The Cerebellum

, Volume 8, Issue 3, pp 366–372

Increase in Cerebellar Neurotrophin-3 and Oxidative Stress Markers in Autism

  • Elizabeth M. Sajdel-Sulkowska
  • Ming Xu
  • Noriyuki Koibuchi
Article

Abstract

Autism is a neurodevelopmental disorder characterized by social and language deficits, ritualistic–repetitive behaviors and disturbance in motor functions. Data of imaging, head circumference studies, and Purkinje cell analysis suggest impaired brain growth and development. Both genetic predisposition and environmental triggers have been implicated in the etiology of autism, but the underlying cause remains unknown. Recently, we have reported an increase in 3-nitrotyrosine (3-NT), a marker of oxidative stress damage to proteins in autistic cerebella. In the present study, we further explored oxidative damage in the autistic cerebellum by measuring 8-hydroxydeoxyguanosine (8-OH-dG), a marker of DNA modification, in a subset of cases analyzed for 3-NT. We also explored the hypothesis that oxidative damage in autism is associated with altered expression of brain neurotrophins critical for normal brain growth and differentiation. The content of 8-OH-dG in cerebellar DNA isolated by the proteinase K method was measured using an enzyme-linked immunosorbent assay (ELISA); neurotrophin-3 (NT-3) levels in cerebellar homogenates were measured using NT-3 ELISA. Cerebellar 8-OH-dG showed trend towards higher levels with the increase of 63.4% observed in autism. Analysis of cerebellar NT-3 showed a significant (p = 0.034) increase (40.3%) in autism. Furthermore, there was a significant positive correlation between cerebellar NT-3 and 3-NT (r = 0.83; p = 0.0408). These data provide the first quantitative measure of brain NT-3 and show its increase in the autistic brain. Altered levels of brain NT-3 are likely to contribute to autistic pathology not only by affecting brain axonal targeting and synapse formation but also by further exacerbating oxidative stress and possibly contributing to Purkinje cell abnormalities.

Keywords

Oxidative stress 8-OH-dG 3-nitrotyrosine (3-NT) Neurotrophin-3 (NT-3) Autism Cerebellum 

