Inhibition of Ca(2+)-dependent Cl- secretion in T84 cells: membrane target(s) of inhibition is agonist specific.
Previous studies have indicated that Ca(2+)-dependent Cl- secretion across monolayers of T84 epithelial cells is subject to a variety of negative influences that serve to limit the overall extent of secretion. However, the downstream membrane target(s) of these inhibitory influences had not been elucidated. In this study, nuclide efflux techniques were used to determine whether inhibition of Ca(2+)-dependent Cl- secretion induced by carbachol, inositol 3,4,5,6-tetrakisphosphate, epidermal growth factor, or insulin reflected actions at an apical Cl- conductance, a basolateral K+ conductance, or both. Pretreatment of T84 cell monolayers with carbachol or a cell-permeant analog of inositol 3,4,5,6-tetrakisphosphate reduced the ability of subsequently added thapsigargin to stimulate apical 125I-, but not basolateral 86Rb+, efflux. These data suggested an effect on an apical Cl- channel. Conversely, epidermal growth factor reduced Ca(2+)-stimulated 86Rb+ but not 125I- efflux, suggesting an effect of the growth factor on a K+ channel. Finally, insulin inhibited 125I- and 86Rb+ effluxes. Thus effects of agents that inhibit transepithelial Cl- secretion are also manifest at the level of transmembrane transport pathways. However, the precise nature of the membrane conductances targeted are agonist specific.
Barrett
K E
KE
Department of Medicine, University of California, San Diego, School of Medicine 92103, USA. kbarrett@ucsd.edu
Smitham
J
J
Traynor-Kaplan
A
A
Uribe
J M
JM
D002118:Calcium / Q000502:physiology*; D002217:Carbachol / Q000494:pharmacology; D002460:Cell Line; D002712:Chlorides / Q000378:metabolism*; D004815:Epidermal Growth Factor / Q000494:pharmacology; D006632:Histamine / Q000494:pharmacology; D006801:Humans; D007328:Insulin / Q000494:pharmacology; D007413:Intestinal Mucosa / Q000166:cytology / Q000378:metabolism*; D007455:Iodine / Q000037:antagonists & inhibitors / Q000378:metabolism; D012413:Rubidium / Q000037:antagonists & inhibitors / Q000378:metabolism; D013379:Substrate Specificity; D019284:Thapsigargin / Q000494:pharmacology
D002712:Chlorides; D007328:Insulin; D004815:Epidermal Growth Factor; D019284:Thapsigargin; D006632:Histamine; D002217:Carbachol; D007455:Iodine; D012413:Rubidium; D002118:Calcium