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Am J Physiol Heart Circ Physiol 277: H1630-H1640, 1999;
0363-6135/99 $5.00
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Vol. 277, Issue 4, H1630-H1640, October 1999

SPECIAL COMMUNICATION
A 13C NMR double-labeling method to quantitate local myocardial O2 consumption using frozen tissue samples

Johannes H. G. M. van Beek1, Harald G. J. van Mil1,3, Richard B. King4, Frans J. J. de Kanter2, David J. C. Alders1, and Joli Bussemaker1

Laboratory for Physiology, 1 Institute for Cardiovascular Research and 2 Division of Chemistry, Vrije Universiteit, 1081 BT Amsterdam; and 3 Theory of Complex Fluids Group, Faculty of Applied Science, Delft University of Technology, 2628 BC Delft, The Netherlands; and 4 National Simulation Resource for Circulatory Mass Transport and Exchange, Center for Bioengineering, University of Washington, Seattle, Washington 98195

Measurement of local myocardial O2 consumption (VO2) has been problematic but is needed to investigate the heterogeneity of aerobic metabolism. The goal of the present investigation was to develop a method to measure local VO2 using small frozen myocardial samples, suitable for determining VO2 profiles. In 26 isolated rabbit hearts, 1.5 mmol/l [2-13C]acetate was infused for 4 min, followed by 1.5 min of [1,2-13C]acetate. The left ventricular (LV) free wall was then quickly frozen. High-resolution 13C-NMR spectra were measured from extracts taken from 2- to 3-mm thick transmural layer samples. The multiplet intensities of glutamate were analyzed with a computer model allowing simultaneous estimation of the absolute flux through the tricarboxylic acid cycle and the fractional contribution of acetate to acetyl CoA formation from which local VO2 was calculated. The 13C-derived VO2 in the LV free wall was linearly related to "gold standard" VO2 from coronary venous O2 electrode measurements in the same region (r = 0.932, n = 22, P < 0.0001, slope 1.05) for control and lowered metabolic rates. The ratio of subendocardial to subepicardial VO2 was 1.52 ± 0.19 (SE, significantly >1, P < 0.025). Local myocardial VO2 can now be quantitated with this new 13C method to determine profiles of aerobic energy metabolism.

myocardial metabolism; heterogeneity of metabolism; tricarboxylic acid cycle; metabolism-perfusion matching; stable isotopes


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