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
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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