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Am J Physiol Heart Circ Physiol 280: H1989-H1995, 2001;
0363-6135/01 $5.00
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Vol. 280, Issue 5, H1989-H1995, May 2001

Lack of muscarinic regulation of Ca2+ channels in Gi2alpha gene knockout mouse hearts

Fuhua Chen1, Karsten Spicher2, Meisheng Jiang2, Lutz Birnbaumer2, and Glenn T. Wetzel1

1 Department of Pediatrics and 2 Departments of Anesthesiology and Biological Chemistry, University of California School of Medicine, Los Angeles, California 90095

The purpose of the present study was to examine the role of Gi2alpha in Ca2+ channel regulation using Gi2alpha gene knockout mouse ventricular myocytes. The whole cell voltage-clamp technique was used to study the effects of the muscarinic agonist carbachol (CCh) and the beta -adrenergic agonist isoproterenol (Iso) on cardiac L-type Ca2+ currents in both 129Sv wild-type (WT) and Gi2alpha gene knockout (Gi2alpha -/-) mice. Perfusion with CCh significantly inhibited the Ca2+ current in WT cells, and this effect was reversed by adding atropine to the CCh-containing solution. In contrast, CCh did not affect Ca2+ currents in Gi2alpha -/- ventricular myocytes. Addition of CCh to Iso-containing solutions attenuated the Iso-stimulated Ca2+ current in WT cardiomyocytes but not in Gi2alpha -/- cells. These findings demonstrate that, whereas the Iso-Gsalpha signal pathway is intact in Gi2alpha gene knockout mouse hearts, these cells lack the inhibitory regulation of Ca2+ channels by CCh. Therefore, Gi2alpha is necessary for the muscarinic regulation of Ca2+ channels in the mouse heart. Further studies are needed to delineate the possible interaction of Gi and other cell signaling proteins and to clarify the level of interaction of G protein-coupled regulation of L-type Ca2+ current in the heart.

Ca2+ current regulation; gene inactivation; signal transduction


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