open access

Vol 56, No 1 (2018)
Original paper
Submitted: 2017-08-01
Accepted: 2018-03-09
Published online: 2018-03-22
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Neuronal nitric oxide synthase phosphorylation induced by docosahexaenoic acid protects dopaminergic neurons in an experimental model of Parkinson’s disease

Hande Parlak1, Ayse Ozkan1, Sayra Dilmac2, Gamze Tanriover2, Ozlem Ozsoy1, Aysel Agar1
DOI: 10.5603/FHC.a2018.0005
·
Pubmed: 29577226
·
Folia Histochem Cytobiol 2018;56(1):27-37.
Affiliations
  1. Akdeniz University, Faculty of Medicine, Department of Physiology, Antalya, Turkey
  2. Akdeniz University, Faculty of Medicine, Department of Histology and Embryology, Antalya, Turkey, Antalya, Turkey

open access

Vol 56, No 1 (2018)
ORIGINAL PAPERS
Submitted: 2017-08-01
Accepted: 2018-03-09
Published online: 2018-03-22

Abstract

Introduction. Docosahexaenoic acid (DHA) has been shown to have beneficial effects on Parkinson’s disease
(PD). The aim of this study was to investigate if the DHA acts on neurons of substantia nigra (SN) by phosphorylation of neuronal nitric oxide synthase (nNOS) in an experimental mouse model of PD.
Material and methods. An experimental model of PD was created by intraperitoneal injections (4 × 20 mg/kg)
of the neurotoxin 1-methyl-phenyl-1, 2, 3, 6-tetrahydropyridine (MPTP). Three-month-old male C57BL/6 mice
were randomly divided into four groups as follows: control (C), DHA-treated (DHA), MPTP-injected (MPTP)
and DHA-treated and MPTP-injected (DHA + MPTP). DHA (36 mg/kg/day) was administered daily by gavage
for four weeks. Motor activity of the mice was evaluated with pole, locomotor activity and rotarod tests. Caspase-3
activity, nitrate/nitrite and 4-hydroxynonenal (4-HNE) levels were determined by spectrophotometric assays.
Immunohistochemistry was used to localize and assess the expressions of tyrosine hydroxylase (TH), nNOS and
phospho-nNOS (p-nNOS) in SN.
Results. An increased return and total down time in the MPTP group was observed in the pole test, while DHA
treatment decreased both parameters. The ambulatory activity, total distance and total locomotor activities were
decreased in the MPTP group, whereas they were increased by DHA treatment. MPTP-treated animals exhibited
shorter time on the rod test which was significantly increased by DHA treatment. DHA administration significantly
decreased 4-HNE and nitrate/nitrite levels of SN supernatants and protected the TH (+) dopaminergic
neurons of SN in the DHA + MPTP group compared to the MPTP group. DHA treatment significantly decreased
nNOS and increased p-nNOS immunoreactivities in the DHA + MPTP group compared to the MPTP group.
Conclusions. These results indicate that DHA treatment protects dopaminergic neurons in SN via increasing
nNOS serine 852 phosphorylation in the experimental mice model of PD.

Abstract

Introduction. Docosahexaenoic acid (DHA) has been shown to have beneficial effects on Parkinson’s disease
(PD). The aim of this study was to investigate if the DHA acts on neurons of substantia nigra (SN) by phosphorylation of neuronal nitric oxide synthase (nNOS) in an experimental mouse model of PD.
Material and methods. An experimental model of PD was created by intraperitoneal injections (4 × 20 mg/kg)
of the neurotoxin 1-methyl-phenyl-1, 2, 3, 6-tetrahydropyridine (MPTP). Three-month-old male C57BL/6 mice
were randomly divided into four groups as follows: control (C), DHA-treated (DHA), MPTP-injected (MPTP)
and DHA-treated and MPTP-injected (DHA + MPTP). DHA (36 mg/kg/day) was administered daily by gavage
for four weeks. Motor activity of the mice was evaluated with pole, locomotor activity and rotarod tests. Caspase-3
activity, nitrate/nitrite and 4-hydroxynonenal (4-HNE) levels were determined by spectrophotometric assays.
Immunohistochemistry was used to localize and assess the expressions of tyrosine hydroxylase (TH), nNOS and
phospho-nNOS (p-nNOS) in SN.
Results. An increased return and total down time in the MPTP group was observed in the pole test, while DHA
treatment decreased both parameters. The ambulatory activity, total distance and total locomotor activities were
decreased in the MPTP group, whereas they were increased by DHA treatment. MPTP-treated animals exhibited
shorter time on the rod test which was significantly increased by DHA treatment. DHA administration significantly
decreased 4-HNE and nitrate/nitrite levels of SN supernatants and protected the TH (+) dopaminergic
neurons of SN in the DHA + MPTP group compared to the MPTP group. DHA treatment significantly decreased
nNOS and increased p-nNOS immunoreactivities in the DHA + MPTP group compared to the MPTP group.
Conclusions. These results indicate that DHA treatment protects dopaminergic neurons in SN via increasing
nNOS serine 852 phosphorylation in the experimental mice model of PD.

