Advertisement

The survival response of mesencephalic dopaminergic neurons to the neurotrophins BDNF and NT-4 requires priming with serum: comparison with members of the TGF-β superfamily and characterization of the serum-free culture system

  • K. Krieglstein
  • D. Maysinger
  • K. Unsicker
Part of the Journal of Neural Transmission Supplement book series (NEURAL SUPPL, volume 47)

Summary

The neurotrophins, brain-derived neurotrophic factor (BDNF) and neurotrophin-4 (NT-4), are established survival promoting molecules for dopaminergic (DAergic) neurons cultured from the fetal rat midbrain floor. We have cultured and compared the survival of embryonic day (E) 14 mesencephalic cells in fully defined, serum-free medium, with serum-primed cultures (one hour during dissociation). Cultures were characterized using antibodies against neuron-specific enolase (NSE), tyrosine hydroxylase (TH), vimentin, glial fibrillary acidic protein (GFAP), and the antigen A2B5. The absolute absence of serum did not reduce the survival of TH-positive DAergic neurons nor alter the percentages of cells staining for the above markers. Transforming growth factor-β3 (TGF-β3) and glial cell line-derived neurotrophic factor (GDNF), two members of the TGF-β superfamily, both promoted the survival of TH-positive cells (TGF-β3: 2-fold; GDNF: 1.6-fold) over the 8-day culture period. Survival mediated by TGF-β3 and GDNF was independent of whether or not the cells had been initially exposed to serum. In contrast, the survival promoting effects of BDNF and NT-4 were crucially dependent on serum priming. RT-PCR for the full-length trkB high affinity neurotrophin receptor revealed its presence in both culture systems. We conclude that priming with serum is important to make DAergic neurons fully responsive to BDNF and NT-4. Underlying mechanisms might be sought at the level or distal of trkB receptor expression, without excluding the possiblity that serum elicits production of growth factors that synergistically act with neurotrophins in these cultures.

