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. 2013 Jul 19;15(3):115–122. doi: 10.1179/174329210X12650506623447

Involvement of the mitochondrial apoptotic pathway and nitric oxide synthase in dopaminergic neuronal death induced by 6-hydroxydopamine and lipopolysaccharide

Sarika Singh 1, Sachin Kumar 2, Madhu Dikshit 3
PMCID: PMC7067325  PMID: 20594414

Abstract

The primary pathology in Parkinson's disease patients is significant loss of dopaminergic neurons in the substantia nigra through multiple mechanisms. We previously have demonstrated the involvement of nitric oxide (NO) in the dopaminergic neurodegeneration induced by 6-hydroxydopamine (6-OHDA) and lipopolysaccharide (LPS) in rats. The present study was undertaken to investigate further the role of NO in the mitochondria-mediated apoptosis of dopaminergic neurons during the early time period after administration of 6-OHDA and LPS. Measurement of dopamine and its metabolites, TH immunolabeling, cytochrome-c release, mitochondrial complex-I and caspase-3 activity assessment was performed in both the 6-OHDA- and LPS-induced experimental models of Parkinson's disease. Significant decreases in dopamine, 3,4-dihydroxyphenylacetic acid (DOPAC), homovanillic acid (HVA), tyrosine hydroxylase (TH) immunolabeling and mitochondrial complex-I activity were observed, with increase in cytochrome-c release and caspase-3 activation. Dopmaine and its metabolite levels, mitochondrial complex-I activity and caspase-3 activity were significantly reversed with treatment of the NOS inhibitor, L-NAME. The reduction in the extent of cytochrome-c release responded variably to NOS inhibition in both the models. The results obtained suggest that NO contributes to mitochondria-mediated neuronal apoptosis in the dopaminergic neurodegeneration induced by 6-OHDA and LPS in rats.

