous to those observed in PD. L-arginine has been proposed as a novel neuroprotective agent that plays protective roles in several models of neuronal cellular damage. This study aimed to evaluate the effects of L-arginine on the numerical density of dark neurons (DNs) in the SNc of Balb/c mice subjected to MPTP administration.
Methods: In the present study, we demonstrated that repeated treatment with L-arginine (300 mg/kg, i.p.) during 7 consecutive days attenuated the production of DNs in SNc of adult male Balb/c mice infused with a single intranasal administration of MPTP (1 mg/nostril).
Results: Pre-treatment with L-arginine significantly decreased the numerical density of DNs in SNc of mice 21 days after intranasal MPTP administration.
Conclusion: This investigation provides new insights in experimental models of PD, indicating that L-arginine represents a potential neuroprotective agent for the prevention of DA neuron degeneration in SNc observed in PD patients.


Parkinson Disease;1-Methyl-4-phenyl-1-2- 6-tetrahydropyridine; BALB C Mice; Protective Agents

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Thenganatt MA, Jankovic J. Parkinson disease subtypes. JAMA Neurol 2014;71(4): 499-504.

Alves G, Forsaa EB, Pedersen KF, Dreetz GM, Larsen JP. Epidemiology of Parkinson's disease. J Neurol 2008; 255(Suppl 5): 18-32.

Schapira AH, Jenner P. Etiology and pathogenesis of Parkinson's disease. Mov Disord 2011; 26(6): 1049-55.

Mandel S, Grunblatt E, Riederer P, Gerlach M, Levites Y, Youdim MB. Neuroprotective strategies in Parkinson's disease : an update on progress. CNS Drugs 2003; 17(10): 729-62.

Jenner P. Oxidative stress as a cause of Parkinson's disease. Acta Neurol Scand Suppl 1991; 136: 6-15.

Roy A, Ghosh A, Jana A, Liu X, Brahmachari S, Gendelman HE, et al. Sodium phenylbutyrate controls neuroinflammatory and antioxidant activities and protects dopaminergic neurons in mouse models of Parkinson's disease. PLoS One 2012; 7(6): e38113.

Barnett EM, Perlman S. The olfactory nerve and not the trigeminal nerve is the major site of CNS entry for mouse hepatitis virus, strain JHM. Virology 1993; 194(1): 185-91.

Hastings L, Evans JE. Olfactory primary neurons as a route of entry for toxic agents into the CNS. Neurotoxicology 1991; 12(4):707-14.

Zatta P, Favarato M, Nicolini M. Deposition of aluminum in brain tissues of rats exposed to inhalation of aluminum acetylacetonate.Neuroreport 1993; 4(9): 1119-22.

Tjalve H, Henriksson J, Tallkvist J, Larsson BS, Lindquist NG. Uptake of manganese and cadmium from the nasal mucosa into the central nervous system via olfactory pathways in rats. Pharmacol Toxicol 1996; 79(6): 347-56.

Doty RL. The olfactory vector hypothesis of neurodegenerative disease: isit viable? Ann Neurol 2008; 63(1): 7-15.

Doty RL, Deems DA, Stellar S. Olfactory dysfunction in parkinsonism: a general deficit unrelated to neurologic signs, disease stage, or disease duration. Neurology 1988;38(8): 1237-44.

Braak H, Ghebremedhin E, Rub U, Bratzke H, Del TK. Stages in the development of Parkinson's disease-related pathology. Cell Tissue Res 2004; 318(1): 121-34.

Li LH, Qin HZ, Wang JL, Wang J, Wang XL, Gao GD. Axonal degeneration of nigra- striatum dopaminergic neurons induced by 1-methyl-4-phenyl-1,2,3,6- tetrahydropyridine in mice. J Int Med Res 2009; 37(2): 455-63.

He XJ, Nakayama H, Dong M, Yamauchi H, Ueno M, Uetsuka K, et al. Evidence of apoptosis in the subventricular zone and rostral migratory stream in the MPTP mouse model of Parkinson disease. J Neuropathol Exp Neurol 2006; 65(9): 873-82.

Prediger RD, Aguiar AS, Jr., Rojas- Mayorquin AE, Figueiredo CP, Matheus FC, Ginestet L, et al. Single intranasal administration of 1-methyl-4-phenyl- 1,2,3,6-tetrahydropyridine in C57BL/6 mice models early preclinical phase of Parkinson's disease. Neurotox Res 2010; 17(2): 114-29.

