Abstract
The measurement of oxygen consumption in small tissue regions has been problematic. The perfusion of myocardium is very heterogeneous,1 and it was desirable to measure whether aerobic metabolic rate paralleled the blood flow heterogeneity. However, no method was available to measure local oxygen consumption in tissue regions of a similar size as used for measurement of local blood flow with labeled microspheres. Our goal was to develop a method to measure local O2 consumption (VO2) in many tissue samples simultaneously. VO2 is proportional to the rate in the tricarboxylic acid (TCA) cycle. Methods to determine the flux in the TCA cycle by measuring the gradual enrichment of glutamate with 13C isotopes with NMR spectroscopy exist.2–6 NMR spectra are measured every few minutes until a steady state has been reached, and the time course of enrichment is analyzed with a model for the isotope distribution in the TCA cycle.2 Local blood flow is measured in many small regions simultaneously, and VO2 too must be assessed in many regions simultaneously, which is not yet feasible with NMR coils in vivo. Our strategy is to freeze tissue sam ples quickly at a prescribed time briefly after starting infusion of 13C-enriched substrate for the TCA cycle (e.g. acetate, pyruvate, lactate) and to analyze the 13C NMR multiplets of glutamate in extracts of the samples. We will show below that measurement of O2 consumption is feasible in this way, and test the method in isolated perfused rabbit hearts.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Preview
Unable to display preview. Download preview PDF.
References
Bassingthwaighte JB, King RB, Roger SA. Fractal nature of regional myocardial flow heterogeneity. Circ Res 1989; 65: 578–590.
Chance EM, Seeholzer, SH, Kobayashi K, Williamson JR. Mathematical analysis of isotope labeling in the citric acid cycle with applications to 13C NMR studies in perfused rat hearts. J Biol Chem 1983; 258: 13785–13794.
Weiss RG, Gloth ST, Kalil-Filho R, Chacko VP, Stern MD, Gerstenblith G. Indexing tricarboxylic acid cycle flux in intact hearts by carbon-13 nuclear magnetic resonance. Circ Res 1992; 70: 392–408.
Chatham JC, Forder JR, Glickson JD, Chance EM. Calculation of absolute metabolic flux and the elucidation of pathways of glutamate labeling in perfused rat heart by I3C NMR spectroscopy and nonlinear least squares analysis. J Biol Chem 1995; 270: 7999–8008.
Lewandowski ED. Nuclear magnetic resonance evaluation of metabolic and respiratory support of work load in intact rabbit hearts. Circ Res 1992; 70: 576–582.
Yu X, White LT, Doumen C, Damico LA, LaNoue KF, Alpert NM, Lewandowski ED. Kinetic analysis of dynamic I3C NMR spectra: metabolic flux, regulation, and eompartmentation in hearts. Biophys J 1995; 69:2090–2102.
Malloy CR, Sherry AD, Jeffrey FMH. Analysis of tricarboxylic acid cycle of the heart using I3C isotope isomers. Am J Physiol 1990; 259: H987–H995.
Malloy CR, Thompson JR, Jeffrey FMH, Sherry AD. Contribution of exogenous substrates to acetyl coen-zyme A: measurement by 13C NMR under non-steady-state conditions. Biochemistry 1990; 29: 6756–6761.
Van Beek JHGM, Bussemaker J, Westerhof N. Measurement of local oxygen consumption in small frozen tissue samples with a new method shows a higher metabolic rate in subendocardium than in subepicardium in isolated rabbit heart. J Physiol 1996; P.
Van Beek JHGM, Bussemaker J, Barends JPF, Westerhof N. A 13C-NMR technique to determine absolute oxygen consumption in quickly frozen small myocardial samples. FASEB J 1996; 10:A325.
Van Beek JHGM, Csont T, Bussemaker J, Barends JPF. Measuring local myocardial O2 consumption in many samples. Ann Biomed Eng 1996; 24:S–19.
Van Beek JHGM, de Kanter FJJ, Bussemaker J. Measuring subendocardial versus subepicardial O2 consumption in rabbit left ventricle using 13C-NMR spectroscopy. J Moll Cell Cardiol (submitted).
Van Beek JHGM, Westerhof N. Response time of cardiac mitochondrial oxygen consumption to heart rate steps. Am J Physiol 1991; 260: H613–H625.
Knijn A, De Beer R, Van Ormondt D. Frequency-selective quantification in the time domain. J Magn Reson 1992; 97: 444–450.
Van den Bogaart A, Ala-Korpela M, Jokisaari J, Griffiths JR. Time and frequency domain analysis of NMR data compared: an application to ID 1H spectra of lipoproteins. Magn Reson Med 1994; 31:347–358.
Randle PJ, England PJ, Denton RM. Control of the tricarboxylate cycle and its interactions with glycolysis during acetate utilization in rat heart. Biochem J 117: 677–695.
Li Z, Yipintsoi T, Caldwell JH, Zuurbier CJ, Krohn KA, Link JM, Bassingthwaighte JB. In vivo measurement of regional myocardial oxygen utilization with inhaled l5O-oxygen and positron emission tomography. Ann Biomed Eng 1996; 24: S–32.
Van Beek JHGM. Is local metabolism the basis of the fractal vascular structure in the heart? Int J Microcirc (In press) 9 pp
Bussemaker J, Groeneveld ABJ, Teerlink T, Hennekes M, Westerhof N, Van Beek JHGM. Low-and high blood flow regions in the normal pig heart are equally vulnerable to ischaemia during partial coronary stenosis. Eur J Physiol (Pflügers Arch) 1997; 434:785–794.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 1998 Springer Science+Business Media New York
About this chapter
Cite this chapter
van Beek, J.H.G.M., Csont, T., de Kanter, F.J.J., Bussemaker, J. (1998). Simple Model Analysis of 13C NMR Spectra to Measure Oxygen Consumption Using Frozen Tissue Samples. In: Hudetz, A.G., Bruley, D.F. (eds) Oxygen Transport to Tissue XX. Advances in Experimental Medicine and Biology, vol 454. Springer, Boston, MA. https://doi.org/10.1007/978-1-4615-4863-8_58
Download citation
DOI: https://doi.org/10.1007/978-1-4615-4863-8_58
Publisher Name: Springer, Boston, MA
Print ISBN: 978-1-4613-7206-6
Online ISBN: 978-1-4615-4863-8
eBook Packages: Springer Book Archive