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secretion in T84
cells: membrane target(s) of inhibition is agonist
specific
Department of Medicine, University of California, San Diego, School of Medicine, San Diego, California 92103
Previous studies have indicated that
Ca2+-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
Ca2+-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
Ca2+-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.
chloride channels; potassium channels; 3-phosphorylated lipids; calcium
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