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Am J Physiol Cell Physiol 276: C1091-C1099, 1999;
0363-6143/99 $5.00
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Vol. 276, Issue 5, C1091-C1099, May 1999

Mechanisms of Na+-K+ pump regulation in cardiac myocytes during hyposmolar swelling

N. L. Bewick1, C. Fernandes1,2, A. D. Pitt1, H. H. Rasmussen1,2, and D. W. Whalley1,2

1 Cardiology Department, Royal North Shore Hospital, St. Leonards, New South Wales 2065; and 2 University of Sydney, Sydney, New South Wales 2006, Australia

We have previously demonstrated that the sarcolemmal Na+-K+ pump current (Ip) in cardiac myocytes is stimulated by cell swelling induced by exposure to hyposmolar solutions. However, the underlying mechanism has not been examined. Because cell swelling activates stretch-sensitive ion channels and intracellular messenger pathways, we examined their role in mediating Ip stimulation during exposure of rabbit ventricular myocytes to a hyposmolar solution. Ip was measured by the whole cell patch-clamp technique. Swelling-induced pump stimulation altered the voltage dependence of Ip. Pump stimulation persisted in the absence of extracellular Na+ and under conditions designed to minimize changes in intracellular Ca2+, excluding an indirect influence on Ip mediated via fluxes through stretch-activated channels. Pump stimulation was protein kinase C independent. The tyrosine kinase inhibitor tyrphostin A25, the phosphatidylinositol 3-kinase inhibitor LY-294002, and the protein phosphatase-1 and -2A inhibitor okadaic acid abolished Ip stimulation. Our findings suggest that swelling-induced pump stimulation involves the activation of tyrosine kinase, phosphatidylinositol 3-kinase, and a serine/threonine protein phosphatase. Activation of this messenger cascade may cause activation by the dephosphorylation of pump units.

osmolarity; tyrosine kinase; ion transport; phosphatases


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