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Originally published In Press as doi:10.1074/jbc.M006058200 on August 24, 2000

J. Biol. Chem., Vol. 275, Issue 45, 35402-35407, November 10, 2000
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Intracellular Sodium Modulates Mitochondrial Calcium Signaling in Vascular Endothelial Cells*

Marina Sedova and Lothar A. BlatterDagger

From the Loyola University Chicago, Stritch School of Medicine, Department of Physiology, Maywood, Illinois 60153

We have investigated the role of extramitochondrial Na+ for the regulation of mitochondrial Ca2+ concentration ([Ca2+]m) in permeabilized single vascular endothelial cells. [Ca2+]m was measured by loading the cells with the membrane-permeant Ca2+ indicator fluo-3/AM and subsequent removal of cytoplasmic fluo-3 by surface membrane permeabilization with digitonin. An elevation of extramitochondrial Ca2+ resulted in a dose-dependent increase in the rate of Ca2+ accumulation into mitochondria (k0.5 = 3 µM) via the mitochondrial Ca2+ uniporter. In the presence of 10 mM extramitochondrial Na+ ([Na+]em), repetitive application of brief pulses of high Ca2+ (2-10 µM) to simulate cytoplasmic [Ca2+] oscillations caused transient increases of [Ca2+]m characterized by a fast rising phase that was followed by a slow decay. Removal of extramitochondrial Na+ or inhibition of mitochondrial Na+/Ca2+ exchange with clonazepam blocked mitochondrial Ca2+ efflux and resulted in a net accumulation of Ca2+ by the mitochondria. Half-maximal activation of mitochondrial Na+/Ca2+ exchange occurred at [Na+]em = 4.4 mM, which is well within the physiological range of cytoplasmic [Na+]. This study provides evidence that Ca2+ efflux from the mitochondria in vascular endothelial cells occurs solely via Na+/Ca2+ exchange and emphasizes the important role of intracellular Na+ for mitochondrial Ca2+ regulation.

* Financial support was provided by grants from the National Institutes of Health (HL-51941 and HL-62231) and the American Heart Association National Center (Established Investigator Award 95002520).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: Dept. of Physiology, Loyola University Chicago, 2160 S. First Ave., Maywood, IL 60153. Tel.: 708-216-1182; Fax: 708-216-6308; E-mail: LBLATTE@LUMC.EDU.

Copyright © 2000 by The American Society for Biochemistry and Molecular Biology, Inc.
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