Keywords: 6-HYDROXYDOPAMINE, LIPOPOLYSACCHARIDE, NITRIC OXIDE, APOPTOSIS, MITOCHONDRIA, CASPASE

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References

  • 1.Schapira AH, Cooper JM, Dexter D, Clark JB, Jenner P, Marsden CD. Mitochondrial complex I deficiency in Parkinson's disease. J Neurochem 1990; 54: 823–827. [DOI] [PubMed] [Google Scholar]
  • 2.de la Monte SM, Sohn YK, Ganju N, Wands JR. P53- and CD95-associated apoptosis in neurodegenerative diseases. Lab Invest 1998; 78: 401–411. [PubMed] [Google Scholar]
  • 3.Hartmann A, Hunot S, Michel PP et al. Caspase-3: a vulnerability factor and final effector in apoptotic death of dopaminergic neurons in Parkinson's disease. Proc Natl Acad Sci USA 2000; 97: 2875–2880. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 4.Mogi M, Togari A, Kondo T et al. Caspase activities and tumor necrosis factor receptor R1 (p55) level are elevated in the substantia nigra from parkinsonian brain. J Neural Transm 2000; 107: 335–341. [DOI] [PubMed] [Google Scholar]
  • 5.Hartmann A, Troadec JD, Hunot S et al. Caspase-8 is an effector in apoptotic death of dopaminergic neurons in Parkinson's disease, but pathway inhibition results in neuronal necrosis. J Neurosci 2001; 21: 2247–2255. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 6.Lee DH, Han YS, Han ES, Bang H, Lee CS. Differential involvement of intracellular Ca2* in 1-methyl-4-phenylpyridinium- or 6-hydroxy-dopamine-induced cell viability loss in PC12 cells. Neurochem Res 2006; 31: 851–860. [DOI] [PubMed] [Google Scholar]
  • 7.Singh S, Dikshit M. Apoptotic neuronal death in Parkinson's disease: involvement of nitric oxide. Brain Res Rev 2007; 54: 233–250. [DOI] [PubMed] [Google Scholar]
  • 8.Takai N, Nakanishi H, Tanabe K et al. Involvement of caspase-like proteinases in apoptosis of neuronal PC12 cells and primary cultured microglia induced by 6-hydroxydopamine. J Neurosci Res 1998; 54: 214–222. [DOI] [PubMed] [Google Scholar]
  • 9.Fall CP, Bennett JP Jr. Characterization and time course of MPP*-induced apoptosis in human SH-SY5Y neuroblastoma cells. J Neurosci Res 1999; 55: 620–628. [DOI] [PubMed] [Google Scholar]
  • 10.Blum D, Torch S, Lambeng N et al. Molecular pathways involved in the neurotoxicity of 6-0HDA, dopamine and MPTP: contribution to the apoptotic theory in Parkinson's disease. Prog Neurobiol 2001; 65: 135–172. [DOI] [PubMed] [Google Scholar]
  • 11.Von Coelln R, Kugler S, Bahr M, Weller M, Dichgans J, Schulz JB. Rescue from death but not from functional impairment: caspase inhibition protects dopaminergic cells against 6-hydroxydopamine-induced apoptosis but not against the loss of their terminals. J Neurochem 2001; 77: 263–273. [DOI] [PubMed] [Google Scholar]
  • 12.Gomez C, Reiriz J, Pique M, Gil J, Ferrer I, Ambrosio S. Low concentrations of 1-methyl-4-phenylpyridinium ion induce caspase-mediated apoptosis in human SH-SY5Y neuroblastoma cells. J Neurosci Res 2001; 63: 421–428. [DOI] [PubMed] [Google Scholar]
  • 13.Han BS, Hong HS, Choi WS, Markelonis GJ, Oh TH, Oh YJ. Caspase-dependent and independent cell death pathways in primary cultures of mesencephalic dopaminergic neurons after neurotoxin treatment. J Neurosci 2003; 23: 5069–5078. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 14.Noelker C, Bacher M, Gocke P et al. The flavanoide caffeic acid phenethyl ester blocks 6-hydroxydopamine-induced neurotoxicity. Neurosci Lett 2005; 383: 39–43. [DOI] [PubMed] [Google Scholar]
  • 15.Blum D, Wu Y, Nissou MF, Arnaud S, Alim Louis B, Verna JM. p53 and Bax activation in 6-hydroxydopamine-induced apoptosis in PC12 cells. Brain Res 1997; 751: 139–142. [DOI] [PubMed] [Google Scholar]
  • 16.Jordan J, Galindo MF, Tomero D, Gonzalez-Garcia C, Cena V. Bc1-xl blocks mitochondrial multiple conductance channel activation and inhibits 6-0HDA-induced death in SH-SY5Y cells. J Neurochem 2004; 89: 124–133. [DOI] [PubMed] [Google Scholar]
  • 17.Cutillas B, Espejo M, Gil J, Ferrer I, Ambrosio S. Caspase inhibition protects nigral neurons against 6-0HDA-induced retrograde degeneration. Neuroreport 1999; 10: 2605–2608. [DOI] [PubMed] [Google Scholar]
  • 18.Raghavan SA, Dikshit M. Vascular regulation by the L-arginine metabolites, nitric oxide and agmatine. Pharmacol Res 2004; 49: 397–414. [DOI] [PubMed] [Google Scholar]
  • 19.Kavya R, Saluja R, Singh S, Dikshit M. Nitric oxide synthase regulation and diversity: implications in Parkinson's disease. Nitric Oxide 2006; 15: 280–294. [DOI] [PubMed] [Google Scholar]
  • 20.Duncan AJ, Heales SJ. Nitric oxide and neurological disorders. Mol Aspects Med 2005; 26: 67–96. [DOI] [PubMed] [Google Scholar]
  • 21.Zhang L, Dawson VL, Dawson TM. Role of nitric oxide in Parkinson's disease. Pharmacol Ther 2006; 109: 33–41. [DOI] [PubMed] [Google Scholar]
