title: A pre-loading method of evaluating gap junctional communication by fluorescent dye transfer.
abstract: We describe a simple method for evaluating gap junctional communication (GJC) between cells in culture. The procedure involves pre-loading cells with two fluorescent dyes: calcein and DiI. Calcein is able to pass through gap junctions, while DiI is not. These pre-loaded cells are then plated with unlabeled cells. The number of cells receiving calcein from each pre-loaded cell can then be quantified after the cells settle on the plate. Potent and reversible inhibitors of GJC can be used in this system to evaluate dye transfer within a given period of time.
authors.affiliation: University of Western Ontario, London, Canada.
authors.forename: G S
authors.initials: GS
authors.lastname: Goldberg
authors.forename: J F
authors.initials: JF
authors.lastname: Bechberger
authors.forename: C C
authors.initials: CC
authors.lastname: Naus
mesh_terms: D002229:Carbenoxolone; D002232:Carbocyanines / Q000378:metabolism; D002450:Cell Communication / Q000187:drug effects / Q000502:physiology*; D002460:Cell Line; D002461:Cell Line, Transformed; D018031:Connexin 43 / Q000235:genetics; D005452:Fluoresceins / Q000378:metabolism; D005456:Fluorescent Dyes / Q000378:metabolism*; D017629:Gap Junctions / Q000378:metabolism*; D006034:Glycyrrhetinic Acid / Q000031:analogs & derivatives / Q000494:pharmacology; D008856:Microscopy, Fluorescence; D015688:Oncogene Protein pp60(v-src) / Q000235:genetics; D013237:Stereoisomerism; D013386:Succinates / Q000494:pharmacology
chemical_list: D002232:Carbocyanines; D018031:Connexin 43; D005452:Fluoresceins; D005456:Fluorescent Dyes; D013386:Succinates; C024286:3,3'-dioctadecylindocarbocyanine; D015688:Oncogene Protein pp60(v-src); D002229:Carbenoxolone; D006034:Glycyrrhetinic Acid; C007740:fluorexon