Gap junction-mediated intercellular communication in primary cultures of rainbow trout hepatocytes

Gap junction-mediated intercellular communication was evaluated in primary cultures of rainbow trout hepatocytes by measurement of dye coupling. Donor hepatocytes were microinjected with fluorescent Lucifer yellow CH dye and visualization of dye spread (dye coupling) to adjacent hepatocytes was recorded. A maximum level was reached between 8 and 12 hr which was maintained up to 24 hr. Dye coupling then decreased over the next 48 hr. Cell viability was monitored by the percentage of total LDH released and trypan blue exclusion at each time point. The effect of 12-O-tetradecanoylphorbol-13-acetate (TPA), a known inhibitor of rodent hepatic intercellular communication, on gap junction-mediated intercellular communication was evaluated in 24-hr cultures. A dose-response inhibition was demonstrated with maximum inhibition observed at 3 hr of exposure.