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Using Whole-Cell Electrophysiology and Patch-Clamp Photometry to Characterize P2X7 Receptor Currents

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The P2X7 Receptor

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2510))

Abstract

The fundamental property of P2X7 receptor (P2X7R) channels is the transport of cations across the cell surface membrane. Electrophysiology and patch-clamp photometry are readily accessible methods of measuring this flux in a wide range of cell types. They are important tools used to characterize the functional properties of native cells studied in cell culture, in vitro tissue slices, and, in some cases, in situ single cells. Further, they are efficient methods of probing the relation of structure to function of recombinant receptors expressed in heterologous systems. Here, we provide step-by-step procedures for use of two standard recording protocols, broken-patch and perforated-patch voltage clamp. Further, we describe a third technique, called the dye-overload method, that uses simultaneous measurement of membrane current and fura-2 fluorescence to quantify the contribution of Ca2+ flux to the ATP-gated current.

Xin Liang, Laura Janks, and Terrance M. Egan are joint first authors.

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Abbreviations

AU:

Arbitrary unit

BU:

Bead unit

DMSO:

Dimethyl sulfoxide

ECS:

Extracellular solution

ICS:

Intracellular solution

P2XR:

P2X receptor

PCa/PNa:

Ca2+ Permeability relative to Na+

Pf%:

Fractional Ca2+ current

PMT:

Photomultiplier tube

ROI:

Region of interest

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Acknowledgments

Supported by grants from the NIH (TME, R01GM112188), Saint Louis University (TME), the Natural Science Foundation of Guangdong Province (XL, No. 2020A1515010802), Basic Research Program of Shenzhen (XL, JCYJ20180507181654186), Health system scientific research project of Shenzhen Guangming District Science and innovation Bureau (XL, 2020R01073), Special fund for economic development of ShenZhen Guangming District (XL, 2021R01128), Disciplinary Construction of Posts for Zhujiang Scholars (XL, 4SG21005G), and Discipline construction project of Guangdong Medical University (XL, 4SG21008G).

Author information

Author notes

  1. Laura Janks

    Present address: Checkpoint Immunology, Immunology and Inflammation Research Therapeutic Area, Sanofi, Cambridge, MA, USA

Authors and Affiliations

  1. The China-America Cancer Research Institute, School of Basic Medical Sciences, Guangdong Medical University, Dongguan, P. R. China

    Xin Liang

  2. Department of Pharmacology and Physiology, Saint Louis University School of Medicine, St. Louis, MO, USA

    Laura Janks & Terrance M. Egan

  3. Henry and Amelia Nasrallah Center for Neuroscience, Saint Louis University School of Medicine, St. Louis, MO, USA

    Laura Janks & Terrance M. Egan

Authors
  1. Xin Liang
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  2. Laura Janks
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  3. Terrance M. Egan
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Corresponding author

Correspondence to Terrance M. Egan .

Editor information

Editors and Affiliations

  1. Walther Straub Institute of Pharmacology and Toxicology, Ludwig-Maximilians-Universität München, Munich, Germany

    Annette Nicke

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Liang, X., Janks, L., Egan, T.M. (2022). Using Whole-Cell Electrophysiology and Patch-Clamp Photometry to Characterize P2X7 Receptor Currents. In: Nicke, A. (eds) The P2X7 Receptor. Methods in Molecular Biology, vol 2510. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2384-8_11

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  • DOI: https://doi.org/10.1007/978-1-0716-2384-8_11

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-2383-1

  • Online ISBN: 978-1-0716-2384-8

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