Experimental Brain Research

, Volume 234, Issue 7, pp 1863–1873 | Cite as

IGF-1 protects dopamine neurons against oxidative stress: association with changes in phosphokinases

  • Amina El Ayadi
  • Michael J. Zigmond
  • Amanda D. SmithEmail author
Research Article


Insulin-like growth factor-1 (IGF-1) is an endogenous peptide transported across the blood brain barrier that is protective in several brain injury models, including an acute animal model of Parkinson’s disease (PD). Motor deficits in PD are due largely to the progressive loss of nigrostriatal dopaminergic neurons. Thus, we examined the neuroprotective potential of IGF-1 in a progressive model of dopamine deficiency in which 6-hydroxydopamine (6-OHDA) is infused into the striatum. Rats received intrastriatal IGF-1 (5 or 50 µg) 6 h prior to infusion of 4 µg 6-OHDA into the same site and were euthanized 1 or 4 weeks later. Both concentrations of IGF-1 protected tyrosine hydroxylase (TH) immunoreactive terminals in striatum at 4 weeks but not at 1 week, indicating that IGF-induced restoration of the dopaminergic phenotype occurred over several weeks. TH-immunoreactive cell loss was only attenuated with 50 µg IGF-1. We then examined the effect of striatal IGF-1 on the Ras/ERK1/2 and PI3K/Akt pathways to ascertain whether their activation correlated with IGF-1-induced protection. Striatal and nigral levels of phospho-ERK1/2 were maximal 6 h after IGF-1 infusion and, with the exception of an increase in nigral pERK2 at 48 h, returned to basal levels by 7 days. Phospho-Akt (Ser473) was elevated 6-24 h post-IGF-1 infusion in both striatum and substantia nigra concomitant with inhibition of pro-death GSK-3β, a downstream target of Akt. These results suggest that IGF-1 can protect the nigrostriatal pathway in a progressive PD model and that this protection is preceded by activation of key pro-survival signaling cascades.


6-OHDA Akt CREB Parkinson’s disease Striatum ERK1/2 GSK-3Beta 



The study was funded by Career Development Award (NS45698) to ADS, Rita Levi-Montalcini Fellowship to AE and both a Udall Center of Excellence in Parkinson’s Disease Research (NS19608) and a Research Project Grant Award (NS070825) to MJZ.


