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Modulation of two functionally distinct Ca 2+ stores in astrocytes: Role of the plasmalemmal Na/Ca exchanger

Modulation of two functionally distinct Ca 2+ stores in astrocytes: Role of the plasmalemmal... 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) Ca2+‐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 Ca2+‐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, Ca2+‐ (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. © 1996 Wiley‐Liss, Inc. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Glia Wiley

Modulation of two functionally distinct Ca 2+ stores in astrocytes: Role of the plasmalemmal Na/Ca exchanger

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References (47)

Publisher
Wiley
Copyright
Copyright © 1996 Wiley‐Liss, Inc.
ISSN
0894-1491
eISSN
1098-1136
DOI
10.1002/(SICI)1098-1136(199604)16:4<296::AID-GLIA2>3.0.CO;2-Z
Publisher site
See Article on Publisher Site

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) Ca2+‐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 Ca2+‐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, Ca2+‐ (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. © 1996 Wiley‐Liss, Inc.

Journal

GliaWiley

Published: Apr 1, 1996

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