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The role of cardiac mitochondria in the regulation of intracellular calcium during ischemia and reperfusion: X-ray microanalysis using freeze-dried sections

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Summary

The calcium concentration in papillary muscles was measured by X-ray microanalysis in order to clarify the role played by mitochondria in intracellular calcium regulation during ischemia and reperfusion. Rat hearts perfused by the Langendorff method were rapidly frozen prior to and during ischemia, as well as following reperfusion. Sections prepared by cryoultramicrotomy were freeze-dried, carbon-coated, and analyzed in an electron microscope. A new freeze-drying procedure was developed, in which the ultrastructure was well-preserved, with sarcomeres, triads, and mitochondria easily recognized. Calcium accumulation into the mitochondria occurred during 30-min ischemia (29.7 ± 17.0 mmol/kg dry weight) and increased further after 15-min reperfusion (157.1 ± 104.5), the calcium concentration decreased after 60-min reperfusion (58.1 ± 29.0). However, the calcium concentration in the cytosol did not change significantly. It is thought that mitochondrial calcium accumulation is reversible, to a certain degree, and that the mitochondria play a part in intracellular calcium regulation in pathological states.

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References

  1. Horikawa Y, Kaneko N, Uchida T, Hosoda S, Teraoka K, Kawakami M, Takeishi M, Matsuda R (1989) Change of subcellular calcium induced by ischemia: Electron probe X-ray microanalysis. Eur Heart J 10 [Suppl]:268

    Google Scholar 

  2. Poole-Wilson PA, Harding DP, Bourdillon V, Tomes MA (1984) Calcium out of control. J Mol Cell Cardiol 16:175–187

    Google Scholar 

  3. Stone D, Darley-Usmar V, Smith DR, O'Leary V (1989) Hypoxia-reoxygenation-induced increase in cellular Ca2+ in myocytes and perfused hearts: The role of mitochondria. J Mol Cell Cardiol 21:963–973

    Google Scholar 

  4. Hearse DJ, Humphrey SM, Bullock GR (1978) The oxygen paradox and the calcium paradox: Two facets of the same problem? J Mol Cell Cardiol 10:641–668

    Google Scholar 

  5. Dhalla NS, Pierce GN, Panagia V, Signal PK, Beamish RE (1982) Calcium movements in relation to heart function. Basic Res Cardiol 77:117–139

    Google Scholar 

  6. Carafoli E (1982) The transport of calcium across the inner membrane of mitochondria. In: Carafoli E (ed) Membrane transport of calcium. Academic, London, pp 109–139

    Google Scholar 

  7. Scarpa A, Graziotti P (1973) Mechanisms for intracellular calcium regulation in heart. 1. Stopped-flow measurement of Ca2+ uptake by cardiac mitochondria. J Gen Physiol 62(6):756–772

    Google Scholar 

  8. Lehninger AL, Fiskum G, Vercesi A, Tew W (1981) Ca2+ transport by mitochondria: A survey. In: Bronnert F, Peterlik M (eds) Calcium and phosphate transport across biomembranes. Academic, New York, pp 73–78

    Google Scholar 

  9. Miller DJ (1985) The mitochondria and cellular calcium. Nature 313:638

    Google Scholar 

  10. Somlyo AV, Shuman H, Somlyo AP (1977) Elemental distribution in striated muscle and the effects of hypertonicity: Electron probe analysis of cryo sections. J Cell Biol 74:828–857

    Google Scholar 

  11. Somlyo AV, Sileox J, Somlyo AP (1975) Electron probe analysis and cryo ultramicrotomy of cardiac muscle: Mitochondrial granules. In: Baily GW (ed) Proc 33rd Ann EMSA meet. San Francisco Press, San Francisco, p 532

    Google Scholar 

  12. Buja LM, Burton KP, Hagler HK, Willerson JT (1983) Quantitative X-ray microanalysis of the elemental composition of individual myocytes in hypoxic rabbit myocardium. Circulation 68:872–882

    Google Scholar 

  13. Hagler HK, Sherwin L, Buja LM (1979) Effect of different methods of tissue preparation on mitochondrial inclusions of ischemic and infarcted canine myocardium: Transmission and analytic electron microscopic study. Lab Invest 40:529–544

    Google Scholar 

  14. Hall TA (1971) The microprobe assay of chemical elements. In: Oster G (ed) Physical techniques in biological research 1A. Academic, New York, pp 157–275

    Google Scholar 

  15. Shuman H, Somlyo AV, Somlyo AP (1976) Quantitative electron probe microanalysis of biological thin sections: Methods and validity. Ultramicroscopy 1:317–339

    Google Scholar 

  16. Hon-Chi Lee, Mohabir R, Smith N, Franz MR, Clusin WT (1988) Effect of ischemia on calcium-dependent fluorescence transients in rabbit hearts containing indo-1. Circulation 78:1047–1059

    Google Scholar 

  17. Allshire A, Piper HM, Roy Cuthbertson KS, Cobbold PH (1987) Cytosolic free Ca2+ in single rat heart cells during anoxia and reoxygenation. Biochem J 244:381–385

    Google Scholar 

  18. Miyata H, Silverman HS, Sollott SJ, Lakatta EG, Stern MD, Hansford RG (1991) Measurement of mitochondrial free Ca2+ concentration in living single rat cardiac myocytes. Am J Physiol 261:H1123–H1134

    Google Scholar 

  19. Allen SP, Darley-Usmar VM, McCormack JG, Stone D (1993) Changes in mitochondrial matrix free calcium in perfused rat hearts subjected to hypoxia-reoxygenation. J Mol Cell Cardiol 25:949–958

    Google Scholar 

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Horikawa, Y., Kaneko, N. & Hosoda, S. The role of cardiac mitochondria in the regulation of intracellular calcium during ischemia and reperfusion: X-ray microanalysis using freeze-dried sections. Heart Vessels 10, 1–6 (1995). https://doi.org/10.1007/BF01745071

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  • DOI: https://doi.org/10.1007/BF01745071

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