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Methods for Investigating TRP Channel Gating

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TRP Channels

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1987))

Abstract

A complete characterization of temperature -and voltage-activated TRP channel gating requires a precise determination of the absolute probability of opening in a wide range of voltages, temperatures, and agonist concentrations. We have achieved this in the case of the TRPM8 channel expressed in Xenopus laevis oocytes. Measurements covered an extensive range of probabilities and unprecedented applied voltages up to 500 mV. In this chapter, we describe animal care protocols of patch-clamp pipette preparation, temperature control methods, and analysis of ionic currents to obtain reliable absolute open channel probabilities.

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References

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Acknowledgments

We thank Miss Luisa Soto for excellent technical assistance, and Dr. Juan Pablo Castillo for his help to the noise analysis method. This work was supported by FONDECYT Grants 1150273 and 1190293 (to R.L.), 1180464 (to C.G.), 1180999 (to K.C), CONICYT-PFCHA/Doctorado Nacional/2017-21170395 (to E.C.). The Centro Interdisciplinario de Neurociencia de Valparaíso is a Millennium Institute (P09-022-F). This work was partially supported by the Air Force Office of Scientific Research under award number FA9550-16-1-0384 to R.L.

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Authors and Affiliations

  1. Departamento de Biología, Facultad de Ciencias, Universidad de Chile, Santiago, Chile

    Osvaldo Alvarez

  2. Facultad de Ciencias, Centro Interdisciplinario de Neurociencia de Valparaíso, Universidad de Valparaíso, Valparaíso, Chile

    Osvaldo Alvarez, Karen Castillo, Emerson Carmona, Carlos Gonzalez & Ramon Latorre

Authors
  1. Osvaldo Alvarez
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  2. Karen Castillo
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  3. Emerson Carmona
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  4. Carlos Gonzalez
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  5. Ramon Latorre
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Corresponding author

Correspondence to Osvaldo Alvarez .

Editor information

Editors and Affiliations

  1. Instituto de Biología, Molecular y Celular, Universidad Miguel Hernández, Elche, Alicante, Spain

    Antonio Ferrer-Montiel

  2. Experimental Anesthesiology and Pain Research, Department of Anesthesiology and Intensive Care Medicine, University Hospital Cologne, University of Cologne, Cologne, Germany

    Tim Hucho

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Alvarez, O., Castillo, K., Carmona, E., Gonzalez, C., Latorre, R. (2019). Methods for Investigating TRP Channel Gating. In: Ferrer-Montiel, A., Hucho, T. (eds) TRP Channels. Methods in Molecular Biology, vol 1987. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9446-5_11

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  • DOI: https://doi.org/10.1007/978-1-4939-9446-5_11

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-4939-9445-8

  • Online ISBN: 978-1-4939-9446-5

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