A pre-loading method of evaluating gap junctional communication by fluorescent dye transfer.
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.
Goldberg
G S
GS
University of Western Ontario, London, Canada.
Bechberger
J F
JF
Naus
C C
CC
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
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