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Approaches to Recombinant Histamine H3/H4 Receptor Expression in Mammalian Cells

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Histamine Receptors as Drug Targets

Part of the book series: Methods in Pharmacology and Toxicology ((MIPT))

Abstract

Recombinant receptor techniques are very commonly utilized in pharmacological studies nowadays. This state of affairs is a result of recent development of genetic engineering and DNA cloning methods, which made the expression of foreign genes in various cellular environments possible. The starting point for the application of recombinant proteins in studies on seven-transmembrane receptors (7TMRs) was marked by cloning of the first two 7TMRs-cholinergic and adrenergic receptors in the late 1980s. Resolving of gene sequences for successive receptors enabled the use of expression techniques for already well-characterized 7TMRs, but also for orphan receptors. Within the toolbox of available systems for expression of recombinant receptors, mammalian cells constitute the most frequently used model, since they offer a high probability for maintenance of full functional activity of artificially overexpressed receptors. Taking into consideration the importance of mammalian expression systems in relation to current challenges of molecular pharmacology, some of the available protocols for recombinant 7TMRs production in mammalian cells will be discussed in this chapter.

In the first part, details of transient transfection of CHO cells with genetic construct encoding for fusion protein of histamine H3 receptor and fluorescent protein-mCherry using polyethylenimine as transfection reagent will be given. Then, methods for establishing a monoclonal cell lines stably expressing receptor protein following the lipofection procedure will be presented. Finally, the utility of retroviral expression systems for recombinant receptor production will be discussed using the histamine H4 receptor as an example.

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Acknowledgments

The authors acknowledge the financial support of Polish National Science Centre within the frame of Meastro grant No. 2011/02/A/NZ4/00031 and Preludium grant No. 2011/01/N/NZ4/01126, which enabled the development and optimization of protocols described in this manuscript.

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Authors and Affiliations

  1. Department of Technology and Biotechnology of Drugs, Faculty of Pharmacy, Jagiellonian University Medical College, Medyczna 9, 30-688, Kraków, Poland

    Tadeusz Karcz, Urszula Cichoń & Katarzyna Kieć-Kononowicz

Authors
  1. Tadeusz Karcz
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  2. Urszula Cichoń
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  3. Katarzyna Kieć-Kononowicz
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Corresponding author

Correspondence to Katarzyna Kieć-Kononowicz .

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Editors and Affiliations

  1. Department of Pharmacology Medical School, National and Kapodistrian University of Athens, Athens, Greece

    Ekaterini Tiligada

  2. Centre for Experimental Medicine, School of Medicine, Dentistry and Biomedical Sciences, Queen’s University Belfast, Belfast, Northern Ireland, United Kingdom

    Madeleine Ennis

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Karcz, T., Cichoń, U., Kieć-Kononowicz, K. (2017). Approaches to Recombinant Histamine H3/H4 Receptor Expression in Mammalian Cells. In: Tiligada, E., Ennis, M. (eds) Histamine Receptors as Drug Targets. Methods in Pharmacology and Toxicology. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6843-5_3

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  • DOI: https://doi.org/10.1007/978-1-4939-6843-5_3

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-6841-1

  • Online ISBN: 978-1-4939-6843-5

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