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Electroporation Loading and Dye Transfer: A Safe and Robust Method to Probe Gap Junctional Coupling

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Gap Junction Protocols

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1437))

Abstract

Intercellular communication occurring via gap junction channels is considered a key mechanism for synchronizing physiological functions of cells and for the maintenance of tissue homeostasis. Gap junction channels are protein channels that are situated between neighboring cells and that provide a direct, yet selective route for the passage of small hydrophilic biomolecules and ions. Here, an electroporation method is described to load a localized area within an adherent cell monolayer with a gap junction-permeable fluorescent reporter dye. The technique results in a rapid and efficient labeling of a small patch of cells within the cell culture, without affecting cellular viability. Dynamic and quantitative information on gap junctional communication can subsequently be extracted by tracing the intercellular movement of the dye via time-lapse microscopy.

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Acknowledgements

This work is supported by the Fund for Scientific Research Flanders (FWO-Vlaanderen), Belgium (grants G.0298.11, G.0571.12, G.0A54.13, and G.0320.15), the Special Research Fund (BOF) of Ghent University (grant 01IO8314), and the Interuniversity Attraction Poles Program (Belgian Science Policy, project P7/10).

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Authors and Affiliations

  1. Physiology Group, Department of Basic Medical Sciences, Ghent University, De Pintelaan 185, Ghent, 9000, Belgium

    Elke Decrock, Marijke De Bock, Diego De Baere, Delphine Hoorelbeke, Nan Wang & Luc Leybaert

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  1. Elke Decrock
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  2. Marijke De Bock
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  3. Diego De Baere
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  4. Delphine Hoorelbeke
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  5. Nan Wang
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  6. Luc Leybaert
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Corresponding author

Correspondence to Luc Leybaert .

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Editors and Affiliations

  1. Department of in Vitro Toxicology and D, Vrije Universiteit Brussel, Brussels, Belgium

    Mathieu Vinken

  2. BHF Glasgow Cardiovascular Research, University of Glasgow, Glasgow, United Kingdom

    Scott R. Johnstone

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Decrock, E., De Bock, M., De Baere, D., Hoorelbeke, D., Wang, N., Leybaert, L. (2016). Electroporation Loading and Dye Transfer: A Safe and Robust Method to Probe Gap Junctional Coupling. In: Vinken, M., Johnstone, S. (eds) Gap Junction Protocols. Methods in Molecular Biology, vol 1437. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3664-9_11

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  • DOI: https://doi.org/10.1007/978-1-4939-3664-9_11

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-3662-5

  • Online ISBN: 978-1-4939-3664-9

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