References

  1. 1.
    Bokara KK, Brown E, McCormick R, Yallapragada PR, Rajanna S, Bettaiya R (2008) Lead-induced increase in antioxidant enzymes and lipid peroxidation products in developing rat brain. Biometals 21:9–16PubMedCrossRefGoogle Scholar
  2. 2.
    Winham GC, Zhang L, Gunier R, Croen LA, Grether JK (2006) Autism spectrum disorders in relation to distribution of hazardous air pollutants in the San Francisco bay area. Environ Health Perspect 114:1438–1444CrossRefGoogle Scholar
  3. 3.
    Palmer RF, Blanchard S, Wood R (2008) Proximity to point sources of environmental mercury release as a predictor of autism prevalence. Health Place 15(1):18–24PubMedCrossRefGoogle Scholar
  4. 4.
    D’Amelio M, Ricci I, Sacco R, Liu X, D’Agruma L, Muscarella LA et al (2005) Paraoxonase gene variants are associated with autism in North America, but not in Italy: possible regional specificity in gene–environment interactions. Mol Psychiatry 10:1006–1016PubMedCrossRefGoogle Scholar
  5. 5.
    Kimura-Kuroda J, Nagata I, Kuroda Y (2007) Disrupting effect of hydroxyl-polychlorinated biphenyl (PCB) congeners on neuronal development of cerebellar Purkinje cells: a possible causal factor for the developmental brain disorders. Chemosphere 67:S412–S420PubMedCrossRefGoogle Scholar
  6. 6.
    Brown GE, Jones SD, MacKewn AS, Plank EJ (2008) An exploration of possible pre- and postnatal correlates of autism: a pilot survey. Psychol Rep 102:273–282PubMedCrossRefGoogle Scholar
  7. 7.
    Sultana R, Perluigi M, Butterfield DA (2006) Protein oxidation, lipid peroxidation in brain of subjects with Alzheimer’s disease: insights into mechanism of neurodegeneration from redox proteomics. Antioxid Redox Signal 8:2021–2037PubMedCrossRefGoogle Scholar
  8. 8.
    Neumann H, Hazen L, Weinstein J, Mehl RA, Chin JW (2008) Genetically encoding protein oxidative damage. J Am Chem Soc 130:4028–4033PubMedCrossRefGoogle Scholar
  9. 9.
    Sajdel-Sulkowska EM, Lipinski B, Windom H, Audhya T, McGinnis W (2008) Oxidative stress in autism: cerebellar 3 nitrotyrosine levels. Am J Biochem Biotechnol 4:73–84CrossRefGoogle Scholar
  10. 10.
    Lovell MA, Gabbita SP, Markesbery WR (1999) Increased DNA oxidation and decreased levels of repair products in Alzheimer’s disease ventricular CSF. J Neurochem 72:771–776PubMedCrossRefGoogle Scholar
  11. 11.
    Lee SH, Kim I, Chung BC (2007) Increased urinary level of oxidized nucleosides in patients with mild-to-moderate Alzheimer’s disease. Clin Biochem 40:936–938PubMedCrossRefGoogle Scholar
  12. 12.
    Sato S, Mizuno Y, Hattori N (2005) Urinary 8-hydroxydeoxyguanosine levels as a biomarker for progression of Parkinson disease. Neurology 64:1081–1083PubMedGoogle Scholar
  13. 13.
    Fukuda M, Yamaguchi H, Yamamoto H, Aminaka M, Murakami H, Kamiyama N et al (2008) The evaluation of oxidative damage in children with brain damage using 8-hydroxydeoxyguanosine levels. Brain Dev 30:131–136PubMedCrossRefGoogle Scholar
  14. 14.
    Nishioka N, Arnold SE (2004) Evidence for oxidative DNA damage in the hippocampus of elderly patients with chronic schizophrenia. Am J Geriatr Psychiatry 12:167–175PubMedGoogle Scholar
  15. 15.
    Ming X, Stein TP, Brimacombe M, Johnson WG, Lambert GH, Wagner GC (2005) Increased excretion of lipid peroxidation biomarker in autism. Prostaglandins Leukot Essent Fat Acids 73:379–384CrossRefGoogle Scholar
  16. 16.
    Courchesne E, Karns CM, Davis HR, Ziccardi R, Carper RA, Tigue ZD (2001) Unusual brain growth patterns in early life in patients with autistic disorder: an MRI study. Neurology 57:245–254PubMedGoogle Scholar
  17. 17.
    Whitney ER, Kemper TL, Bauman ML, Rosene DL, Blatt GJ (2008) Cerebellar Purkinje cells are reduced in a subpopulation of autistic brains: a stereological experiment using calbindin-D28k. Cerebellum 7(3):406–416PubMedCrossRefGoogle Scholar
  18. 18.
