Abstract
TaqMan reverse transcription-polymerase chain reaction (TaqMan RT-PCR) is a recently developed technique (1) that has been used to study gene expression in tissues of the central nervous system (CNS) including the striatum. For example, TaqMan has been used to profile mRNA distribution patterns across the brain for γ-aminobutyric acid-B (GABAB) receptor subunits (2), 5-hydroxytryptamine4 (5-HT4) receptor splice variants (3), novel G-protein-coupled receptors (4), and ion channels including vanilloid receptors (5) and two pore potassium channels (6). More specifically, TaqMan RT-PCR studies have demonstrated a huge enrichment of dopamine D2 (7) and D3 (8) receptors in human striatal tissues compared to other brain regions. In addition, the technique has been utilized for the analysis of gene expression changes in animal models of CNS diseases including Parkinsons’s disease (9), stroke (10), and migraine (7).
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References
Heid, C. A., Stevens, J., Livak, K. J., and Williams, P. M. (1996) Real time quantitative PCR. Genome Methods 6, 986–994.
Calver, A. R., Medhurst, A. D., Robbins, M. J., et al. (2000) Expression of GABAB1 and GABAB2 receptor subunits in the central nervous system differs from that in peripheral tissues. Neuroscience 100, 155–170.
Medhurst, A. D., Lezoualch, F., Fischmeister, R., Middlemiss, D. N., and Sanger, G. J. (2001) Quantitative mRNA analysis of five c-terminal splice variants of the human 5-HT4 receptor in the central nervous system by TaqMan real time RT-PCR. Mol. Brain Res. 90, 125–134.
Robbins, M. J., Michalovich, D., Hill, J., et al. (2000) Molecular cloning and characterisation of two novel retinoic acid inducible orphan G protein coupled receptors. Genomics 67, 8–18.
Hayes, P., Meadows, H., Harries, M., et al. (2000) Cloning and expression of a human homologue to rat vanilloid receptor-1. Pain 88, 205–215.
Medhurst, A. D., Rennie, G., Chapman, C. G., et al. (2001) Distribution analysis of human two pore domain potassium channels in tissues of the central nervous system and periphery. Mol. Brain Res. 86, 101–114.
Medhurst, A. D., Harrison, D. C., Read, S. J, Campbell, C. A., Robbins, M. J., and Pangalos, M. N. (2000) The use of TaqMan RT-PCR assays for semiquantitative analysis of gene expression in CNS tissues and disease models. J. Neurosci. Methods 98, 9–20.
Reavill, C., Taylor, S. G., Wood, M. D., et al. (2000) Pharmacological actions of a novel, high-affinity, and selective human dopamine D(3) receptor antagonist, SB-277011-A. J. Pharmacol. Exp. Ther. 294, 1154–1165.
Medhurst, A. D., Zeng, B. Y., Charles, K. J., et al. (2001) Up-regulation of secretoneurin immunoreactivity and secretogranin II mRNA in rat striatum following 6-hydroxydopamine lesioning and chronic L-DOPA treatment. Neuroscience 105, 353–364.
Harrison, D. C., Medhurst, A. D., Bond, B. C., Campbell, C. A., Davis, R. P., and Philpott, K. L. (2000) The use of quantitative RT-PCR to measure mRNA expression in a rat model of focal ischemia—caspase-3 as a case study. Mol. Brain Res. 75, 143–149.
Lang, R., Pfeffer, K., Wagner, H., and Heeg, K. (1997) A rapid method for semiquantitative analysis of the human Vβ-repertoire using taqMan PCR. J. Immunol. Methods 203, 181–192.
Lie, Y. S. and Petropoulos, C. J. (1998) Advances in quantitative PCR technology: 5′ nuclease assays. Curr. Opin. Biotech. 9, 43–48.
Livak, K. J. (1999) Allelic discrimination using fluorogenic probes and the 5′-nuclease assay. Gen. Analyt. Biomol. Engin. 14, 143–149.
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© 2003 Humana Press Inc., Totowa, NJ
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Medhurst, A.D., Pangalos, M.N. (2003). Application of TaqMan RT-PCR for Real-Time Semiquantitative Analysis of Gene Expression in the Striatum. In: Wang, J.Q. (eds) Drugs of Abuse. Methods In Molecular Medicine™, vol 79. Humana Press. https://doi.org/10.1385/1-59259-358-5:229
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DOI: https://doi.org/10.1385/1-59259-358-5:229
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Online ISBN: 978-1-59259-358-3
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