Chapter title |
Electroporation Loading and Dye Transfer: A Safe and Robust Method to Probe Gap Junctional Coupling.
|
---|---|
Chapter number | 11 |
Book title |
Gap Junction Protocols
|
Published in |
Methods in molecular biology, January 2016
|
DOI | 10.1007/978-1-4939-3664-9_11 |
Pubmed ID | |
Book ISBNs |
978-1-4939-3662-5, 978-1-4939-3664-9
|
Authors |
Elke Decrock, Marijke De Bock, Diego De Baere, Delphine Hoorelbeke, Nan Wang, Luc Leybaert |
Editors |
Mathieu Vinken, Scott R. Johnstone |
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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