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.
Zidell
R H
RH
Department of Environmental and Industrial Health, University of Michigan School of Public Health, Ann Arbor 48109-2029.
Loch-Caruso
R
R
D002450:Cell Communication; D004396:Coloring Agents; D003513:Cycloheximide / Q000494:pharmacology; D003609:Dactinomycin / Q000494:pharmacology; D005260:Female; D006801:Humans; D007365:Intercellular Junctions / Q000502:physiology* / Q000648:ultrastructure; D007890:Leiomyosarcoma / Q000235:genetics / Q000473:pathology / Q000648:ultrastructure; D014176:Protein Biosynthesis / Q000187:drug effects; D011506:Proteins / Q000235:genetics; D014158:Transcription, Genetic / Q000187:drug effects; D014407:Tumor Cells, Cultured / Q000473:pathology / Q000648:ultrastructure; D014594:Uterine Neoplasms / Q000235:genetics / Q000473:pathology / Q000648:ultrastructure
D004396:Coloring Agents; D011506:Proteins; D003609:Dactinomycin; D003513:Cycloheximide