Abstract
Ion channels are membrane proteins that gate the flow of ions into and out of a cell. They are present in the membranes of human, animal, plant, and bacterial cells. They are profoundly involved in diverse tasks ranging from neuronal functions to hormonal secretion and cell division. Biophysical characterization and modulation of ion channel targets are important approaches in modern drug discovery. With the heterologous expression of the nicotinic acetylcholine receptor alpha7 (nAChRα7) in a host cell, we show a way to construct and use such a stable cell-based expression system for electrophysiological assays.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Ashcroft F (2000) Ion Channels and Disease. Academic Press, San Diego, CA, p 481
Venter JC, Adams MD, Myers EW, Li PW et al (2001) The sequence of the human genome. Science 291:1304–1351
Li S, Gosling M, Poll CT, Westwick J, Cox B (2005) Therapeutic scope of modulation of non-voltage-gated cation channels. Drug Discov Today 10:129–137
Hamill OP, Marty A, Neher E, Sakmann B, Sigworth FJ (1981) Improved patch-clamp techniques for high-resolution current recording from cells and cell-free membrane patches. Pflugers Arch 391:85–100
Fermini B, Fossa AA (2003) The impact of drug-induced QT interval prolongation on drug discovery and development. Nat Rev Drug Discov 2:439–447
Xu J, Wang X, Ensign B, Li M, Wu L, Guia A, Xu J (2001) Ion-channel assay technologies: quo vadis? Drug Discov Today 6:1278–1287
Wang X, Li M (2003) Automated electrophysiology: high throughput of art. Assay Drug Dev Technol 1:695–708
Dunlop J, Bowlby M, Peri R, Vasilyev D, Arias R (2008) High-throughput electrophysiology: an emerging paradigm for ion-channel screening and physiology. Nat Rev Drug Discov 7:358–368
Bianchi BR, Moreland RB, Faltynek CR, Chen J (2007) Application of large-scale transiently transfected cells to functional assays of ion channels: different targets and assay formats. Assay Drug Dev Technol 5:417–424
Deutsch C (2003) The birth of a channel. Neuron 40:265–276
Jenkinson DH (2006) Potassium channels-multiplicity and challenges. Br J Pharmacol 147(Suppl 1):63–71
Feuerbach D, Lingenhoehl K, Olpe HR, Vassout A, Gentsch C, Chaperon F, Nozulak J, Enz A, Bilbe G, McAllister K, Hoyer D (2009) The selective nicotinic acetylcholine receptor alpha7 agonist JN403 is active in animal models of cognition, sensory gating, epilepsy and pain. Neuropharmacology 56:254–256
Lyford LK, Rosenberg RL (1999) Cell-free expression and functional reconstitution of homo-oligomeric alpha7 nicotinic acetylcholine receptors into planar lipid bilayers. J Biol Chem 274:25675–25681
Treinin M (2008) RIC-3 and nicotinic acetylcholine receptors: biogenesis, properties, and diversity. Biotechnol J 3:1539–1547
Zwart R, Vijverberg HP (1997) Potentiation and inhibition of neuronal nicotinic receptors by atropine: competitive and noncompetitive effects. Mol Pharmacol 52:886–895
Gault J, Robinson M, Berger R, Drebing C et al (1998) Genomic organization and partial duplication of the human alpha7 neuronal nicotinic acetylcholine receptor gene (CHRNA7). Genomics 52:173–185
Verdoorn TA, Draguhn A, Ymer S, Seeburg PH, Sakmann B (1990) Functional properties of recombinant rat GABAA receptors depend upon subunit composition. Neuron 4:919–928
Acknowledgments
The authors thank Céline Wimmersberger for technical assistance.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2010 Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Steiner, F., Ghose, S., Thomet, U. (2010). Recombinant Cell Lines Stably Expressing Functional Ion Channels. In: Szallasi, A. (eds) Analgesia. Methods in Molecular Biology, vol 617. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-60327-323-7_17
Download citation
DOI: https://doi.org/10.1007/978-1-60327-323-7_17
Published:
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-60327-322-0
Online ISBN: 978-1-60327-323-7
eBook Packages: Springer Protocols