A simple dye-coupling assay for evaluating gap junctional communication: the importance of transcription and translation on the establishment of dye-coupling.

A simple dye-coupling assay has been developed to study gap junctional communication using a uterine cell line. The new dye-coupling assay was found to be of comparable sensitivity to an autoradiographic assay for transfer of radiolabelled nucleotide. To investigate the importance of protein synthesis in the establishment of dye-coupling under the conditions of this assay, cultures were treated with actinomycin-D and cycloheximide to inhibit transcription and translation, respectively. Inhibition of transcription had no significant effect on dye-coupling. The protein synthesis inhibitor, cycloheximide, also had no significant effect on dye-coupling at concentrations up to 1 uM, even though protein synthesis was inhibited to 19% of control.