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Am J Physiol Heart Circ Physiol 287: H889-H895, 2004. First published March 25, 2004; doi:10.1152/ajpheart.00976.2003
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A 13C isotopomer kinetic analysis of cardiac metabolism: influence of altered cytosolic redox and [Ca2+]o

Rui A. Carvalho,1 Tiago B. Rodrigues,1 Piyu Zhao,2 F. Mark H. Jeffrey,3 Craig R. Malloy,3,4 and A. Dean Sherry2,3

1Department of Biochemistry and Center for Neurosciences and Cell Biology, Faculty of Sciences and Technology, University of Coimbra, Apartado 3126, 3001-401 Coimbra, Portugal; 2Department of Chemistry, University of Texas at Dallas, Richardson 75083-0688; 3Department of Radiology, Mary Nell and Ralph B. Rogers Magnetic Resonance Center, University of Texas Southwestern Medical Center; and 4Department of Internal Medicine, University of Texas Southwestern Medical Center, Dallas 75235-9085; and Department of Veterans Affairs Medical Center, Dallas, Texas 75216

Submitted 14 October 2003 ; accepted in final form 18 March 2004

Rat hearts were perfused with mixtures of [3-13C]pyruvate and [3-13C]lactate (to alter cytosolic redox) at low (0.5 mM) or high (2.5 mM) Ca2+ concentrations to alter contractility. Hearts were frozen at various times after exposure to these substrates, were extracted, and were then analyzed by 13C NMR spectroscopy. The time-dependent multiplets observed in the 13C NMR resonances of glutamate in all hearts and in malate and aspartate in hearts perfused with high-pyruvate/low-lactate concentrations were analyzed using a kinetic model of the tricarboxylic acid (TCA) cycle. The analysis showed that TCA cycle flux (VTCA) and exchange flux (VX) that involved cycle intermediates were both sensitive to cell redox and altered Ca2+ concentration, and the ratio of these fluxes (VX/VTCA) varied >10-fold.

13C nuclear magnetic resonance; tricarboxylic acid cycle; intermediary metabolism; malate-aspartate shuttle; modeling; extracellular calcium concentration


Address for reprint requests and other correspondence: A. D. Sherry, Mary Nell and Ralph B. Rogers Magnetic Resonance Center, 5801 Forest Park Rd., Dallas, TX 75235-9085 (E-mail: Dean.Sherry{at}UTSouthwestern.edu).






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