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Am J Physiol Heart Circ Physiol 279: H1179-H1184, 2000;
0363-6135/00 $5.00
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Vol. 279, Issue 3, H1179-H1184, September 2000

Altered cardiac Na+/H+ exchange in phospholipase D-treated sarcolemmal vesicles

Danny P. Goel1, Alba Vecchini3, Vincenzo Panagia2, and Grant N. Pierce1

1 Division of Stroke and Vascular Disease and 2 Institute of Cardiovascular Sciences, St. Boniface General Hospital Research Centre, and Departments of Physiology and Human Anatomy and Cell Science, University of Manitoba, Winnipeg, Canada R2H 2A6; and 3 Institute of Biochemistry and Medical Chemistry, University of Perugia, Italy 06100

Cardiac sarcolemmal Na+/H+ exchange is critical for the regulation of intracellular pH, and its activity contributes to ischemia-reperfusion injury. It has been suggested that the membrane phospholipid environment does not modulate Na+/H+ exchange. The present study was carried out to determine the effects on Na+/H+ exchange of modifying the endogenous membrane phospholipids through the addition of exogenous phospholipase D. Incubation of 0.825 U of phospholipase D with 1 mg of porcine cardiac sarcolemmal vesicles hydrolyzed 34 ± 2% of the sarcolemmal phosphatidylcholine and increased phosphatidic acid 10.2 ± 0.5-fold. Treatment of vesicles with phospholipase D resulted in a 46 ± 2% inhibition of Na+/H+ exchange. Na+/H+ exchange was measured as a function of reaction time, extravesicular pH, and extravesicular Na+. All of these parameters of Na+/H+ exchange were inhibited following phospholipase D treatment compared with untreated controls. Passive efflux of Na+ was unaffected. Treatment of sarcolemmal vesicles with phospholipase C had no effect on Na+/H+ exchange. We conclude that phospholipase D-induced changes in the cardiac sarcolemmal membrane phospholipid environment alter Na+/H+ exchange.

phospholipids; phospholipases; Na+/H+ exchanger isoform 1; ischemia; preconditioning; membrane


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