Dluzen DE, Kefalas G. The effects of intranasal infusion of 1-methyl-4-phenyl- 1,2,3,6-tetrahydropyridine (MPTP) upon catecholamine concentrations within olfactory bulbs and corpus striatum of male mice. Brain Res 1996; 741(1-2): 215-9.

Prediger RD, Aguiar AS, Jr., Moreira EL, Matheus FC, Castro AA, Walz R, et al. The intranasal administration of 1-methyl-4- phenyl-1,2,3,6-tetrahydropyridine (MPTP): a new rodent model to test palliative and neuroprotective agents for Parkinson's disease. Curr Pharm Des 2011; 17(5): 489-507.

Prediger RD, Batista LC, Medeiros R, Pandolfo P, Florio JC, Takahashi RN. The risk is in the air: Intranasal administration of MPTP to rats reproducing clinical features of Parkinson's disease. Exp Neurol 2006;202(2): 391-403.

Prediger RD, Rial D, Medeiros R, Figueiredo CP, Doty RL, Takahashi RN. Risk is in the air: an intranasal MPTP (1- methyl-4-phenyl-1,2,3,6-tetrahydropyridine) rat model of Parkinson's disease. Ann N Y Acad Sci 2009; 1170: 629-36.

Ransom BR, Kunis DM, Irwin I, Langston JW. Astrocytes convert the parkinsonism inducing neurotoxin, MPTP, to its active metabolite, MPP+. Neurosci Lett 1987; 75(3): 323-8.

Cui M, Aras R, Christian WV, Rappold PM, Hatwar M, Panza J, et al. The organic cation transporter-3 is a pivotal modulator of neurodegeneration in the nigrostriatal dopaminergic pathway. Proc Natl Acad Sci U S A 2009; 106(19): 8043-8.

Kopin IJ. Features of the dopaminergic neurotoxin MPTP. Ann N Y Acad Sci 1992; 648: 96-104.

Jackson-Lewis V, Jakowec M, Burke RE, Przedborski S. Time course and morphology of dopaminergic neuronal death caused by the neurotoxin 1-methyl-4- phenyl-1,2,3,6-tetrahydropyridine. Neurodegeneration 1995; 4(3): 257-69.

Fernandez-Alvarez A, Gomez-Sena L, Fabbiani MG, Budelli R, Abudara V. Endogenous presynaptic nitric oxide supports an anterograde signaling in the central nervous system. J Neurochem 2011; 118(4): 546-57.

Garthwaite J, Boulton CL. Nitric oxide signaling in the central nervous system. Annu Rev Physiol 1995; 57: 683-706.

Garthwaite J, Charles SL, Chess-Williams R. Endothelium-derived relaxing factor release on activation of NMDA receptors suggests role as intercellular messenger in the brain. Nature 1988; 336(6197): 385-8.

Garthwaite J, Garthwaite G, Palmer RM, Moncada S. NMDA receptor activation induces nitric oxide synthesis from arginine in rat brain slices. Eur J Pharmacol 1989; 172(4-5): 413-6.

Adamczyk A, Czapski GA, Kazmierczak A, Strosznajder JB. Effect of N-methyl-D- aspartate (NMDA) receptor antagonists on alpha-synuclein-evoked neuronal nitric oxide synthase activation in the rat brain. Pharmacol Rep 2009; 61(6): 1078-85.

East SJ, Garthwaite J. NMDA receptor activation in rat hippocampus induces cyclic GMP formation through the L-arginine- nitric oxide pathway. Neurosci Lett 1991; 123(1): 17-9.

Lei SZ, Pan ZH, Aggarwal SK, Chen HS, Hartman J, Sucher NJ, et al. Effect of nitric oxide production on the redox modulatory site of the NMDA receptor-channel complex. Neuron 1992; 8(6): 1087-99.

Manzoni O, Prezeau L, Marin P, Desagher S, Bockaert J, Fagni L. Nitric oxide-induced blockade of NMDA receptors. Neuron 1992; 8(4): 653-62.

Dawson VL, Dawson TM, London ED, Bredt DS, Snyder SH. Nitric oxide mediates glutamate neurotoxicity in primary cortical cultures. Proc Natl Acad Sci U S A 1991; 88(14): 6368-71.