Get Citation

Keywords

Parkinson’s disease; mice; protection; MPTP; DHA; nNOS phosphorylation; lipid peroxidation; caspase-3

About this article
Title

Neuronal nitric oxide synthase phosphorylation induced by docosahexaenoic acid protects dopaminergic neurons in an experimental model of Parkinson’s disease

Journal

Folia Histochemica et Cytobiologica

Issue

Vol 56, No 1 (2018)

Article type

Original paper

Pages

27-37

Published online

2018-03-22

DOI

10.5603/FHC.a2018.0005

Pubmed

29577226

Bibliographic record

Folia Histochem Cytobiol 2018;56(1):27-37.

Keywords

Parkinson’s disease
mice
protection
MPTP
DHA
nNOS phosphorylation
lipid peroxidation
caspase-3

Authors

Hande Parlak
Ayse Ozkan
Sayra Dilmac
Gamze Tanriover
Ozlem Ozsoy
Aysel Agar

References (62)
  1. Parkinson J. An essay on the shaking palsy. 1817. J Neuropsychiatry Clin Neurosci. 2002; 14(2): 223–36; discussion 222.
  2. Przedborski S. Pathogenesis of nigral cell death in Parkinson's disease. Parkinsonism Relat Disord. 2005; 11 Suppl 1: S3–S7.
  3. Chase A. Parkinson disease: Traumatic brain injury increases the risk of Parkinson disease. Nat Rev Neurol. 2015; 11(4): 184.
  4. Reeve A, Simcox E, Turnbull D. Ageing and Parkinson's disease: why is advancing age the biggest risk factor? Ageing Res Rev. 2014; 14: 19–30.
  5. Breydo L, Wu JW, Uversky VN. Α-synuclein misfolding and Parkinson's disease. Biochim Biophys Acta. 2012; 1822(2): 261–285.
  6. Verstraeten A, Theuns J, Van Broeckhoven C. Progress in unraveling the genetic etiology of Parkinson disease in a genomic era. Trends Genet. 2015; 31(3): 140–149.
  7. Schapira AHV. Mitochondria in the etiology of Parkinson's disease. Handb Clin Neurol. 2007; 83: 479–491.
  8. Dzamko N, Geczy CL, Halliday GM. Inflammation is genetically implicated in Parkinson's disease. Neuroscience. 2015; 302: 89–102.
  9. Su X, Maguire-Zeiss KA, Giuliano R, et al. Synuclein activates microglia in a model of Parkinson's disease. Neurobiol Aging. 2008; 29(11): 1690–1701.
  10. Ayala A, Muñoz MF, Argüelles S. Lipid peroxidation: production, metabolism, and signaling mechanisms of malondialdehyde and 4-hydroxy-2-nonenal. Oxid Med Cell Longev. 2014; 2014: 360438.
  11. Radi R. Peroxynitrite, a stealthy biological oxidant. J Biol Chem. 2013; 288(37): 26464–26472.
  12. El-Mlili N, Rodrigo R, Naghizadeh B, et al. Chronic hyperammonemia reduces the activity of neuronal nitric oxide synthase in cerebellum by altering its localization and increasing its phosphorylation by calcium-calmodulin kinase II. J Neurochem. 2008; 106(3): 1440–1449.
  13. Adak S, Santolini J, Tikunova S, et al. Neuronal nitric-oxide synthase mutant (Ser-1412 --> Asp) demonstrates surprising connections between heme reduction, NO complex formation, and catalysis. J Biol Chem. 2001; 276(2): 1244–1252.
  14. Rameau GA, Chiu LY, Ziff EB. Bidirectional regulation of neuronal nitric-oxide synthase phosphorylation at serine 847 by the N-methyl-D-aspartate receptor. J Biol Chem. 2004; 279(14): 14307–14314.
  15. Dyall SC, Michael-Titus AT. Neurological benefits of omega-3 fatty acids. Neuromolecular Med. 2008; 10(4): 219–235.
  16. Aras S, Tanriover G, Aslan M, et al. The role of nitric oxide on visual-evoked potentials in MPTP-induced Parkinsonism in mice. Neurochem Int. 2014; 72: 48–57.
  17. Jackson-Lewis V, Przedborski S. Protocol for the MPTP mouse model of Parkinson's disease. Nat Protoc. 2007; 2(1): 141–151.
  18. Przedborski S, Jackson-Lewis V, Yokoyama R, et al. Role of neuronal nitric oxide in 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-induced dopaminergic neurotoxicity. Proceedings of the National Academy of Sciences. 1996; 93(10): 4565–4571.
  19. Bousquet M, Saint-Pierre M, Julien C, et al. Beneficial effects of dietary omega-3 polyunsaturated fatty acid on toxin-induced neuronal degeneration in an animal model of Parkinson's disease. FASEB J. 2008; 22(4): 1213–1225.