  1. Abe M, Kimoto H, Eto R, Sasaki T, Kato H, Kasahara J, Araki T (2010) Postnatal development of neurons, interneurons and glial cells in the substantia nigra of mice. Cell Mol Neurobiol 30:917–928CrossRefPubMedGoogle Scholar
  2. Aberg ND, Brywe KG, Isgaard J (2006) Aspects of growth hormone and insulin-like growth factor-I related to neuroprotection, regeneration, and functional plasticity in the adult brain. Sci World J 6:53–80CrossRefGoogle Scholar
  3. Aberle H, Bauer A, Stappert J, Kispert A, Kemler R (1997) Beta-catenin is a target for the ubiquitin–proteasome pathway. EMBO J 16:3797–3804CrossRefPubMedPubMedCentralGoogle Scholar
  4. Agid Y, Blin J (1987) Nerve cell death in degenerative disease of the central nervous system: clinical aspects. Ciba Found Symph 126:3–29Google Scholar
  5. Airavaara M, Voutilainen MH, Wang Y, Hoffer B (2012) Neurorestoration. Parkinsonism Relat Disord 18:S143–S146CrossRefPubMedPubMedCentralGoogle Scholar
  6. Alonso M, Vianna MR, Izquierdo I, Medina JH (2002) Signaling mechanisms mediating BDNF modulation of memory formation in vivo in the hippocampus. Cell Mol Neurobiol 22:663–674CrossRefPubMedGoogle Scholar
  7. Beck KD, Knusel B, Hefti F (1993) The nature of the trophic action of brain-derived neurotrophic factor, des(1-3)-insulin-like growth factor-1, and basic fibroblast growth factor on mesencephalic dopaminergic neurons developing in culture. Neuroscience 52:855–866CrossRefPubMedGoogle Scholar
  8. Boger HA, Middaugh LD, Huang P, Zaman V, Smith AC, Hoffer BJ, Tomac AC, Granholm AC (2006) A partial GDNF depletion leads to earlier age-related deterioration of motor function and tyrosine hydroxylase expression in the substantia nigra. Exp Neurol 202:336–347CrossRefPubMedGoogle Scholar
  9. Bowenkamp KE, David D, Lapchak PL, Henry MA, Granholm AC, Hoffer BJ, Mahalik TJ (1996) 6-hydroxydopamine induces the loss of the dopaminergic phenotype in substantia nigra neurons of the rat. A possible mechanism for restoration of the nigrostriatal circuit mediated by glial cell line-derived neurotrophic factor. Exp Brain Res 111:1–7CrossRefPubMedGoogle Scholar
  10. Burke RE, O’Malley K (2013) Axon degeneration in Parkinson’s disease. Exp Neurol 246:72–83CrossRefPubMedPubMedCentralGoogle Scholar
  11. Calabrese F, Guidotti G, Racagni G, Riva MA (2013) Reduced neuroplasticity in aged rats: a role for the neurotrophin brain-derived neurotrophic factor. Neurobiol Aging 34:2768–2776CrossRefPubMedGoogle Scholar
  12. Cardona-Gomez GP, Mendez P, Don Carlos LL, Azcoitia I, Garcia-Segura LM (2002) Interactions of estrogen and insulin-like growth factor-I in the brain: molecular mechanisms and functional implications. J Steroid Biochem Mol Biol 83:211–217CrossRefPubMedGoogle Scholar
  13. Carro E, Trejo JL, Busiguina S, Torres-Aleman I (2001) Circulating insulin-like growth factor I mediates the protective effects of physical exercise against brain insults of different etiology and anatomy. J Neurosci 21:5678–5684PubMedGoogle Scholar
  14. Chauhan NB, Siegel GJ, Lee JM (2001) Depletion of glial cell line-derived neurotrophic factor in substantia nigra neurons of Parkinson’s disease brain. J Chem Neuroanat 21:277–288CrossRefPubMedGoogle Scholar
  15. Cohen AD, Zigmond MJ, Smith AD (2011) Effects of intrastriatal GDNF on the response of dopamine neurons to 6-hydroxydopamine: time course of protection and neurorestoration. Brain Res 1370:80–88CrossRefPubMedPubMedCentralGoogle Scholar
  16. De Keyser J, Wilczak N, De Backer JP, Herroelen L, Vauquelin G (1994) Insulin-like growth factor-I receptors in human brain and pituitary gland: an autoradiographic study. Synapse 17:196–202CrossRefPubMedGoogle Scholar
