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J. Biol. Chem., Vol. 276, Issue 8, 5650-5658, February 23, 2001
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From the Institut für Physiologie, Ruhr-Universität
Bochum, D44780 Bochum, Germany
G protein-gated inwardly rectifier
K+ current in atrial myocytes (IK(ACh))
upon stimulation with acetylcholine (ACh) shows a fast desensitizing
component (t1/2 ~ 5 s). After washout of
ACh, IK(ACh) recovers from fast desensitization within < 30 s. A recent hypothesis suggests that fast desensitization is
caused by depletion of phosphatidylinositol 4,5-bisphosphate
(PtIns(4,5)P2), resulting from costimulation of
phospholipase C (PLC)-coupled M3 receptors
(M3AChR). The effects of stimulating two established
PLC-coupled receptors,
Depletion of Phosphatidylinositol 4,5-Bisphosphate by Activation
of Phospholipase C-coupled Receptors Causes Slow Inhibition but Not
Desensitization of G Protein-gated Inward Rectifier
K+ Current in Atrial Myocytes*
-adrenergic and endothelin (ETA),
on IK(ACh) were studied in rat atrial myocytes. Stimulation
of these receptors caused activation of IK(ACh) and
inhibition of the M2AChR-activated current. In myocytes
loaded with GTP
S (guanosine 5'-3-O-(thio)triphosphate),
causing stable activation of IK(ACh), inhibition via
-agonists and ET-1 was studied in isolation. Stimulation of either
type of receptor under this condition, via Gq/11, caused a
slow inhibition (t1/2~50 s) by about 70%. No
comparable effect on GTPS-activated IK(ACh) was induced
by ACh, suggesting that PLC-coupled M3AChRs are not
functionally expressed in rat myocytes, which was supported by the
finding that M3AChR transcripts were not detected by
reverse transcriptase-polymerase chain reaction in identified atrial
myocytes. Supplementing the pipette solution with
PtIns(4,5)P2 significantly reduced inhibition of
IK(ACh) but had no effect on fast desensitization. From
these data it is concluded that stimulation of PLC-coupled receptors
causes slow inhibition of IK(ACh) by depletion of
PtIns(4,5)P2, whereas fast desensitization of
IK(ACh) is not related to PtIns(4,5)P2
depletion. As muscarinic stimulation by ACh does not exert inhibition
of IK(ACh) comparable to stimulation of 1-
and ETA receptors, expression of functional PLC-coupled
muscarinic receptors in rat atrial myocytes is unlikely.
*
This work was supported by Grant Po 212/9-3 from the
Deutsche Forschungsgemeinschaft.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.
To whom correspondence should be addressed. Tel.:
49-234-322-9200; Fax: 49-234-321-4449; E-mail:
lutz.pott@ruhr-uni-bochum.de.
Copyright © 2001 by The American Society for Biochemistry and Molecular Biology, Inc.
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