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Originally published In Press as doi:10.1074/jbc.M009179200 on December 4, 2000

J. Biol. Chem., Vol. 276, Issue 8, 5650-5658, February 23, 2001
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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*

Thomas Meyer, Marie-Cécile Wellner-Kienitz, Anke Biewald, Kirsten Bender, Andreas Eickel, and Lutz PottDagger

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, alpha -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 GTPgamma S (guanosine 5'-3-O-(thio)triphosphate), causing stable activation of IK(ACh), inhibition via alpha -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 GTPgamma S-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 alpha 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.

Dagger 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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