  17. Foncea R, Andersson M, Ketterman A, Blakesley V, Sapag-Hagar M, Sugden PH, LeRoith D, Lavandero S (1997) Insulin-like growth factor-I rapidly activates multiple signal transduction pathways in cultured rat cardiac myocytes. J Biol Chem 272:19115–19124CrossRefPubMedGoogle Scholar
  18. Georgievska B, Kirik D, Björklund A (2002) Aberrant sprouting and downregulation of tyrosine hydroxylase in lesioned nigrostriatal dopamine neurons induced by long-lasting overexpression of glial cell line derived neurotrophic factor in the striatum by lentiviral gene transfer. Exp Neurol 177:461–474CrossRefPubMedGoogle Scholar
  19. German DC, Manaye KF (1993) Midbrain dopaminergic neurons (nuclei A8, A9, and A10): three-dimensional reconstruction in the rat. J Comp Neurol 331:297–309CrossRefPubMedGoogle Scholar
  20. Gill SS, Patel NK, Hotton GR, O’Sullivan K, McCarter R, Bunnage M, Brooks DJ, Svendsen CN, Heywood P (2003) Direct brain infusion of glial cell line-derived neurotrophic factor in Parkinson disease. Nat Med 9:589–595CrossRefPubMedGoogle Scholar
  21. Guan J, Krishnamurthi R, Waldvogel HJ, Faull RLM, Clark R, Gluckman P (2000) N-terminal tripeptide of IGF-1 (GPE) prevents the loss of TH positive neurons after 6-OHDA induced nigral lesion in rats. Brain Res 859:286–292CrossRefPubMedGoogle Scholar
  22. Hornykeiwicz O (1998) Biochemical aspects of Parkinson’s disease. Neurology 51:S2–S9CrossRefGoogle Scholar
  23. Howells DW, Porritt MJ, Wong JY, Batchelor PE, Kalnins R, Hughes AJ, Donnan GA (2000) Reduced BDNF mRNA expression in the Parkinson’s disease substantia nigra. Exp Neurol 166:127–135CrossRefPubMedGoogle Scholar
  24. Hwang YP, Kim HG, Han EH, Jeong HG (2008) Metallothionein-III protects against 6-hydroxydopamine-induced oxidative stress by increasing expression of heme oxygenase-1 in a PI3K and ERK/Nrf2-dependent manner. Toxicol Appl Pharmacol 231:318–327CrossRefPubMedGoogle Scholar
  25. Kim KC, Kang KA, Zhang R, Piao MJ, Kim GY, Kang MY, Lee SJ, Lee NH, Surh YJ, Hyun JW (2010) Up-regulation of Nrf2-mediated heme oxygenase-1 expression by eckol, a phlorotannin compound, through activation of Erk and PI3K/Akt. Int J Biochem Cell Biol 42(2):297–305CrossRefPubMedGoogle Scholar
  26. Koopmans GC, Brans M, Gomez-Pinilla F, Duis S, Gispen WH, Torres-Aleman I, Joosten EA, Hamers FP (2006) Circulating insulin-like growth factor I and functional recovery from spinal cord injury under enriched housing conditions. Eur J Neurosci 23:1035–1046CrossRefPubMedGoogle Scholar
  27. Krishnamurthi R, Stott S, Maingay M, Faull RL, McCarthy D, Gluckman P, Guan J (2004) N-terminal tripeptide of IGF-1 improves functional deficits after 6-OHDA lesion in rats. NeuroReport 15:1601–1604CrossRefPubMedGoogle Scholar
  28. Kulich SM, Chu CT (2001) Sustained extracellular signal-regulated kinase activation by 6-hydroxydopamine: implications for Parkinson’s disease. J Neurochem 77:1058–1066CrossRefPubMedPubMedCentralGoogle Scholar
  29. Leinninger GM, Backus C, Uhler MD, Lentz SI, Feldman EL (2004) Phosphatidylinositol 3-kinase and Akt effectors mediate insulin-like growth factor-I neuroprotection in dorsal root ganglia neurons. FASEB J 18:1544–1546PubMedGoogle Scholar
  30. LeRoith D, Werner H, Faria TN, Kato H, Adamo M, Roberts CT Jr (1993) Insulin-like growth factor receptors. Implications for nervous system function. Ann N Y Acad Sci 692:22–32CrossRefPubMedGoogle Scholar
  31. Love S, Plaha P, Patel NK, Hotton GR, Brooks DJ, Gill SS (2005) Glial cell line-derived neurotrophic factor induces neuronal sprouting in human brain. Nat Med 11:703–704CrossRefPubMedGoogle Scholar
