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.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Preview
Unable to display preview. Download preview PDF.
References
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: 497–451
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: 855–866
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: 339–341
Bothwell M (1995) Functional interactions of neurotrophins and neurotrophin receptors. Annu Rev Neurosci 18: 223–253
Cohen G, Werner P (1994) Free radicals, oxidative stress, and neurodegeneration. In: Calne DB (ed) Neurodegenerative diseases. Saunders, pp 139–161
Edwards RH (1993) Pathogenesis of Parkinson’s disease. Clin Neurosci 1: 36–44
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: 3070–3078
Gerlach M, Ben-Shachar D, Riederer P, Youdim MBH (1994) Altered brain metabolism of iron as a cause of neurodegenerative diseases? J Neurochem 63: 793–807
Götz ME, Künig G, Riederer P, Youdim MBH (1994) Oxidative stress: free radical production in neural degeneration. Pharmacol Ther 63: 37–122
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: 187–201
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: 107–111
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.
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: 335–347
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: 144–154
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: 360–368
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: 321–331
Krieglstein K, Unsicker K (1994) TGF-(3 promotes survival of midbrain dopmainergic neurons and protects them against MPP+ toxicity. Neuroscience 63: 1189–1196
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: 736–742
Levi-Montalcini R (1987) The nerve growth factor 35 years later. Science 237: 1154–1162
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 246:1023–1025
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: 103–118
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: 361–375
Mattson MP, Scheff SW (1994) Endogenous neuroprotection factors and traumatic brain injury: mechanisms of action and implications for therapy. J Neurotrauma 11: 3–33
Middlemas DS, Lindberg RA, Hunter T (1991) TrkB, a neural receptor protein-tyrosine kinase: evidence for full-length and two truncated receptors. GenBank M55291
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: 15–26
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: 1245–1252
Reichmann H, Riederer P (1994) Mitochondrial disturbances in neurodegeneration. In: Calne DB (ed) Neurodegenerative diseases. Saunders, Philadelphia, pp 195–204
Schapira AHV (1995) Oxidative stress in Parkinson’s disease. Neuropathol Appl
Neurobiol 21: 3–9
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: 319–331
Takeshima T, Johnston JM, Commissiong JW (1994) Mesencephalic type 1 astrocytes rescue dopaminergic neurons from death induced by serum deprevation. J Neurosci 14: 4769–4779
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: 335–339
Unsicker K (1994) Growth factors in Parkinson’s disease. Prog Growth Factor Res 5: 73–87
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: 378–388
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 1996 Springer-Verlag
About this paper
Cite this paper
Krieglstein, K., Maysinger, D., Unsicker, K. (1996). 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. In: Jellinger, K.A., Windisch, M. (eds) New Trends in the Diagnosis and Therapy of Non-Alzheimer’s Dementia. Journal of Neural Transmission Supplement, vol 47. Springer, Vienna. https://doi.org/10.1007/978-3-7091-6892-9_17
Download citation
DOI: https://doi.org/10.1007/978-3-7091-6892-9_17
Publisher Name: Springer, Vienna
Print ISBN: 978-3-211-82823-6
Online ISBN: 978-3-7091-6892-9
eBook Packages: Springer Book Archive