Abstract:
:Mechanisms that regulate the amount of releasable Ca2+ in intracellular stores of cultured mouse astrocytes were investigated using digital imaging of fura-2 loaded cells. At rest, the cytoplasmic Ca2+ concentration, [Ca2+]cyt, was about 110 nM. In the absence of extracellular Ca2+, cyclopiazonic acid (CPA), an inhibitor of the endoplasmic reticulum (ER) Ca(2+)-ATPase, induced a transient, four-fold increase in [Ca2+]cyt due to the release of Ca2+ from inositol triphosphate (IP3) sensitive stores. Caffeine (CAF), which releases Ca2+ from Ca(2+)-sensitive stores, induced a two-fold increase in [Ca2+]cyt. The CPA- and CAF-sensitive stores could be released independently. Changes in the amplitudes of the Ca2+ transients were taken as a measure of changes in store content. Removal of extracellular Na+ or addition of ouabain, which inhibit Ca2+ extrusion and promote Ca2+ entry across the plasmalemma via the Na/Ca exchanger, caused minimal increases in resting [Ca2+]cyt but greatly potentiated both CPA- and CAF-induced Ca2+ transients. The amount of Ca2+ releasable from the IP3(CPA) sensitive store was directly proportional to cytosolic Na+ concentration (i.e., inversely proportional to the transmembrane Na+ electrochemical gradient). Under these reduced Na+ gradient conditions, little, if any, Ca2+ destined for the ER stores enters the cells through voltage-dependent Ca2+ channels. These results demonstrate that mouse astrocytes contain two distinct ER Ca2+ stores, the larger, IP3- (CPA-) sensitive, and the smaller, Ca(2+)- (CAF-) sensitive. The Ca2+ content of both ER stores can be regulated by the Na/Ca exchanger. Thus, the magnitude of cellular responses to signals that are mediated by Ca2+ release induced by the two second messengers, IP3 and Ca2+, can be modulated by factors that affect the net transport of Ca2+ across the plasmalemma.
journal_name
Gliajournal_title
Gliaauthors
Golovina VA,Bambrick LL,Yarowsky PJ,Krueger BK,Blaustein MPdoi
10.1002/(SICI)1098-1136(199604)16:4<296::AID-GLIA2subject
Has Abstractpub_date
1996-04-01 00:00:00pages
296-305issue
4eissn
0894-1491issn
1098-1136pii
10.1002/(SICI)1098-1136(199604)16:4<296::AID-GLIA2journal_volume
16pub_type
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