title: Electroporation Loading and Dye Transfer: A Safe and Robust Method to Probe Gap Junctional Coupling.
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.
authors.affiliation: Physiology Group, Department of Basic Medical Sciences, Ghent University, De Pintelaan 185, Ghent, 9000, Belgium.
authors.forename: Elke
authors.initials: E
authors.lastname: Decrock
authors.affiliation: Physiology Group, Department of Basic Medical Sciences, Ghent University, De Pintelaan 185, Ghent, 9000, Belgium.
authors.forename: Marijke
authors.initials: M
authors.lastname: De Bock
authors.affiliation: Physiology Group, Department of Basic Medical Sciences, Ghent University, De Pintelaan 185, Ghent, 9000, Belgium.
authors.forename: Diego
authors.initials: D
authors.lastname: De Baere
authors.affiliation: Physiology Group, Department of Basic Medical Sciences, Ghent University, De Pintelaan 185, Ghent, 9000, Belgium.
authors.forename: Delphine
authors.initials: D
authors.lastname: Hoorelbeke
authors.affiliation: Physiology Group, Department of Basic Medical Sciences, Ghent University, De Pintelaan 185, Ghent, 9000, Belgium.
authors.forename: Nan
authors.initials: N
authors.lastname: Wang
authors.affiliation: Physiology Group, Department of Basic Medical Sciences, Ghent University, De Pintelaan 185, Ghent, 9000, Belgium. luc.leybaert@ugent.be.
authors.forename: Luc
authors.initials: L
authors.lastname: Leybaert
mesh_terms: D000818:Animals; D002229:Carbenoxolone / Q000494:pharmacology; D002450:Cell Communication / Q000502:physiology; D002452:Cell Count; D045744:Cell Line, Tumor; D002470:Cell Survival; D018031:Connexin 43 / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D018274:Electroporation / Q000295:instrumentation / Q000379:methods*; D005456:Fluorescent Dyes / Q000009:adverse effects / Q000378:metabolism*; D017629:Gap Junctions / Q000502:physiology*; D006801:Humans; D008856:Microscopy, Fluorescence; D008858:Microscopy, Phase-Contrast / Q000379:methods; D051381:Rats; D013194:Staining and Labeling / Q000379:methods*; D014162:Transfection / Q000379:methods*
chemical_list: D018031:Connexin 43; D005456:Fluorescent Dyes; D002229:Carbenoxolone
keywords: Connexin; Dye transfer; Electroporation; Gap junction; Intercellular communication; Time-lapse imaging