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J. Biol. Chem., Vol. 277, Issue 29, 26479-26485, July 19, 2002
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From the Medical Research Council Secretory Control Research Group,
The Physiological Laboratory, University of Liverpool,
Liverpool L69 3BX, United Kingdom
We have studied the Ca2+ leak
pathways in the endoplasmic reticulum of pancreatic acinar cells by
directly measuring Ca2+ in the endoplasmic reticulum
([Ca2+]ER). Cytosolic
Ca2+ ([Ca2+]C) was
clamped to the resting level by a BAPTA-Ca2+ mixture.
Administration of cholecystokinin within the physiological concentration range caused a graded decrease of
[Ca2+]ER, and the rate of
Ca2+ release generated by 10 pM cholecystokinin
is at least 3× as fast as the basal Ca2+ leak revealed by
inhibition of the endoplasmic reticulum Ca2+-ATPase.
Acetylcholine also evokes a dose-dependent decrease of [Ca2+]ER, with an
EC50 of 0.98 ± 0.06 µM. Inhibition of
receptors for inositol 1,4,5-trisphosphate (IP3) by heparin
or flunarizine blocks the effect of acetylcholine but only partly
blocks the effect of cholecystokinin. 8-NH2 cyclic
ADP-ribose (20 µM) inhibits the action of
cholecystokinin, but not of acetylcholine. The basal Ca2+ leak from the endoplasmic reticulum is not blocked by
antagonists of the IP3 receptor, the ryanodine receptor, or
the receptor for nicotinic acid adenine dinucleotide phosphate.
However, treatment with puromycin (0.1-1 mM) to
remove nascent polypeptides from ribosomes increases Ca2+
leak from the endoplasmic reticulum by a mechanism independent of the
endoplasmic reticulum Ca2+ pumps and of the receptors for
IP3 or ryanodine.
Basal and Physiological Ca2+ Leak from
the Endoplasmic Reticulum of Pancreatic Acinar Cells
SECOND MESSENGER-ACTIVATED CHANNELS AND TRANSLOCONS*
§,¶,
*
This work was supported by a Medical Research Council
program grant.The costs of publication of this
article were defrayed in part by the
payment of page charges. The article
must therefore be hereby marked
"advertisement" in
accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
These authors made similar contributions to the work.
§
To whom correspondence may be addressed: The Physiological
Laboratory, Crown St., University of Liverpool, Liverpool L69 3BX, UK.
Tel.: 44-151-794-5351; Fax: 44-151-794-5327; E-mail: rlomax@liv.ac.uk (to R. L.) and a.tepikin@liv.ac.uk (to A. T.).
¶
Postdoctoral fellow funded by the Consejería de
Educación, Ciencia y Tecnología de la Junta de Extremadura.
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