Abstract
Dopaminergic neuronal cell degeneration is the principal characteristic feature of the neuropathology of Parkinson’s disease. Cultures of mesencephalic neurons are widely used as a source of dopaminergic neurons for the study of mechanisms implicated in dopaminergic cell death and for the evaluation of potential dopaminergic neuroprotective agents, including neurotrophic factors. This chapter presents a detailed protocol for the preparation of rat mesencephalic cell cultures and their application to evaluating the neuroprotective action of brain-derived neurotrophic factor.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Forno LS (1996) Neuropathology of Parkinson’s disease. J Neuropathol Exp Neurol 55, 259–272
Gelb DJ, Oliver E, and Gilman S (1999) Diagnostic criteria for Parkinson disease. Arch Neurol 56, 33–39
Hyman C, Hofer M, Barde Y-A, Juhasz M, Yancopoulos GD, Squinto SP, et al. (1991) BDNF is a neurotrophic factor for dopaminergic neurons in the substantia nigra. Nature 350, 230–232
Knüsel B, Winslow JW, Rosenthal A, Burton LE, Seid DP, Nikolics K, et al (1991) Promotion of central cholinergic and dopaminergic neuron differentiation by brain-derived neurotrophic factor but not neurotrophin-3. Proc Natl Acad Sci USA 88, 961–965
Somoza R, Juri C, Baes M, Wyneken U, and Rubio FJ (2010) Intranigral transplantation of epigenetically-induced BDNF-secreting human mesenchymal stem cells: implications for cell-based therapies in Parkinson s disease. Biol Blood Marrow Transplant 16, 1530–1540
Wilkins A, Kemp K, Ginty M, Hares K, Mallam E, and Scolding N. Human bone marrow-derived mesenchymal stem cells secrete brain-derived neurotrophic factor which promotes neuronal survival in vitro. Stem Cell Res 2009 Mar 27. (Epub ahead of print)
Wang R, Li YH, Xu Y, Li YB, Wu HL, Guo H, et al (2010) Curcumin produces neuroprotective effects via activating brain-derived neurotrophic factor/TrkB-dependent MAPK and PI-3K cascades in rodent cortical neurons. Prog Neuropsychopharmacol Biol Psychiatry 34, 147–153
Wu CL, Chen SD, Yin JH, Hwang CS, and Yang DI (2010) Erythropoietin and sonic hedgehog mediate the neuroprotective effects of brain-derived neurotrophic factor against mitochondrial inhibition. Neurobiol Dis 40, 146–154
Prochiantz A, Di Porzio U, Kato A, Berger B, and Glowinski J (1979) In vitro maturation of mesencephalic dopaminergic neurons from mouse embryos is enhanced in presence of their striatal target cells. Proc Natl Acad Sci USA 76, 5387–5391
Ferrari G, Minozzi M-C, Toffano G, Leon A, and Skaper SD (1989) Basic fibroblast growth factor promotes the survival and development of mesencephalic neurons in culture. Dev Biol 133, 140–147
Qin L, Liu Y, Wang T, Wei SJ, Block ML, Wilson B, et al (2004) NADPH oxidase mediates lipopolysaccharide-induced neurotoxicity and proinflammatory gene expression in activated microglia. J Biol Chem 279, 1415–1421
Gao HM, Liu B, and Hong JS (2003) Critical role for microglial NADPH oxidase in rotenone-induced degeneration of dopaminergic neurons. J Neurosci 23, 6181–6187
Olney JW, Zorumski CF, Stewart GR, Price MT, Wang G, and Labruyere J (1990) Excito-toxicity of L-DOPA and 6-OH-DOPA: Implications for Parkinson’s and Huntington’s diseases. Exp Neurol 108, 269–272
Rosenberg PA, Loring R, Xie Y, Zaleskas V, and Aizenman E (1991) 2,4,5-Trihydroxy-phenylalanine in solution forms a non N-methyl-D-aspartate glutamatergic agonist and neurotoxin. Proc Natl Acad Sci USA 88, 4865–4869
Skaper SD, Negro A, Facci L, and Dal Toso R (1993) Brain-derived neurotrophic factor selectively rescues mesencephalic dopaminergic neurons from 2,4,5-trihydroxyphenylalanine-induced injury. J Neurosci Res 34, 478–487
Acknowledgments
L. Facci was supported by “Fondazione CARIPARO Progetto Dottorati di Ricerca” Anno 2009.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2012 Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Skaper, S.D., Mercanti, G., Facci, L. (2012). Culture and Characterization of Rat Mesencephalic Dopaminergic Neurons. In: Skaper, S. (eds) Neurotrophic Factors. Methods in Molecular Biology, vol 846. Humana Press. https://doi.org/10.1007/978-1-61779-536-7_9
Download citation
DOI: https://doi.org/10.1007/978-1-61779-536-7_9
Published:
Publisher Name: Humana Press
Print ISBN: 978-1-61779-535-0
Online ISBN: 978-1-61779-536-7
eBook Packages: Springer Protocols