  32. Lu X, Hagg T (1997) Glial cell line-derived neurotrophic factor prevents death, but not reductions in tyrosine hydroxylase, of injured nigrostriatal neurons in adult rats. J Comp Neurol 388:484–494CrossRefPubMedGoogle Scholar
  33. Maggio M, Ble A, Ceda GP, Metter EJ (2006) Decline in insulin-like growth factor-I levels across adult life span in two large population studies. J Gerontol A Biol Sci Med Sci 61:182–183CrossRefPubMedGoogle Scholar
  34. Markuns JF, Wojtaszewski JF, Goodyear LJ (1999) Insulin and exercise decrease glycogen synthase kinase-3 activity by different mechanisms in rat skeletal muscle. J Biol Chem 274:24896–24900CrossRefPubMedGoogle Scholar
  35. Nair-Roberts RG, Chatelain-Badie SD, Benson E, White-Cooper H, Bolam JP, Ungless MA (2008) Stereological estimates of dopaminergic, GABAergic and glutamatergic neurons in the ventral tegmental area, substantia nigra and retrorubral field in the rat. Neuroscience 152:1024–1031CrossRefPubMedPubMedCentralGoogle Scholar
  36. Nutt JG, Burchiel KJ, Comella CL, Jankovic J, Lang AE, Laws ER Jr, Lozano AM, Penn RD, Simpson RK Jr, Stacy M, Wooten GF (2003) Randomized, double-blind trial of glial cell line-derived neurotrophic factor (GDNF) in PD. Neurology 60:69–73CrossRefPubMedGoogle Scholar
  37. O’Callaghan JP, Sriram K (2004) Focused microwave irradiation of the brain preserves in vivo protein phosphorylation: comparison with other methods of sacrifice and analysis of multiple phosphoproteins. J Neurosci Methods 135:159–168CrossRefPubMedGoogle Scholar
  38. Ortega F, Perez-Sen R, Delicado EG, Miras-Portugal MT (2011) ERK1/2 activation is involved in the neuroprotective action of P2Y13 and P2X7 receptors against glutamate excitotoxicity in cerebellar granule neurons. Neuropharmacology 61:1210–1221CrossRefPubMedGoogle Scholar
  39. Pan W, Kastin AJ (2000) Interactions of IGF-1 with the blood brain barrier in vivo and in situ. Neuroendocrinology 72:171–178CrossRefPubMedGoogle Scholar
  40. Parain K, Murer MG, Yan Q, Faucheux B, Agid Y, Hirsch E, Raisman-Vozari R (1999) Reduced expression of brain-derived neurotrophic factor protein in Parkinson’s disease substantia nigra. NeuroReport 10:557–561CrossRefPubMedGoogle Scholar
  41. Patel NK, Pavese N, Javed S, Hotton GR, Brooks DJ, Gill SS (2013) Benefits of putaminal GDNF infusion in Parkinson disease are maintained after GDNF cessation. Neurology 81:1176–1178CrossRefPubMedPubMedCentralGoogle Scholar
  42. Paxinos G, Watson C (1982) The Rat Brain in Stereotaxic Coordinates. Academic Press, SydneyGoogle Scholar
  43. Quesada A, Micevych PE (2004) Estrogen interacts with the IGF-1 system to protect nigrostriatal dopamine and maintain motoric behavior after 6-hydroxdopamine lesions. J Neurosci Res 75:107–116CrossRefPubMedGoogle Scholar
  44. Quesada A, Romeo HE, Micevych P (2007) Distribution and localization patterns of estrogen receptor-beta and insulin-like growth factor-1 receptors in neurons and glial cells of the female rat substantia nigra: localization of ERbeta and IGF-1R in substantia nigra. J Comp Neurol 503:198–208CrossRefPubMedPubMedCentralGoogle Scholar
  45. Quesada A, Lee BY, Micevych PE (2008) PI3 kinase/Akt activation mediates estrogen and IGF-1 nigral DA neuronal neuroprotection against a unilateral rat model of Parkinson’s disease. Dev Neurobiol 68:632–644CrossRefPubMedPubMedCentralGoogle Scholar
  46. Rosenblad C, Kirik D, Devaux B, Moffat B, Phillips HS, Bjorklund A (1999) Protection and regeneration of nigral dopaminergic neurons by neurturin or GDNF in a partial lesion model of Parkinson’s disease after administration into the striatum or the lateral ventricle. Eur J Neurosci 11:1554–1566CrossRefPubMedGoogle Scholar