  • 22.Heales SJ, Bolanos JP, Stewart VC, Brookes PS, Land JM, Clark JB. Nitric oxide, mitochondria and neurological disease. Biochim Biophys Acta 1999; 1410: 215–228. [DOI] [PubMed] [Google Scholar]
  • 23.Barthwal MK, Srivastava N, Dikshit M. Role of nitric oxide in a progressive neurodegeneration model of Parkinson's disease in the rat. Redox Report 2001; 6: 297–302. [DOI] [PubMed] [Google Scholar]
  • 24.Singh S, Das T, Ravindran A et al. Involvement of nitric oxide in neurodegeneration: a study on the experimental models of Parkinson's disease Redox Report 2005; 10: 103–109. [DOI] [PubMed] [Google Scholar]
  • 25.Paxinos G, Watson C. The Rat Brain in Stereotaxic Coordinates, 2nd edn. New York: Academic Press, 1982; 83-97. [Google Scholar]
  • 26.Yokoyama C, Okamura H. Self-injurious behavior and dopaminergic neuron system in neonatal 6-hydroxydopamine-lesioned rat: 1. Dopaminergic neurons and receptors. J Pharmacol Exp Ther 1997; 280: 1016–1030. [PubMed] [Google Scholar]
  • 27.Rajapakse N, Shimizu K, Payne M, Busija D. Isolation and characterization of intact mitochondria from neonatal rat brain. Brain Res Protocol 2001; 8: 176–183. [DOI] [PubMed] [Google Scholar]
  • 28.Fujimura M, Morita-Fujimura Y, Murakami K, Kawase M, Chan PH. Cytosolic redistribution of cytochrome c after transient focal cerebral ischemia in rats. J Cerebr Blood Flow Metab 1998; 18: 1239–1247. [DOI] [PubMed] [Google Scholar]
  • 29.Heneka MT, Sastre M, Dumitrescu-Ozimek L et al. Focal glial activation coincides with increased BACE1 activation and precedes amyloid plaque deposition in APP[V7171] transgenic mice. J Neuroinflamm 2005; 2: 22. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 30.Cid C, Alvarez-Cermeno JC, Regidor I, Plaza J, Salinas M, Alcazar A. Caspase inhibitors protect against neuronal apoptosis induced by cerebrospinal fluid from multiple sclerosis patients. J Neuroimmunol 2003; 136: 119–124. [DOI] [PubMed] [Google Scholar]
  • 31.Castano A, Herrera AJ, Cano J, Machado A. Lipopolysaccharide intranigral injection induces inflammatory reaction and damage in nigrostriatal dopaminergic system. J Neurochem 1998; 70: 1584–1592. [DOI] [PubMed] [Google Scholar]
  • 32.Sanchez-Pemaute R, Ferree A, Cooper O, Yu M, Brownell AL, Isacson O. Selective COX-2 inhibition prevents progressive dopamine neuron degeneration in a rat model of Parkinson's disease. J Neuroinflamm 2004; 1: 6. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 33.Chung HT, Pae HO, Choi BM, Billiar TR, Kim YM. Nitric oxide as a bioregulator of apoptosis. Biochem Biophys Res Commun 2001; 282: 1075–1079. [DOI] [PubMed] [Google Scholar]
  • 34.Kim PK, Zamora R, Petrosko P, Billiar TR. The regulatory role of nitric oxide in apoptosis. Int Immunopharmacol 2001; 1: 1421–1441. [DOI] [PubMed] [Google Scholar]
  • 35.Paucard A, Palmier B, Croci N, Taillieu F, Plotkine M, Margaill I. Biphasic modulation by nitric oxide of caspase activation due to malonate injection in rat striatum. Eur J Pharmacol 2004; 483: 259–265. [DOI] [PubMed] [Google Scholar]
  • 36.Kitamura Y, Inden M, Miyamura A, Kakimura J, Taniguchi T, Shimohama S. Possible involvement of both mitochondria- and endoplasmic reticulum-dependent caspase pathways in rotenone-induced apoptosis in human neuroblastoma SH-SY5Y cells. Neurosci Lett 2002; 333: 25–28. [DOI] [PubMed] [Google Scholar]
  • 37.Hanrott K, Gudmunsen L, O'Neill MJ, Wonnacott S. 6-Hydroxydopamine-induced apoptosis is mediated via extracellular auto-oxidation and caspase 3-dependent activation of protein kinase Cdelta. J Biol Chem 2006; 281: 5373–5382. [DOI] [PubMed] [Google Scholar]
  • 38.Navarro A, Boveris A, Bandez MJ et al. Human brain cortex: mitochondrial oxidative damage and adaptive response in Parkinson disease and in dementia with Lewy bodies. Free Radic Biol Med 2009; 46: 1574–1580. [DOI] [PubMed] [Google Scholar]
  • 39.Brown GC, Borutaite V. Inhibition of mitochondrial respiratory complex I by nitric oxide, peroxynitrite and S-nitrosothiols. Biochim Biophys Acta 2004; 1658: 11 49. [DOI] [PubMed] [Google Scholar]
  • 40.Ahsen Von 0, Waterhouse NJ, Kuwana T, Newmeyer DD, Green DR. The ‘harmless’ release of cytochrome c. Cell Death Diff 2000; 7: 1192–1199. [DOI] [PubMed] [Google Scholar]
  • 41.Donovan M, Cotter TG. Control of mitochondrial integrity by Bc1-2 family members and caspase-independent cell death. Biochim Biophys Acta 2004; 1644: 133–147. [DOI] [PubMed] [Google Scholar]
  • 42.Munno I, Pellegrino NM, Marcuccio C, Conrotto L, Jirillo E, Covelli V. Neurological damage mediated by cytokines. Acta Neurol (Napoli) 1992; 14: 81–89. [PubMed] [Google Scholar]
  • 43.Ashe PC, Berry MD. Apoptotic signaling cascades. Prog Neuropsychopharmacol Biol Psychiatry 2003; 27: 199–214. [DOI] [PubMed] [Google Scholar]

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