Potassium transfer assay for cell communication: effects of phorbol esters, retinoic acid, and furosemide.
Using a mass culture assay for the contact-dependent transfer of potassium among cells with intrinsic differences in ability to concentrate it, we have investigated the ability of several drugs to influence this form of cell communication. We concentrated on 12-O-tetradecanoylphorbol-13-acetate (TPA), which is known to interfere with gap junction-mediated communication and ion transport in several other systems, and compared its effects with those of its inactive derivative, 4-O-methyl-TPA. We found that the communication between mouse BALB/c 3T3 cells and human diploid fibroblasts was reduced in the presence of TPA but not O-methyl-TPA and that this inhibition was not obscured by small but measurable influences of TPA on steady-state content and transport of 86Rb+. We confirmed these findings using an autoradiographic assay for transfer of uridine derivatives among cells in contact. We also showed that retinoic acid had no effect on communication in the ion transfer assay but that furosemide, an inhibitor of Na(+)-K(+)-2Cl- cotransport, stimulated ion transfer dramatically both in the presence and absence of TPA. These results indicate both the promise and the limitations of the potassium transfer assay for identifying potential modulators of gap junction-mediated cell communication.
Ledbetter
M L
ML
Department of Biology, College of the Holy Cross, Worcester, Massachusetts 01610.
Medrek
P L
PL
Spinney
B M
BM
D016475:3T3 Cells; D000818:Animals; D001692:Biological Transport; D002450:Cell Communication / Q000187:drug effects*; D002478:Cells, Cultured; D005665:Furosemide / Q000494:pharmacology*; D006801:Humans; D007231:Infant, Newborn; D051379:Mice; D010042:Ouabain / Q000494:pharmacology; D011188:Potassium / Q000378:metabolism*; D012413:Rubidium / Q000378:metabolism; D013755:Tetradecanoylphorbol Acetate / Q000031:analogs & derivatives / Q000494:pharmacology*; D014212:Tretinoin / Q000494:pharmacology*
D014212:Tretinoin; C020922:4-O-methyl-12-O-tetradecanoylphorbol 13-acetate; D010042:Ouabain; D005665:Furosemide; D012413:Rubidium; D013755:Tetradecanoylphorbol Acetate; D011188:Potassium