Abstract
The identification of specific and selective markers of the dopamine-producing neurons that are lost in Parkinson’s disease has been a major research focus since Hornykiewicz first reported a dopamine deficiency in the disease (1). Antibodies to dopamine or tyrosine hydroxylase, the rate-limiting enzyme in dopamine synthesis, have been used to identify these neurons. Recently, considerable attention has been given to the plasma membrane dopamine tranporter (DAT) and the vesicular monoamine transporter (VMAT2), which are responsible for the transport, packaging, and release of dopamine (2). DAT acts to terminate dopamine transmission by rapid reuptake of dopamine from the synapse, and VMAT2 packages cytoplasmic dopamine into vesicles for storage and subsequent release. We have developed specific antibodies to these transporters and used them to characterize the distribution and expression of DAT and VMAT2 in brain from human idiopathic Parkinson’s disease and animal models of the disease. The purpose of this chapter is to describe the immunochemical techniques involved in assessing damage to dopamine neurons in Parkinson’s disease and experimental models of the disease.
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References
Ehringer, H. H. (1960) Distribution of noradrenaline and dopamine (3-hydroxytyramine)in the human brain and their behavior in diseases of the extrapyramidal system. Klin. Wochenschr 38, 1236–1239.
Miller, G. W., Gainetdinov, R. R., Levey, A. I., and Caron, M. G. (1999) Dopamine transporters and neuronal injury. Trends Pharmacol. Sci. 20B 424–429.
Wilson, J. M., Levey, A. I., Rajput, A., Ang, L., Guttman, M., Shannak, K., et al. (1996) Differential changes in neurochemical markers of striatal dopamine nerve terminals in idiopathic Parkinson’s disease. Neurology 47, 718–726.
Miller, G. W., Staley, J. K., Heilman, C. J., Perez, J. T., Mash, D. C., Rye, D. B., et al.(1997) Immunochemical analysis of dopamine transporter protein in Parkinson’s disease. Ann. Neurol. 41, 530–539.
Miller, G. W., Erickson, J. D., Perez, J. T., Penland, S. N., Mash, D. C., Rye, D. B., et al. (1999) Immunochemical analysis of vesicular monoamine transporter (VMAT2) protein in Parkinson’s disease. Exp. Neurol. 154, 138–148.
Miller, G. W., Gilmor, M. L., and Levey, A.I. (1998) Generation of transporter specific antibodies. Meth. Enzymol. 296, 407–422.
Polak, J. M. (1977) Introduction to Immunocytochemistry. Springer, Oxford.
Javois, L. C., ed. (1999) Immunocytochemistry Methods and Protocols, 2nd ed. Humana, Totowa, NJ.
Beesley, J. E., ed. (1993) Immunocytochemistry: A Practical Approach. Oxford University Press, Oxford.
Ciliax, B. J., Drash, G. W., Staley, J. K., Haber, S., Mobley, C. J., Miller, G. W., et al. (1999) Immunocytochemical localization of the dopamine transporter in human brain. J. Comp. Neurol. 409, 38–56.
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© 2001 Humana Press Inc.
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Miller, G.W., Levey, A.I. (2001). Immunochemical Analysis of Dopamine Transporters in Parkinson's Disease. In: Maral Mouradian, M. (eds) Parkinson's Disease. Methods in Molecular Medicine™, vol 62. Humana Press. https://doi.org/10.1385/1-59259-142-6:167
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DOI: https://doi.org/10.1385/1-59259-142-6:167
Publisher Name: Humana Press
Print ISBN: 978-0-89603-761-8
Online ISBN: 978-1-59259-142-8
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