  20. Surette ME. The science behind dietary omega-3 fatty acids. CMAJ. 2008; 178(2): 177–180.
  21. Luchtman DW, Meng Q, Wang X, et al. ω-3 fatty acid eicosapentaenoic acid attenuates MPP+-induced neurodegeneration in fully differentiated human SH-SY5Y and primary mesencephalic cells. J Neurochem. 2013; 124(6): 855–868.
  22. Ozsoy O, Tanriover G, Derin N, et al. The effect of docosahexaenoic Acid on visual evoked potentials in a mouse model of Parkinson's disease: the role of cyclooxygenase-2 and nuclear factor kappa-B. Neurotox Res. 2011; 20(3): 250–262.
  23. Kobayashi T, Araki T, Itoyama Y, et al. Effects of L-dopa and bromocriptine on haloperidol-induced motor deficits in mice. Life Sci. 1997; 61(26): 2529–2538.
  24. Ogawa N, Mizukawa K, Hirose Y, et al. MPTP-induced parkinsonian model in mice: biochemistry, pharmacology and behavior. Eur Neurol. 1987; 26 Suppl 1: 16–23.
  25. Kayir H, Uzbay IT. Evidence for the role of nitric oxide in caffeine-induced locomotor activity in mice. Psychopharmacology (Berl). 2004; 172(1): 11–15.
  26. Rozas G, Guerra MJ, Labandeira-Garcı́a JL. An automated rotarod method for quantitative drug-free evaluation of overall motor deficits in rat models of parkinsonism. Brain Research Protocols. 1997; 2(1): 75–84.
  27. Rozas G, López-Martín E, Guerra MJ, et al. The overall rod performance test in the MPTP-treated-mouse model of Parkinsonism. J Neurosci Methods. 1998; 83(2): 165–175.
  28. Bradford MM. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976; 72: 248–254.
  29. Schober A. Classic toxin-induced animal models of Parkinson's disease: 6-OHDA and MPTP. Cell Tissue Res. 2004; 318(1): 215–224.
  30. Little SJ, Lynch MA, Manku M, et al. Docosahexaenoic acid-induced changes in phospholipids in cortex of young and aged rats: a lipidomic analysis. Prostaglandins Leukot Essent Fatty Acids. 2007; 77(3-4): 155–162.
  31. Samadi P, Grégoire L, Rouillard C, et al. Docosahexaenoic acid reduces levodopa-induced dyskinesias in 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine monkeys. Ann Neurol. 2006; 59(2): 282–288.
  32. Jiang Lh, Shi Y, Wang Ls, et al. The influence of orally administered docosahexaenoic acid on cognitive ability in aged mice. J Nutr Biochem. 2009; 20(9): 735–741.
  33. Hacioglu G, Agar A, Yargicoglu P. The role of docosahexaenoic acid on visual evoked potentials in one kidney-one clip hypertension. Acta Ophthalmol Scand. 2006; 84(4): 488–494.
  34. Tanriover G, Seval-Celik Y, Ozsoy O, et al. The effects of docosahexaenoic acid on glial derived neurotrophic factor and neurturin in bilateral rat model of Parkinson's disease. Folia Histochem Cytobiol. 2010; 48(3): 434–441.
  35. Luchtman DW, Meng Q, Song C. Ethyl-eicosapentaenoate (E-EPA) attenuates motor impairments and inflammation in the MPTP-probenecid mouse model of Parkinson's disease. Behav Brain Res. 2012; 226(2): 386–396.
  36. Tekin I, Roskoski R, Carkaci-Salli N, et al. Complex molecular regulation of tyrosine hydroxylase. J Neural Transm (Vienna). 2014; 121(12): 1451–1481.
  37. Pickrell AM, Pinto M, Hida A, et al. Striatal Dysfunctions Associated with Mitochondrial DNA Damage in Dopaminergic Neurons in a Mouse Model of Parkinson's Disease. Journal of Neuroscience. 2011; 31(48): 17649–17658.
  38. Zhu Y, Zhang J, Zeng Y. Overview of tyrosine hydroxylase in Parkinson's disease. CNS Neurol Disord Drug Targets. 2012; 11(4): 350–358.
  39. Xiong ZK, Lang J, Xu G, et al. Excessive levels of nitric oxide in rat model of Parkinson's disease induced by rotenone. Exp Ther Med. 2015; 9(2): 553–558.
  40. Steinert JR, Chernova T, Forsythe ID. Nitric oxide signaling in brain function, dysfunction, and dementia. Neuroscientist. 2010; 16(4): 435–452.
