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Heterologous Expression of Ion Channels in Mammalian Cell Lines

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Patch Clamp Electrophysiology

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2188))

Abstract

Heterologous expression of recombinant ion channel subunits in mammalian cell lines allows for characterization of their functional properties and pharmacological regulation. In this chapter, we describe methods for thawing, refreezing, passaging, cell culture, and transfection of tsA201 cells suitable for electrophysiology and imaging experiments. Furthermore, we discuss the strengths and limitations of using these methods.

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References

  1. Chahine M, Plante E, Kallen RG (1996) Sea anemone toxin (ATX II) modulation of heart and skeletal muscle sodium channel alpha-subunits expressed in tsA201 cells. J Membr Biol 152:39–48

    Article  CAS  PubMed  Google Scholar 

  2. Shen ES, Cooke GM, Horlick RA (1995) Improved expression cloning using reporter genes and Epstein-Barr virus ori-containing vectors. Gene 156:235–239

    Article  CAS  PubMed  Google Scholar 

  3. Yang N, Ji S, Zhou M, Ptácek LJ, Barchi RL, Horn R, George AL Jr (1994) Sodium channel mutations in paramyotonia congenita exhibit similar biophysical phenotypes in vitro. Proc Natl Acad Sci U S A 91:12785–12789

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  4. Chahine M, Bennett PB, George AL Jr, Horn R (1994) Functional expression and properties of the human skeletal muscle sodium channel. Pflugers Arch 427:136–142

    Article  CAS  PubMed  Google Scholar 

  5. O’Leary ME, Horn R (1994) Internal block of human heart sodium channels by symmetrical tetra-alkylammoniums. J Gen Physiol 104:507–522

    Article  PubMed  Google Scholar 

  6. Margolskee RF, McHendry-Rinde B, Horn R (1993) Panning transfected cells for electrophysiological studies. Biotechniques 15:906–911

    CAS  PubMed  Google Scholar 

  7. Thomas P, Smart TG (2005) HEK293 cell line: a vehicle for the expression of recombinant proteins. J Pharmacol Toxicol Methods 51:187–200

    Article  CAS  PubMed  Google Scholar 

  8. Chen C, Okayama H (1987) High-efficiency transformation of mammalian cells by plasmid DNA. Mol Cell Biol 7:2745–2752

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  9. Veale EL, Al Moubarak E, Bajaria N, Omoto K, Cao L, Tucker SJ, Stevens EB, Mathie A (2014) Influence of the N-terminus on the biophysical properties and pharmacology of TREK1 potassium channels. Mol Pharmacol 85:671–681

    Article  PubMed  Google Scholar 

  10. Veale EL, Hassan M, Walsh Y, Al Moubarak E, Mathie A (2014) Recovery of current through mutated TASK3 potassium channels underlying Birk Barel syndrome. Mol Pharmacol 85:397–407

    Article  PubMed  Google Scholar 

  11. Veale EL, Mathie A (2016) Aristolochic acid, a plant extract used in the treatment of pain and linked to Balkan Endemic Nephropathy, is a regulator of K2P channels. Br J Pharmacol 173:1639–1652

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  12. Al-Moubarak E, Veale EL, Mathie A (2019) Pharmacologically reversible, loss of function mutations in the inner pore helices of TREK-1 K2P channels. Sci Rep 9:12394

    Google Scholar 

  13. Cunningham KP, Holden RG, Escribano-Subias P, Cogolludo A, Veale EL, Mathie A (2019) Characterisation and regulation of wild type and mutant TASK-1 two pore domain potassium channels indicated in pulmonary arterial hypertension. J Physiol 597:1087–1101

    Article  CAS  PubMed  Google Scholar 

  14. Cunningham KP, MacIntyre DE, Mathie A, Veale EL (2020) Effects of the ventilator stimulant, doxapram, on human TASK-3 (KCNK9, K2P9.1) and TASK-1 (KCNK3, K2P3.1) channels. Acta Physiol 228:e13361

    Google Scholar 

  15. Graham FL, van der Eb AJ (1973) A new technique for the assay of infectivity of human adenovirus 5 DNA. Virology 52:456–467

    Article  CAS  PubMed  Google Scholar 

  16. Graham FL, Smiley J, Russell WC, Nairn R (1977) Characteristics of a human cell line transformed by DNA from human adenovirus type 5. J Gen Virol 36:59–74

    Article  CAS  PubMed  Google Scholar 

  17. Batard P, Jordan M, Wurm F (2001) Transfer of high copy number plasmid into mammalian cells by calcium phosphate transfection. Gene 270:61–68

    Article  CAS  PubMed  Google Scholar 

  18. Chen CA, Okayama H (1988) Calcium phosphate-mediated gene transfer: a highly efficient transfection system for stably transforming cells with plasmid DNA. Biotechniques 6:632–638

    CAS  PubMed  Google Scholar 

  19. Veale EL, Rees KA, Mathie A, Trapp S (2010) Dominant negative effects of a non-functional TREK1 splice variant expressed in brain. J Biol Chem 285:29295–29304

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  20. El Hachmane MF, Rees KA, Veale EL, Sumbayev VV, Mathie A (2014) Enhancement of TWIK-related acid sensitive potassium channel 3 (TASK3) two pore domain potassium channel activity by TNFα. J Biol Chem 289:1388–1401

    Article  PubMed  Google Scholar 

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Authors and Affiliations

  1. Medway School of Pharmacy, University of Kent and University of Greenwich, Chatham Maritime, UK

    Alistair Mathie, Emma L. Veale & Robyn G. Holden

Authors
  1. Alistair Mathie
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  2. Emma L. Veale
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  3. Robyn G. Holden
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Corresponding author

Correspondence to Alistair Mathie .

Editor information

Editors and Affiliations

  1. University of Reading, Reading, UK

    Mark Dallas

  2. Sophion Bioscience A/S, Ballerup, Denmark

    Damian Bell

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Mathie, A., Veale, E.L., Holden, R.G. (2021). Heterologous Expression of Ion Channels in Mammalian Cell Lines. In: Dallas, M., Bell, D. (eds) Patch Clamp Electrophysiology. Methods in Molecular Biology, vol 2188. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0818-0_3

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  • DOI: https://doi.org/10.1007/978-1-0716-0818-0_3

  • Published:

  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-0817-3

  • Online ISBN: 978-1-0716-0818-0

  • eBook Packages: Springer Protocols

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