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TRP Channels pp 99-110 | Cite as

Fluorescence-Based Functional Assays for Ca2+-Permeable ThermoTRP Channels

  • Celia Cordero-Sánchez
  • Irene Mudarra-Fraguas
  • Asia Fernández-CarvajalEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1987)

Abstract

Transient receptor potential (TRP) ion channels are involved in a variety of fundamental physiological processes, and their malfunction produces numerous human diseases. Therefore, these proteins represent a class of attractive drug targets and a class of important off-targets for in vitro pharmacological profiling. In the past decades, the rapid progress in emerging functional assays and instrumentation has enabled to readily monitor thermoTRP channel activity, and to develop high throughput screening (HTS) assays for TPR drug discovery. Chronologically, functional methods for ion channels include the ligand binding assay, flux-based assay, electrophysiology, fluorescence-based assays, and, more recently, automated electrophysiological assays. Here we described the methodology used to monitor the functionality of two thermoTRPs, TRPV1 and TRPM8, based on Ca2+ microfluorography using a 96-well fluorescence plate reader that allows the implementation of a medium- to high-throughput format ideal for drug screening.

Key words

Ion channels High-throughput screening Ligand binding assay Flux-based assay Fluorescence-based assay Drug discovery 

Notes

Acknowledgments

This work was supported by the Ministry of Economy and Competitiveness (SAF2016-66275-C02-01) and Generalitat Valenciana PROMETEO/2014/011.

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Celia Cordero-Sánchez
    • 1
  • Irene Mudarra-Fraguas
    • 1
  • Asia Fernández-Carvajal
    • 1
    Email author
  1. 1.Instituto de Investigación, Desarrollo e Innovación en Biotecnología Sanitaria de Elche (IDiBE)Universitas Miguel HernándezElcheSpain

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