Heterologous Expression and Patch-Clamp Recording of P2X Receptors in HEK293 Cells

  • Lin-Hua JiangEmail author
  • Sébastien Roger
Part of the Methods in Molecular Biology book series (MIMB, volume 2041)


P2X receptors (P2XRs) are ligand-gated ion channels gated by extracellular adenosine 5′-triphosphate (ATP) and play a critical role in mediating ATP-induced purinergic signaling in physiological and pathological processes. Heterologous expression of P2XR in human embryonic kidney 293 (HEK293) cells and measurement of P2XR-mediated currents using patch-clamp recording technique have been widely used to study the biophysical and pharmacological properties of these receptors. Combination of electrophysiology with site-directed mutagenesis and structural information has shed light on the molecular basis for receptor activation and mechanisms of actions by receptor antagonists and modulators. It is anticipated that such methodologies will continue helping us to provide more mechanistic understanding of P2XRs and to test novel receptor antagonists and allosteric modulators for therapeutical purposes. In this chapter, we describe protocols of transiently or stably expressing the P2XR in HEK293 cells and measuring P2XR-mediated currents by using whole-cell recording.

Key words

P2X receptors HEK293 cells Heterologous expression Patch-clamp recording 



The work from Jiang’s laboratory was supported by Biotechnology and Biological Sciences Research Council and Wellcome Trust.


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© Springer Science+Business Media, LLC, part of Springer Nature 2020

Authors and Affiliations

  1. 1.School of Biomedical Sciences, Faculty of Biological SciencesUniversity of LeedsLeedsUK
  2. 2.EA4245, Transplantation, Immunology, Inflammation, Faculty of MedicineUniversity of ToursToursFrance

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