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Simultaneous Real-Time Measurement of the β-Cell Membrane Potential and Ca2+ Influx to Assess the Role of Potassium Channels on β-Cell Function

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Potassium Channels

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1684))

Abstract

Stimulus-secretion coupling in pancreatic β-cells requires Ca2+ influx through voltage-dependent Ca2+ channels, whose activity is controlled by the plasma membrane potential (V m). Here, we present a method of measuring fluctuations in the β-cell V m and Ca2+ influx simultaneously, which provides valuable information about the ionic signaling mechanisms that underlie insulin secretion. This chapter describes the use of perforated patch clamp electrophysiology on cells loaded with a fluorescent intracellular Ca2+ indicator, which permits the stable recording conditions needed to monitor the V m and Ca2+ influx in β-cells. Moreover, this chapter describes the protocols necessary for the preparation of mouse and human islet cells for the simultaneous recording of V m and Ca2+ as well as determining the specific islet cell type assessed in each experiment.

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References

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Acknowledgments

Work in the laboratory of D.A.J. has been supported by National Institutes of Health Grants K01DK081666, R01DK097392, Vanderbilt DRTC Pilot and Feasibility Grant P60DK20593, ADA grant 1-17-IBS-024 (D.A.J.).; Vanderbilt METP grant 5T32DK07563, National Institutes of Health Grant 1F31DK109625 (N.C.V.); Vanderbilt ITED grant T32DK101003 (M.T.D.). Previous work in the Philipson lab was also supported by R01DK092616-01A1 (P.I.M.W. Roe) and P30DK020595 (G.I. Bell) and R01DK48494 (L.H. Philipson).

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Authors and Affiliations

  1. Department of Molecular Physiology and Biophysics, Vanderbilt University, Nashville, TN, 37232, USA

    Nicholas C. Vierra, Matthew T. Dickerson & David A. Jacobson

  2. Department of Medicine, Kovler Diabetes Center, The University of Chicago, Chicago, IL, USA

    Louis H. Philipson

  3. Department of Pediatrics, The University of Chicago, Chicago, IL, USA

    Louis H. Philipson

Authors
  1. Nicholas C. Vierra
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  2. Matthew T. Dickerson
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  3. Louis H. Philipson
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  4. David A. Jacobson
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Corresponding author

Correspondence to David A. Jacobson .

Editor information

Editors and Affiliations

  1. Department of Physiology and Pharmacology, Oregon Health and Science University, Portland, Oregon, USA

    Show-Ling Shyng

  2. Department of Physiology and Pharmacology, Oregon Health and Science University, Portland, Oregon, USA

    Francis I. Valiyaveetil

  3. The Vollum Institute, Oregon Health and Science University, Portland, Oregon, USA

    Matt Whorton

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Vierra, N.C., Dickerson, M.T., Philipson, L.H., Jacobson, D.A. (2018). Simultaneous Real-Time Measurement of the β-Cell Membrane Potential and Ca2+ Influx to Assess the Role of Potassium Channels on β-Cell Function. In: Shyng, SL., Valiyaveetil, F., Whorton, M. (eds) Potassium Channels. Methods in Molecular Biology, vol 1684. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7362-0_7

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  • DOI: https://doi.org/10.1007/978-1-4939-7362-0_7

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7361-3

  • Online ISBN: 978-1-4939-7362-0

  • eBook Packages: Springer Protocols

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