    Chao SL, Moss JM, Harry GJ (2007) Lead-induced alterations of apoptosis and neurotrophic factor mRNA in the developing rat cortex, hippocampus, and cerebellum. J Biochem Mol Toxicol 21:265–272PubMedCrossRefGoogle Scholar
  19. 19.
    Marx CE, Vance BJ, Jarskog LF, Chescheir NC, Gilmore JH (1999) Nerve growth factor, brain derived neurotrophic factor and neurotrophin-3 levels in human amniotic fluid. Am J Obstet Gynecol 181:1225–1230PubMedCrossRefGoogle Scholar
  20. 20.
    Courchesne E, Carper R, Akshoomoff N (2003) Evidence of brain overgrowth in the first year life in autism. JAMA 290:393–394CrossRefGoogle Scholar
  21. 21.
    Rivto ER, Freeman BJ, Scheibel AB, Duong T, Robinson H, Guthrie D, Ritvo A (1986) Lower Purkinje cell counts in the cerebella of four autistic subjects: initial findings of the UCLA-NSac Autopsy Research Report. Am. J. Psychiatry 143:862–866Google Scholar
  22. 22.
    Courchesne E (1991) Neuroanatomic imaging in autism. Pediatrics 87:781–790PubMedGoogle Scholar
  23. 23.
    Kemper TL, Bauman ML (1993) The contribution of neuropathologic studies to the understanding of autism. Neurol Clin 11:175–187PubMedGoogle Scholar
  24. 24.
    Das KP, Chao SL, White LD, Haines WT, Harry GJ, Tilson HA, Barone S Jr (2001) Differential patterns of nerve growth factor, brain-derived neurotrophic factor and neurotrophin-3 mRNA and protein levels in developing regions of rat brain. Neuroscience 103:739–761PubMedCrossRefGoogle Scholar
  25. 25.
    Li S, Qiu F, Xu A, Price SM, Xiang M (2004) Barhl1 regulates migration and survival of cerebellar granule cells by controlling expression of neurotrophin-3 gene. J Neuroscience 24:3104–3114CrossRefGoogle Scholar
  26. 26.
    Kawakami H, Nitta A, Matsuyama Y, Kamiya M, Satake K, Sato K et al (2000) Increase in neurotrophin-3 expression followed by Purkinje cell degeneration in the adult rat cerebellum after spinal cord transaction. J Neurosci Res 62:668–674PubMedCrossRefGoogle Scholar
  27. 27.
    Helbock HJ, Beckma KB, Ames BN (1999) 8-Hydroxydeoxyguanosine and 8 hydroxyguanine as biomarkers of oxidative stress. Method Enzymol 300:156–166CrossRefGoogle Scholar
  28. 28.
    Bershtein LM, Poroshina LVV, TE TEV (2005) Content of 8-hydroxy-2-deoxyguanosine in steroid receptor positive and receptor-negative breast cancer cells. Bull Exp Biol Med 140:88–91PubMedCrossRefGoogle Scholar
  29. 29.
    Miller MW, Mooney SM (2004) Chronic exposure to ethanol alters neurotrophin content in the basal forebrain–cortex system in the mature rat: effects on autocrine–paracrine mechanisms. J Neurobiol 60:490–498PubMedCrossRefGoogle Scholar
  30. 30.
    Abdollahi M, Ranjbar A, Shadnia S, Nikfar S, Rezaiee A (2004) Pesticides and oxidative stress: a review. Med Sci Monit 10:141–147Google Scholar
  31. 31.
    Mutter J, Naumann J, Schneider R, Wlach H, Haley B (2005) Mercury and autism: accelerating evidence? Neuro Endocrinol Lett 26:439–446PubMedGoogle Scholar
  32. 32.
    Mutter J, Naumann J, Walach H, Daschner F (2005) Amalgam risk assessment with coverage of references up to 2005. Gesundheitswesen 67:204–216PubMedCrossRefGoogle Scholar
  33. 33.
    Bradstreet J, Geier DA, Kartzinel JJ, Adams JB, Geier MR (2003) A case–control study of mercury burden in children with autistic disorders. J Am Phys Surg 8:76–80Google Scholar
  34. 34.
    Serajee FJ, Nabi R, Zhong H, Huq M (2004) Polymorhism in xenobiotic metabolism genes and autism. J Child Neurol 19:413–417PubMedGoogle Scholar
  35. 35.
    Zoroglu SS, Armutcu F, Ozen S, Gurel A, Sivasli E, Yetkin O et al (2004) Increased oxidative stress and altered activities of erythrocyte free radical scavenging enzymes in autism. Eur Arch Psychiatr Clin Neurosci 254:143–147Google Scholar
  36. 36.
    Yorbik O, Sayal A, Akay C, Akbiyik DI, Sohmen T (2002) Investigation of antioxidant enzymes in children with autistic disorder. Prostaglandins Leukot Essent Fat Acids 67:341–343CrossRefGoogle Scholar
  37. 37.