Caldwell M, O'Neill M, Earley B, Leonard B. NG-Nitro-L-arginine protects against ischaemia-induced increases in nitric oxide and hippocampal neuro-degeneration in the gerbil. Eur J Pharmacol 1994; 260(2-3): 191-200.

Nowicki JP, Duval D, Poignet H, Scatton B. Nitric oxide mediates neuronal death after focal cerebral ischemia in the mouse. Eur J Pharmacol 1991; 204(3): 339-40.

Coleman JW. Nitric oxide in immunity and inflammation. Int Immunopharmacol 2001; 1(8): 1397-406.

Clancy RM, Amin AR, Abramson SB. The role of nitric oxide in inflammation and immunity. Arthritis Rheum 1998; 41(7):1141-51.

McCrabb GJ, Harding R. Role of nitric oxide in the regulation of cerebral blood flow in the ovine foetus. Clin Exp Pharmacol Physiol 1996; 23(10-11): 855-60.

Buchanan JE, Phillis JW. The role of nitric oxide in the regulation of cerebral blood flow. Brain Res 1993; 610(2): 248-55.

Yi J, Horky LL, Friedlich AL, Shi Y, Rogers JT, Huang X. L-arginine and Alzheimer's disease. Int J Clin Exp Pathol 2009; 2(3): 211-38.

Liu P, Fleete MS, Jing Y, Collie ND, Curtis MA, Waldvogel HJ, et al. Altered arginine metabolism in Alzheimer's disease brains. Neurobiol Aging 2014; 35(9): 1992-2003.

Virarkar M, Alappat L, Bradford PG, Awad AB. L-arginine and nitric oxide in CNS function and neurodegenerative diseases. Crit Rev Food Sci Nutr 2013; 53(11): 1157-67.

Ishida K, Shimizu H, Hida H, Urakawa S, Ida K, Nishino H. Argyrophilic dark neurons represent various states of neuronal damage in brain insults: some come to die and others survive. Neuroscience 2004; 125(3): 633-44.

Gallyas F, Kiglics V, Baracskay P, Juhasz G, Czurko A. The mode of death of epilepsy-induced "dark" neurons is neither necrosis nor apoptosis: an electron- microscopic study. Brain Res 2008; 1239: 207-15.

Kherani ZS, Auer RN. Pharmacologic analysis of the mechanism of dark neuron production in cerebral cortex. Acta Neuropathol 2008; 116(4): 447-52.

Ahmadpour SH, Haghir H. Diabetes mellitus type 1 induces dark neuron formation in the dentate gyrus: a study by Gallyas' method and transmission electron microscopy. Rom J Morphol Embryol 2011; 52(2): 575-9.

Cammermeyer J. I. An evaluation of the significance of the "dark" neuron. Ergeb Anat Entwicklungsgesch 1962; 36: 1-61.

Garman RH. The return of the dark neuron. A histological artifact complicating contemporary neurotoxicologic evaluation. Neurotoxicology 2006; 27(6): 1126.

Jortner BS. The return of the dark neuron. A histological artifact complicating contemporary neurotoxicologic evaluation. Neurotoxicology 2006; 27(4): 628-34.

Jafarian M, Rahimi S, Behnam F, Hosseini M, Haghir H, Sadeghzadeh B, et al. The effect of repetitive spreading depression on neuronal damage in juvenile rat brain.Neuroscience 2010; 169(1): 388-94.

Matheus FC, Aguiar AS, Castro AA, Villarinho JG, Ferreira J, Figueiredo CP, et al. Neuroprotective effects of agmatine in mice infused with a single intranasal administration of 1-methyl-4-phenyl- 1,2,3,6-tetrahydropyridine (MPTP). Behav Brain Res 2012; 235(2): 263-72.

Paxinos G, Watson C. The Rat Brain in Stereotaxic Coordinates: Hard Cover Edition. 6th ed. Waltham, MA: Academic Press; 2007.

Gundersen HJ, Bagger P, Bendtsen TF, Evans SM, Korbo L, Marcussen N, et al. The new stereological tools: disector, fractionator, nucleator and point sampled intercepts and their use in pathological research and diagnosis. APMIS 1988; 96(10): 857-81.

Salawu FK, Danburam A, Olokoba AB. Non-motor symptoms of Parkinson's disease: diagnosis and management. Niger J Med 2010; 19(2): 126-31.

