Тhapsigargin- sensitive and insensitiveintracellular са[sup]2+[/sup] stores in ratsubmandibular salivary gland acinarcells
О.V. Коpach, І.А. Kruglikov,N.V. Voitenko, N.V. Fedirko
Ivan Franko National University of LvivA.A. Bogomolets Institute of Physiology National Academy ofSciences of Ukraine, Kyiv
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Abstract
Acinar cells of rat submandibular salivary gland are
characterized by heterogeneity of intracellular Ca 2+ stores. In
the present work we have studied this heterogeneity using
Arsenazo III dye to measure a cellular total calcium content
and Fura-2/AM, to determine free cytosolic calcium
concentration ([Ca 2+]i). We have found that the amount of Ca 2+
released by inhibition of Ca 2+ ATPase of the ER with
thapsigargin comprises ~ 30 % of total ER calcium. This result
was obtained in experiments on both intact and permeabilized
acinar cells. We have also shown that both Ca2+ ATPase inhibi-
tion with thapsigargin and emptying the stores with acetyl-
choline (ACh) leaded to activation of store-operated Ca 2+ influx
(an increase in total calcium content of ~ 14 %). In
permeabilized cells application of ACh after preincubation
with thapsigargin leaded to a further decrease in total cellular
calcium content (~ 38%). At the same time in intact cells it
resulted in generation of [Ca 2+]i transients with gradually de-
creasing amplitudes. Thus, ACh is capable of producing an
additional release of Ca 2+ from thapsigargin-insensitive stores.
This additional release is IP3 -dependent since it was completely
blocked by heparin. We conclude that in acinar cells of rat
submandibular gland thapsigargin-sensitive and thapsigargin-
insensitive Ca2+ stores could exist.
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