  47. Rotwein P, Burgess SK, Milbrandt JD, Krause JE (1988) Differential expression of insulin-like growth factor genes in rat central nervous system. Proc Nat Acad Sci 85:265–269CrossRefPubMedPubMedCentralGoogle Scholar
  48. Sauer H, Oertel WH (1994) Progressive degeneration of nigrostriatal dopamine neurons following intrastriatal terminal lesions with 6-hydroxydopamine: a combined retrograde tracing and immunocytochemical study in the rat. Neuroscience 59:401–415CrossRefPubMedGoogle Scholar
  49. Sherer TB, Fiske BK, Svendsen CN, Lang AE, Langston JW (2006) Crossroads in GDNF therapy for Parkinson’s disease. Mov Disord 21:136–141CrossRefPubMedGoogle Scholar
  50. Slevin JT, Gash DM, Smith CD, Gerhardt GA, Kryscio R, Chebrolu H, Walton A, Wagner R, Young AB (2006) Unilateral intraputaminal glial cell line-derived neurotrophic factor in patients with Parkinson disease: response to 1 year each of treatment and withdrawal. Neurosurg Focus 20:E1CrossRefPubMedGoogle Scholar
  51. Sonntag WE, Lynch CD, Bennett SA, Khan AS, Thornton PL, Cooney PT, Ingram RL, McShane T, Brunso-Bechtold JK (1999) Alterations in insulin-like growth factor-1 gene and protein expression and type 1 insulin-like growth factor receptors in the brains of ageing rats. Neuroscience 88:269–279CrossRefPubMedGoogle Scholar
  52. Stanciu M, Wang Y, Kentor R, Burke N, Watkins S, Kress G, Reynolds I, Klann E, Angiolieri MR, Johnson JW, DeFranco DB (2000) Persistent activation of ERK contributes to glutamate-induced oxidative toxicity in a neuronal cell line and primary cortical neuron cultures. J Biol Chem 275:12200–12206CrossRefPubMedGoogle Scholar
  53. Subramaniam S, Zirrgiebel U, Und Halbach OVB, Strelau J, Laliberte C, Kaplan DR, Unsicker K (2004) ERK activation promotes neuronal degeneration predominantly through plasma membrane damage and independently of caspase-3. J Cell Biol 165:357–369CrossRefPubMedPubMedCentralGoogle Scholar
  54. Trejo JL, Carro E, Torres-Aleman I (2001) Circulating insulin-like growth factor I mediates exercise-induced increases in the number of new neurons in the adult hippocampus. J Neurosci 21:1628–1634PubMedGoogle Scholar
  55. Willaime-Morawek S, Arbez N, Mariani J, Brugg B (2005) IGF-I protects cortical neurons against ceramide-induced apoptosis via activation of the PI-3 K/Akt and ERK pathways; is this protection independent of CREB and Bcl-2? Mol Brain Res 142:97–106CrossRefPubMedGoogle Scholar
  56. Yaghmaie F, Saeed O, Garan SA, Voelker MA, Gouw AM, Freitag W, Sternberg H, Timiras PS (2006) Age-dependent loss of insulin-like growth factor-1 receptor immunoreactive cells in the supraoptic hypothalamus is reduced in calorically restricted mice. Int J Dev Neurosci 24:431–436CrossRefPubMedGoogle Scholar
  57. Yurek DM, Fletcher-Turner A (2001) Differential expression of GDNF, BDNF, and NT-3 in the aging nigrostriatal system following a neurotoxic lesion. Brain Res 89:228–235CrossRefGoogle Scholar
  58. Zawada WM, Kirschman DL, Cohen JJ, Heidenreich KA, Freed CR (1996) Growth factors rescue embryonic dopamine neurons from programmed cell death. Exp Neurol 140:60–67CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2016

Authors and Affiliations

  • Amina El Ayadi
    • 1
    • 3
  • Michael J. Zigmond
    • 1
  • Amanda D. Smith
    • 1
    • 2
    Email author
  1. 1.Pittsburgh Institute for Neurodegenerative DiseaseUniversity of PittsburghPittsburghUSA
  2. 2.VA Pittsburgh Healthcare SystemUniversity of PittsburghPittsburghUSA
  3. 3.Department of SurgeryUniversity of Texas Medical BranchGalvestonUSA

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