Meissner W, Hill MP, Tison F, Gross CE, Bezard E. Neuroprotective strategies for Parkinson's disease: conceptual limits of animal models and clinical trials. Trends Pharmacol Sci 2004; 25(5): 249-53.

Blandini F, Greenamyre JT, Nappi G. The role of glutamate in the pathophysiology of Parkinson's disease. Funct Neurol 1996; 11(1): 3-15.

Rosa AO, Lin J, Calixto JB, Santos AR, Rodrigues AL. Involvement of NMDA receptors and L-arginine-nitric oxide pathway in the antidepressant-like effects of zinc in mice. Behav Brain Res 2003; 144(1- 2): 87-93.

Freitas AE, Moretti M, Budni J, Balen GO, Fernandes SC, Veronezi PO, et al. NMDA receptors and the L-arginine-nitric oxide- cyclic guanosine monophosphate pathway are implicated in the antidepressant-like action of the ethanolic extract from Tabebuia avellanedae in mice. J Med Food 2013; 16(11): 1030-8.

Ates-Alagoz Z, Adejare A. NMDA Receptor Antagonists for Treatment of Depression. Pharmaceuticals (Basel ) 2013; 6(4): 480-99.

Cherian L, Chacko G, Goodman C, Robertson CS. Neuroprotective effects of L- arginine administration after cortical impact injury in rats: dose response and time window. J Pharmacol Exp Ther 2003; 304(2): 617-23.

Lundblad C, Bentzer P. Effects of L- arginine on cerebral blood flow, microvascular permeability, number of perfused capillaries, and brain water content in the traumatized mouse brain. Microvasc Res 2007; 74(1): 1-8.

Garry PS, Ezra M, Rowland MJ, Westbrook J, Pattinson KT. The role of the nitric oxide pathway in brain injury and its treatment-- from bench to bedside. Exp Neurol 2015; 263: 235-43.

Cherian L, Chacko G, Goodman JC, Robertson CS. Cerebral hemodynamic effects of phenylephrine and L-arginine after cortical impact injury. Crit Care Med 1999; 27(11): 2512-7.

Jadeski LC, Lala PK. Nitric oxide synthase inhibition by N(G)-nitro-L-arginine methyl ester inhibits tumor-induced angiogenesis in mammary tumors. Am J Pathol 1999; 155(4): 1381-90.

Tripathi P, Misra MK. Therapeutic role of L-arginine on free radical scavenging system in ischemic heart diseases. Indian J Biochem Biophys 2009; 46(6): 498-502.

Dedkova EN, Blatter LA. Characteristics and function of cardiac mitochondrial nitric oxide synthase. J Physiol 2009; 587(Pt 4): 851-72.

Hazell AS, Itzhak Y, Liu H, Norenberg MD.1-Methyl-4-phenyl-1,2,3,6- tetrahydropyridine (MPTP) decreases glutamate uptake in cultured astrocytes. J Neurochem 1997; 68(5): 2216-9.

Lerman A, Burnett JC, Jr., Higano ST, McKinley LJ, Holmes DR, Jr. Long-term L- arginine supplementation improves small- vessel coronary endothelial function in humans. Circulation 1998; 97(21): 2123-8.

Kontos HA, Wei EP. Cerebral arteriolar dilations by KATP channel activators need L-lysine or L-arginine. Am J Physiol 1998; 274(3 Pt 2): H974-H981.

Iadecola C. Regulation of the cerebral microcirculation during neural activity: is nitric oxide the missing link? Trends Neurosci 1993; 16(6): 206-14.

Shin CY, Lee NI, Je HD, Kim JS, Sung JH, Kim DS, et al. Cardiovascular responses and nitric oxide production in cerebral ischemic rats. Arch Pharm Res 2002; 25(5): 697-703.

Condello S, Calabro E, Caccamo D, Curro M, Ferlazzo N, Satriano J, et al. Protective effects of agmatine in rotenone-induced damage of human SH-SY5Y neuroblastoma cells: fourier transform infrared spectroscopy analysis in a model of Parkinson's disease. Amino Acids 2012; 42(2-3): 775-81.

Martinez-Org, Fernandez-Frutos B, Gonzalez R,Fernandez-Lopez D,Uriguen L, Romero E, et al. Neuroprotective effect of L-arginine in a newborn rat model of acute severe asphyxia. Biol Neonate 2005; 88(4): 291-8.


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