A pre-loading method of evaluating gap junctional communication by fluorescent dye transfer

Biotechniques. 1995 Mar;18(3):490-7.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Carbenoxolone
  • Carbocyanines / metabolism
  • Cell Communication / drug effects
  • Cell Communication / physiology*
  • Cell Line
  • Cell Line, Transformed
  • Connexin 43 / genetics
  • Fluoresceins / metabolism
  • Fluorescent Dyes / metabolism*
  • Gap Junctions / metabolism*
  • Glycyrrhetinic Acid / analogs & derivatives
  • Glycyrrhetinic Acid / pharmacology
  • Microscopy, Fluorescence
  • Oncogene Protein pp60(v-src) / genetics
  • Stereoisomerism
  • Succinates / pharmacology

Substances

  • Carbocyanines
  • Connexin 43
  • Fluoresceins
  • Fluorescent Dyes
  • Succinates
  • 3,3'-dioctadecylindocarbocyanine
  • Oncogene Protein pp60(v-src)
  • Carbenoxolone
  • Glycyrrhetinic Acid
  • fluorexon