  41. Guo S, Yan J, Yang T, et al. Protective effects of green tea polyphenols in the 6-OHDA rat model of Parkinson's disease through inhibition of ROS-NO pathway. Biol Psychiatry. 2007; 62(12): 1353–1362.
  42. Kaur B, Prakash A. Ceftriaxone attenuates glutamate-mediated neuro-inflammation and restores BDNF in MPTP model of Parkinson's disease in rats. Pathophysiology. 2017; 24(2): 71–79.
  43. Hayashi Y, Nishio M, Naito Y, et al. Regulation of neuronal nitric-oxide synthase by calmodulin kinases. J Biol Chem. 1999; 274(29): 20597–20602.
  44. Komeima K, Hayashi Y, Naito Y, et al. Inhibition of neuronal nitric-oxide synthase by calcium/ calmodulin-dependent protein kinase IIalpha through Ser847 phosphorylation in NG108-15 neuronal cells. J Biol Chem. 2000; 275(36): 28139–28143.
  45. Sofic, E., A. Sapcanin, I. Tahirovic, , Antioxidant capacity in postmortem brain tissues of Parkinson's and Alzheimer's diseases. J Neural Transm Suppl, 2006(71): p. : 39–43.
  46. Chen X, Guo C, Kong J. Oxidative stress in neurodegenerative diseases. Neural Regen Res. 2012; 7(5): 376–385.
  47. Poljsak B. J.P., Methodology for oxidative state detection in biological systems, in Handbook of Free Radicals: Formation, Types and Effects, S.V. Kozyrev D, Editor. 2010, Nova Science: New York, NY. : USA.
  48. Birben E, Sahiner UM, Sackesen C, et al. Oxidative stress and antioxidant defense. World Allergy Organ J. 2012; 5(1): 9–19.
  49. Reed TT. Lipid peroxidation and neurodegenerative disease. Free Radic Biol Med. 2011; 51(7): 1302–1319.
  50. Liu, W., M. Kato, A.A. Akhand, , 4-hydroxynonenal induces a cellular redox status-related activation of the caspase cascade for apoptotic cell death. J Cell Sci, 2000. 113 ( Pt 4): p. : 635–41.
  51. Qin Z, Hu D, Han S, et al. Effect of 4-hydroxy-2-nonenal modification on alpha-synuclein aggregation. J Biol Chem. 2007; 282(8): 5862–5870.
  52. Lv C, Hong T, Yang Z, et al. Effect of Quercetin in the 1-Methyl-4-phenyl-1, 2, 3, 6-tetrahydropyridine-Induced Mouse Model of Parkinson's Disease. Evid Based Complement Alternat Med. 2012; 2012: 928643.
  53. Ghosh A, Kanthasamy A, Joseph J, et al. Anti-inflammatory and neuroprotective effects of an orally active apocynin derivative in pre-clinical models of Parkinson's disease. J Neuroinflammation. 2012; 9: 241.
  54. Takahashi M, Tsuboyama-Kasaoka N, Nakatani T, et al. Fish oil feeding alters liver gene expressions to defend against PPARalpha activation and ROS production. Am J Physiol Gastrointest Liver Physiol. 2002; 282(2): G338–G348.
  55. Dehmer T, Lindenau J, Haid S, et al. Deficiency of inducible nitric oxide synthase protects against MPTP toxicity in vivo. J Neurochem. 2000; 74(5): 2213–2216.
  56. Moon HE, Paek SHa. Mitochondrial Dysfunction in Parkinson's Disease. Exp Neurobiol. 2015; 24(2): 103–116.
  57. Chai J, Shi Y. Apoptosome and inflammasome: conserved machineries for caspase activation. National Science Review. 2014; 1(1): 101–118.
  58. Gaschler MM, Stockwell BR. Lipid peroxidation in cell death. Biochem Biophys Res Commun. 2017; 482(3): 419–425.
  59. Stillwell W, Wassall S. Docosahexaenoic acid: membrane properties of a unique fatty acid. Chemistry and Physics of Lipids. 2003; 126(1): 1–27.
  60. German OL, Monaco S, Agnolazza DL, et al. Retinoid X receptor activation is essential for docosahexaenoic acid protection of retina photoreceptors. J Lipid Res. 2013; 54(8): 2236–2246.
  61. Bousquet M, Gue K, Emond V, et al. Transgenic conversion of omega-6 into omega-3 fatty acids in a mouse model of Parkinson's disease. J Lipid Res. 2011; 52(2): 263–271.
  62. Chalon S, Delion-Vancassel S, Belzung C, et al. Dietary Fish Oil Affects Monoaminergic Neurotransmission and Behavior in Rats. The Journal of Nutrition. 1998; 128(12): 2512–2519.

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