GAP JUNCTION-MEDIATED INTERCELLULAR COMMUNICATION IN PRIMARY CULTURESOF RAINBOW-TROUT HEPATOCYTES

Gap junction-mediated intercellular communication was evaluated in Pri mary cultures of rainbow trout hepatocytes by measurement of dye coupl ing. Donor hepatocytes were microinjected with fluorescent Lucifer yel low 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 tot al LDH released and trypan blue exclusion at each time point. The effe ct 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-re sponse inhibition was demonstrated with maximum inhibition observed at 3 hr of exposure. (C) 1994 Academic Press, Inc.