    James SJ, Cutler P, Melnyk S, Jernigan S, Janak L, Gaylor DW et al (2004) Metabolic biomarkers of increased oxidative stress and impaired methylation capacity in children with autism. Am J Clin Nutr 80:1611–1617PubMedGoogle Scholar
  38. 38.
    James SJ, Melny S, Jernigan S, Cleves MA, Halsted CH, Wong DH et al (2006) Metabolic endotype and related genotypes are associated with oxidative stress in children with autism. Am J Med Genet B Neuropsychiatry Genet 141:947–956CrossRefGoogle Scholar
  39. 39.
    Yang IA, Fong KM, Zimmerman PV, Holgate ST, Holloway JW (2008) Genetic susceptibility to the respiratory effects of air pollution. Thorax 63:555–563PubMedGoogle Scholar
  40. 40.
    Buyske S, Williams TA, Mars AE, Stenroos ES, Ming SX, Wang R et al (2006) Analysis of case-parent trios at a locus with a deletion allele: association of GSTM1 with autism. BMC 7:8Google Scholar
  41. 41.
    Nelson PG, Kuddo T, Song EY, Dambrosia JM, Kohler S, Satyanarayana G, Vandunk C et al (2006) Selected neuropeptides, and cytokines: developmental trajectory and concentrations in neonatal blood of children with autism or Down syndrome. Int J Dev Neurosci 241:73–80CrossRefGoogle Scholar
  42. 42.
    Perry EK, Lee MW, Martin-Ruiz CM, Court JA, Volsen SG, Merrit J et al (2001) Cholinergic activity in autism: abnormalities in the cerebral cortex and basal forebrain. Am J Psychiatry 158:1058–1066PubMedCrossRefGoogle Scholar
  43. 43.
    Ghosh A, Greenberg ME (1995) Distinct roles for bFGF and NT-3 in the regulation of cortical neurogenesis. Neuron 15:89–103PubMedCrossRefGoogle Scholar
  44. 44.
    Segal RA, Pomeroy SL, Stiles CD (1995) Axonal growth and fasciculation linked to differential expression of BDNF and NT-3 receptors in developing granule cells. J Neurosci 15:4970–4981PubMedGoogle Scholar
  45. 45.
    Bates B, Hirt L, Thomas SS, Akbarian S, Le D, Amin-Hanjani S et al (2002) Neurotrophin-3 promotes cell death induced in cerebellar ischemia, oxygen-glucose deprivation, and oxidative stress: possible involvement of oxygen free radicals. Neurobiol Dis 9:24–37PubMedCrossRefGoogle Scholar
  46. 46.
    Pasarica D, Gheorghiu M, Toparceanu F, Bleotu C, Ichim L, Trandafir T (2005) Neurotrophin-3, TNF-alpha and IL-6 relations in serum and cerebrospinal fluid of ischemic stroke patients. Roum Arch Microbiol Immunol 64:27–33PubMedGoogle Scholar
  47. 47.
    Morrison ME, Mason CA (1998) Granule neuron regulation of Purkinje cell development: striking a balance between neurotrophin and glutamate signaling. J Neurosci 18:3563–3573PubMedGoogle Scholar
  48. 48.
    Cappelletti G, Maggioni MG, Tedeschi G, Maci R (2003) Protein tyrosine nitration is triggered by nerve growth factor during neuronal differentiation of PC12 cells. Exp Cell Res 288:9–20PubMedCrossRefGoogle Scholar
  49. 49.
    Vargas MR, Pehar M, Cassina P, Estavez AG, Beckman JS, Barbeito L (2004) Stimulation of nerve growth factor expression in astrocytes by peroxynitrite. In vivo 18:269–274PubMedGoogle Scholar
  50. 50.
    Behrens MM, Strasser U, Lobner D, Dugan LL (1999) Neurotrophin-mediated potentiation of neuronal injury. Microsc Res Tech 45:276–284PubMedCrossRefGoogle Scholar
  51. 51.
    Yip J, Soghomonian JJ, Blatt GJ (2007) Decreased GAD67 mRNA levels in cerebellar Purkinje cells in autism: pathophysiological implications. Acta Neuropathol 113:559–568PubMedCrossRefGoogle Scholar
  52. 52.
    Rout UK, Dhossche DM (2008) A pathogenetic model of autism involving Purkinje cell loss through anti GAD antibodies. Med Hypotheses 71:218–221PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2009

Authors and Affiliations

  • Elizabeth M. Sajdel-Sulkowska
    • 1
    • 2
  • Ming Xu
    • 3
  • Noriyuki Koibuchi
    • 3
  1. 1.Department of PsychiatryHarvard Medical SchoolBostonUSA
  2. 2.Department of PsychiatryBrigham and Women’s HospitalBostonUSA
  3. 3.Department of Integrative Physiology, Graduate School of MedicineGunma UniversityMaebashiJapan

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