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Detection of Mitochondrial Diseases pp 101–113Cite as

Oxidative phosphorylation in myocardial mitochondria ‘in situ’: a calorimetric study on permeabilized cardiac muscle preparations

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Part of the book series: Developments in Molecular and Cellular Biochemistry ((DMCB,volume 21))

Abstract

A novel flow calorimetric technique was developed to study the energy turnover of myocardial mitochondria. Cylindrical strands of cardiac muscle (trabeculae) weighing 100–500 μg were isolated from guinea-pig heart and mounted in a tubular recording chamber which was continuously perfused with physiological salt solution at 37° C. The temperature difference between the upstream and the downstream side of the chamber, which is proportional to the rate of heat production of the trabecula, was measured at high resolution. In this way the rate of energy expenditure of isolated cardiac muscle could be recorded continuously for several hours. When the preparations were superfused with an ‘intracellular’ solution containing 5 mM pyruvate and 2 mM malate as substrates, permeabilization of the sarcolemma with 25 μM digitonin induced a marked increase in the measured heat rate in the presence of 2 mM ADP The major fraction of the ADP sensitive heat production (83%) could be blocked with 400 μM atractyloside, an inhibitor of the adeninenucleotide translocase, and by 600 μM α-cyano-4-hydroxycinnamate, an inhibitor of monocarboxylate/H+ co-transport. The atractyloside sensitive heat production was abolished in anoxic solution. These results suggest that the atractyloside-sensitive heat production (21.8 ± 3.5 mW cm-3 of tissue) was attributable to oxidative phosphorylation. The mitochondria apparently remained intact after treatment with digitonin, since application of the uncoupler 2,4-dinitrophenol (DNP) produced a very large increase in heat rate. A minor fraction of the heat rate induced by ADP in permeabilized cardiac muscle preparations (17%) was not sensitive to atractyloside. This component was also seen before application of digitonin and was probably related to ectonucleotidases. In conclusion, our calorimetric technique allows investigation of the energy metabolism of myocardial mitochondria ‘in situ’ i.e. without destroying the microarchitecture of cardiac muscle cells. (Mol Cell Biochem 174: 101–113, 1997)

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Authors and Affiliations

  1. Institut für Normale und Pathologische Physiologie, Universität Marburg, Marburg, Germany

    Detlef Köhnke & Jürgen Daut

  2. Physiologisches Institut der Technischen Universität München, München, Germany

    Marion Schramm

Authors
  1. Detlef Köhnke
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  2. Marion Schramm
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  3. Jürgen Daut
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Editors and Affiliations

  1. Martin Luther Universität Halle-Wittenberg, Neurologische Klinik und Poliklinik, Halle / Saale, Germany

    Frank Norbert Gellerich  & Stephan Zierz  & 

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Köhnke, D., Schramm, M., Daut, J. (1997). Oxidative phosphorylation in myocardial mitochondria ‘in situ’: a calorimetric study on permeabilized cardiac muscle preparations. In: Gellerich, F.N., Zierz, S. (eds) Detection of Mitochondrial Diseases. Developments in Molecular and Cellular Biochemistry, vol 21. Springer, Boston, MA. https://doi.org/10.1007/978-1-4615-6111-8_16

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  • DOI: https://doi.org/10.1007/978-1-4615-6111-8_16

  • Publisher Name: Springer, Boston, MA

  • Print ISBN: 978-1-4613-7800-6

  • Online ISBN: 978-1-4615-6111-8

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