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Receptor Signal Transduction Protocols pp 165–193Cite as

Quantification of GPCR mRNA Using Real-Time RT-PCR

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 746))

Abstract

Characterisation of G-protein-coupled receptor (GPCR) mRNA expression under normal, different pharmacological and pathological conditions in experimental animal models and human tissue biopsies by quantitative real-time reverse transcription polymerase chain reaction (RT-qPCR) is a valuable approach to understand the regulation of GPCR expression. RT-qPCR is specific and sensitive with a broad dynamic range, which allows precise quantification of mRNA species of interest. In addition to measuring the relative levels of mRNA in a tissue or changes in expression levels between groups of genes of interest, RT-qPCR is also used to identify splice variants and single nucleotide polymorphisms (SNPs) of GPCRs. Even though RT-qPCR has become the standard method for quantification of gene expression, RT-qPCR is sensitive to RNA quality, assay design, normalisation approach and data analysis. This protocol is meant as a guide to RT-qPCR methodology with references to the best standard methods available at present.

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Acknowledgements

Work in the authors’ laboratories has been supported by The Norwegian Council on Cardiovascular Disease, The Research Council of Norway, Anders Jahre’s Foundation for the promotion of Science, The Novo Nordisk Foundation, The Family Blix Foundation and Shipowner Tom Wilhelmsen’s Foundation. Anne Vatland Krøvel is acknowledged for critical reading of the manuscript.

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Authors and Affiliations

  1. Department of Pharmacology, Center for Heart Failure Research, University of Oslo and Oslo University Hospital, Oslo, Norway

    Finn Olav Levy

Authors
  1. Trond Brattelid
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  2. Finn Olav Levy
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Corresponding author

Correspondence to Finn Olav Levy .

Editor information

Editors and Affiliations

  1. , Dept. Cell Physiology & Pharmacology, University Leicester, University Road, Leicester, LE1 9HN, United Kingdom

    Gary B. Willars

  2. , Dept. Cell Physiology & Pharmacology, University Leicester, University Road, Leicester, LE1 9HN, United Kingdom

    R.A. John Challiss

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Brattelid, T., Levy, F.O. (2011). Quantification of GPCR mRNA Using Real-Time RT-PCR. In: Willars, G., Challiss, R. (eds) Receptor Signal Transduction Protocols. Methods in Molecular Biology, vol 746. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-61779-126-0_9

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  • DOI: https://doi.org/10.1007/978-1-61779-126-0_9

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-61779-125-3

  • Online ISBN: 978-1-61779-126-0

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