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1 Department of Physiology and Biophysics, Case Western Reserve University, Cleveland, Ohio 44106; and 2 CJF, Institut National de la Santé et de la Recherche Médicale 94-04, Faculté de Médecine, Université de Nantes, F-44035 Nantes, France
Several
secretagogues induce mucin secretion in epithelial monolayers, as
determined by measuring released granule contents. To assess whether
different agonists act on the same granule pool, capacitance changes in
intact monolayers of the goblet cell line HT29-Cl.16E were measured by
a novel impedance method. Apical ATP (purinergic agonist) and
basolateral carbachol (cholinergic agonist) induce rapid exocytosis
with maximal capacitance changes within 3 min. The maximal levels of
exocytosis that can be induced by optimal concentrations of either
agonist are the same and produce a 30-40% increase in total
monolayer capacitance. When ATP and carbachol are applied
simultaneously, the magnitude of exocytosis is unchanged from the
single-secretagogue level. The recovery of capacitance to baseline
(endocytosis) is significantly faster after ATP stimulation than after
carbachol stimulation. When ATP and carbachol are applied sequentially
at doses that give maximal exocytosis, the magnitude of the capacitance
increase produced by the second secretagogue is less than or equal to
that of the capacitance decrease during the recovery period. Together,
these data suggest that purinergic and cholinergic agonists act on the same granule pool.
mucin secretion; electrophysiology; chloride secretion; membrane capacitance; epithelia
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