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Biophysical Methods to Analyze Direct G-Protein Regulation of Neuronal Voltage-Gated Calcium Channels

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Receptor and Ion Channel Detection in the Brain

Part of the book series: Neuromethods ((NM,volume 110))

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Abstract

Neuronal voltage-gated calcium channels play an essential role for calcium entry into presynaptic endings responsible for the release of neurotransmitters. In turn, and in order to fine tune synaptic activity, numerous neurotransmitters exert a potent negative feedback over the calcium signal provided by G-protein-coupled receptors that can be recognized by characteristic biophysical modifications of the calcium current. There are two main biophysical approaches to analyze direct G-protein regulation of voltage-gated calcium channels: the so-called double-pulse method, which is indirectly assessed by the gain of current produced by a depolarizing prepulse potential, and the “subtraction” method that allows the analysis of G-protein regulation from the ionic currents induced by regular depolarizing pulses. The later method separates the ionic currents due to nonregulated channels from the ion currents that result from a progressive departure of G-proteins from regulated channels, thereby providing valuable information on the OFF kinetics of G-protein regulation. In this chapter, we introduce these “double pulses” and “subtraction” procedures for use primarily with single cells and also discuss the limitations inherent to these two approaches.

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Acknowledgments

Research in NW’s laboratory is supported by the Czech Science Foundation (grant 15-13556S), the Ministry of Education Youth and Sports (grant 7AMB15FR015), and the Institute of Organic Chemistry and Biochemistry.

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Authors and Affiliations

  1. Institute of Organic Chemistry and Biochemistry, Academy of Sciences of the Czech Republic, v.v.i., Prague, Czech Republic

    Norbert Weiss

  2. Grenoble Institute of Neuroscience, University Joseph Fourier, Grenoble, France

    Michel De Waard

Authors
  1. Norbert Weiss
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  2. Michel De Waard
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Corresponding authors

Correspondence to Norbert Weiss or Michel De Waard .

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Editors and Affiliations

  1. Research Institute on Neurological Disabilities (IDINE), Department of Medical Sciences, School of Medicine, University of Castilla- La Mancha, Albacete, Spain

    Rafael Luján

  2. Pharmacology Unit, Department of Pathology and Experimental Therapeutics, School of Medicine, IDIBELL, University of Barcelona, Barcelona, Spain

    Francisco Ciruela

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Weiss, N., De Waard, M. (2016). Biophysical Methods to Analyze Direct G-Protein Regulation of Neuronal Voltage-Gated Calcium Channels. In: Luján, R., Ciruela, F. (eds) Receptor and Ion Channel Detection in the Brain. Neuromethods, vol 110. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3064-7_22

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  • DOI: https://doi.org/10.1007/978-1-4939-3064-7_22

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-3063-0

  • Online ISBN: 978-1-4939-3064-7

  • eBook Packages: Springer Protocols

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