Keywords

Glial Fibrillary Acidic Protein Tyrosine Hydroxylase Dopaminergic Neuron DAergic Neuron Midbrain Dopaminergic Neuron 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Beck KD (1994) Functions of brain-derived neurotrophic factor, insulin-like growth factor-1 and basic fibroblast growth factor in the development and maintenance of dopaminergic neurons. Prog Neurobiol 44: 497–451PubMedCrossRefGoogle Scholar
  2. Beck KD, Knüsel 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: 855–866PubMedCrossRefGoogle Scholar
  3. Beck KD, Valverde J, Alexi T, Poulsen K, Moffat B, Vandlen RA, Rosenthal A, Hefti F (1995) Mesencephalic dopaminergic neurons protected by GDNF from axotomyinduced degeneration in the adult brain. Nature 373: 339–341PubMedCrossRefGoogle Scholar
  4. Bothwell M (1995) Functional interactions of neurotrophins and neurotrophin receptors. Annu Rev Neurosci 18: 223–253PubMedCrossRefGoogle Scholar
  5. Cohen G, Werner P (1994) Free radicals, oxidative stress, and neurodegeneration. In: Calne DB (ed) Neurodegenerative diseases. Saunders, pp 139–161Google Scholar
  6. Edwards RH (1993) Pathogenesis of Parkinson’s disease. Clin Neurosci 1: 36–44Google Scholar
  7. Engele J, Bohn MC (1991) The neurotrophic effects of fibroblast growth factors on dopaminergic neurons in vitro are mediated by mesencephalic glia. J Neurosci 11: 3070–3078PubMedGoogle Scholar
  8. Gerlach M, Ben-Shachar D, Riederer P, Youdim MBH (1994) Altered brain metabolism of iron as a cause of neurodegenerative diseases? J Neurochem 63: 793–807PubMedCrossRefGoogle Scholar
  9. Götz ME, Künig G, Riederer P, Youdim MBH (1994) Oxidative stress: free radical production in neural degeneration. Pharmacol Ther 63: 37–122PubMedCrossRefGoogle Scholar
  10. Hantzopoulos PA, Suri C, Glass DJ, Goldfarb MP, Yancopoulos GD (1994) The low affinity NGF receptor, p75, can collaborate with each of the trks to potentiate functional responses to the neurotrophins Neuron 13: 187–201Google Scholar
  11. Hoffer BJ, Hoffman A, Bowenkamp K, Huettl P, Hudson J, Martin D, Lin L-FH, Gerhardt GA (1994) Glial cell line-derived neurotrophic factor reverses toxin-induced injury to midbrain dopaminergic neurons in vivo. Neurosci Lett 182: 107–111PubMedCrossRefGoogle Scholar
  12. Hyman C, Hoffer M, Barde Y-A, Juhasz M, Yancopoulos GD, Squinto SP, Lindsay RM (1991) BDNF is a neurotrophic factor for dopaminergic neurons of the substantia nigra. Nature 350: 230–232.PubMedCrossRefGoogle Scholar
  13. Hyman C, Juhasz M, Jackson C, Wright P, Ip NY, Lindsay RM (1993) Overlapping and distinct actions of the neurotrophins, BDNF, NT-3 and NT-4/5, on cultured dopaminergic and GABAegic neurons of the ventral mesencephalon. J Neurosci 14: 335–347Google Scholar
  14. Hynes MA, Poulson K, Armanini M, Berkemeier L, Phillips H, Rosenthal A (1994) Neurotrophin-4/5 is a survival factor for embryonic midbrain dopaminergic neurons in enriched cultures. J Neurosci Res 37: 144–154PubMedCrossRefGoogle Scholar
  15. Jackson GR, Apffel L, Werrbach-Perez K, Perez-Polo JR (1990) Role of nerve growth factor in oxidant-antioxidant balance and neuronal injury. I. Stimulation of hydrogen peroxide resistance. J Neurosci Res 25: 360–368PubMedCrossRefGoogle Scholar
  16. Kaplan DR, Matsumoto K, Lucarelli E, Thiele CJ (1993) Induction of TrkB by retinoid acid mediates biologic responsiveness to BDNF and differentiation of human neuro-blastoma cells. Neuron 11: 321–331PubMedCrossRefGoogle Scholar
  17. Krieglstein K, Unsicker K (1994) TGF-(3 promotes survival of midbrain dopmainergic neurons and protects them against MPP+ toxicity. Neuroscience 63: 1189–1196PubMedCrossRefGoogle Scholar
  18. Krieglstein K, Suter-Crazzolara C, Fischer WH, Unsicker K (1995) TGF-(3 superfamily members promote survival of midbrain dopaminergic neurons and protect them against MPP+ toxicity. EMBO J 14: 736–742PubMedGoogle Scholar
  19. Levi-Montalcini R (1987) The nerve growth factor 35 years later. Science 237: 1154–1162PubMedCrossRefGoogle Scholar
  20. Lin L-FH, Doherty DH, Lile JD, Bektesh S, Collins F (1993) GDNF: a glial cell line-derived neurotrophic factor for midbrain dopaminergic neurons. Science 246:1023–1025CrossRefGoogle Scholar
  21. Lindsay RM, Altar CA, Cedarbaum JM, Hyman C, Wiegand SJ (1993) The therapeutic potential of neurotrophic factors in the treatment of Parkinson’s disease. Exp Neurol 124: 103–118PubMedCrossRefGoogle Scholar
  22. Malarkey K, Belham CM, Paul A, Graham A, McLees A, Scott PH, Plevin R (1995) The regulation of tyrosine kinase signalling pathways by growth factor and G-coupled receptors. Biochem J 309: 361–375PubMedGoogle Scholar
  23. Mattson MP, Scheff SW (1994) Endogenous neuroprotection factors and traumatic brain injury: mechanisms of action and implications for therapy. J Neurotrauma 11: 3–33PubMedCrossRefGoogle Scholar
  24. Middlemas DS, Lindberg RA, Hunter T (1991) TrkB, a neural receptor protein-tyrosine kinase: evidence for full-length and two truncated receptors. GenBank M55291Google Scholar
  25. Mufson EJ, Kroin JS, Sobreviela T, Burke MA, Kordower JH, Penn RD, Miller JA (1994) Intrastriatal infusions of brain-derived neurotrophic factor: retrograde transport and colocalization with dopamine containing substantia nigra neurons in rat. Exp Neurol 129: 15–26PubMedCrossRefGoogle Scholar
  26. Poulsen KT, Armanini MP, Klein RD, Hynes MA, Phillips HS, Rosenthal A (1994) TGF132 and TGF133 are potent survival factors for midbrain dopaminergic neurons. Neuron 13: 1245–1252PubMedCrossRefGoogle Scholar
  27. Reichmann H, Riederer P (1994) Mitochondrial disturbances in neurodegeneration. In: Calne DB (ed) Neurodegenerative diseases. Saunders, Philadelphia, pp 195–204Google Scholar
  28. Schapira AHV (1995) Oxidative stress in Parkinson’s disease. Neuropathol ApplGoogle Scholar
  29. Neurobiol 21: 3–9Google Scholar
  30. Shimoda K, Sauve Y, Marini A, Schwartz JP, Commissiong JW (1992) A high percentage of tyrosine hydroxylase-positive cells from rat E14 mesencephalic cell culture. Brain Res 586: 319–331PubMedCrossRefGoogle Scholar
  31. Takeshima T, Johnston JM, Commissiong JW (1994) Mesencephalic type 1 astrocytes rescue dopaminergic neurons from death induced by serum deprevation. J Neurosci 14: 4769–4779PubMedGoogle Scholar
  32. Tomac A, Lindqvist E, Lin L-FH, Imgren SO, Young D, Hoffer BJ, Olson L (1995) Protection and repair of the nigrostriatal dopaminergic system by GDNF in vivo. Nature 373: 335–339PubMedCrossRefGoogle Scholar
  33. Unsicker K (1994) Growth factors in Parkinson’s disease. Prog Growth Factor Res 5: 73–87PubMedCrossRefGoogle Scholar
  34. Zhang Y, Tatsuno T, Carney JM, Mattson MP (1993) Basic FGF, NGF, and IGFs protect hippocampal and cortical neurons against iron-induced degeneration. J Cereb Blood Flow Metab 13: 378–388PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag 1996

Authors and Affiliations

  • K. Krieglstein
    • 1
  • D. Maysinger
    • 2
  • K. Unsicker
    • 1
  1. 1.Department of Anatomy and Cell BiologyUniversity of HeidelbergHeidelbergFederal Republic of Germany
  2. 2.PharmacologyMcGill UniversityMontrealCanada

Personalised recommendations