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A comparative study of the calcium accumulation by mitochondria and microsomes isolated from the smooth muscle of the guinea-pig taenia coli

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Summary

The calcium uptake by mitochondria and microsomes isolated from the guinea-pig taenia coli was studied at physiological Ca2+ concentrations, buffered by Ca-EGTA mixtures. The Ca accumulation by the mitochondria was measured from the difference between the amount of Ca taken up in the presence and in the absence of a specific mitochondrial inhibitor. The Ca uptake by the microsomes was determined in a solution containing oxalate and a mitochondrial inhibitor. It was calculated from the difference in Ca uptake measured with and without ATP. By using this procedure, the necessity of extensive purification of the isolated fractions was avoided.

The (Ca2+) for half-maximal transport in the mitochondria is 7×10−6 M. At (Ca2+) lower than 2×10−7 M, Ca is taken up in an energy-dependent way.

In the microsomes the apparentK m for Ca is 7×10−7 M. Accumulation is still stimulated by ATP at a (Ca2+) as low as 4×10−8 M.

The results show that the rate of Ca uptake by the cell organelles corresponding to the microsomal vesicles is sufficiently fast to explain the speed of relaxation of the taenia coli. The results also suggest that these cell organelles are more important than the mitochondria in regulating the cytoplasmic Ca concentration.

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References

  1. Batra, S.: The role of mitochondrial calcium uptake in contraction and relaxation of the human myometrium. Biochim. biophys. Acta (Amst.)305, 428–432 (1973)

    Google Scholar 

  2. Batra, S.: The role of mitochondria in the regulation of myoplasmic calcium concentration in smooth muscle. In: Calcium transport in contraction and secretion (E. Carafoli et al., eds.), pp. 87–94. Amsterdam: North Holland Publishing Company 1975

    Google Scholar 

  3. Beaufay, H., Amar-Costesec, A., Feytmans, E., Thines-Sempaux, D., Wibo, M., Robbi, M., Berthet, J.: Analytical study of microsomes and isolated subcellular membranes from rat liver. I. Biochemical methods. J. Cell Biol.61, 188–200 (1974)

    Google Scholar 

  4. Bianchi, C. P.: Pharmacology of excitation-contraction coupling in muscle. Introduction: Statement of the problem. Fed. Proc.28, 1624–1628 (1969)

    Google Scholar 

  5. Carafoli, E.: Mitochondria, Ca2+ transport and the regulation of heart contraction and metabolism. J. Molec. Cell. Cardiol.7, 83–89 (1975)

    Google Scholar 

  6. Carsten, M. E.: Role of calcium binding by sarcoplasmic reticulum in the contraction and relaxation of uterine smooth muscle. J. gen. Physiol.53, 414–426 (1969)

    Google Scholar 

  7. Casteels, R., van Breemen, C.: Active and passive Ca2+ fluxes across cell membranes of the guinea-pig taenia coli. Pflügers Arch.359, 197–207 (1975)

    Google Scholar 

  8. Ford, G. D., Hess, M. L.: Calcium-accumulating properties of subcellular fractions of bovine vascular smooth muscle. Circulat. Res.37, 580–587 (1975)

    Google Scholar 

  9. Gabella, G.: Fine structure of smooth muscle. Phil. Trans. B265, 7–16 (1973)

    Google Scholar 

  10. Gabella, G., Raeymaekers, L.: Effect of collagenase on mechanical activity and fine structure of an intestinal smooth muscle. Cell Tiss. Res.173, 29–44 (1976)

    Google Scholar 

  11. Godt, R. E.: Calcium-activated tension of skinned muscle fibres of the frog. Dependence on magnesium adenosine triphosphate concentration. J. gen. Physiol.63, 722–739 (1974)

    Google Scholar 

  12. Hurwitz, L., Fitzpatrick, D. F., Debbas, G., Landon, E. J.: Localization of calcium pump activity in smooth muscle. Science179, 384–386 (1973)

    Google Scholar 

  13. Lehninger, A. L.: Ca2+ transport by mitochondria and its possible role in the cardiac contraction relaxation cycle. Circulat. Res.35, III 83–90 (1974)

    Google Scholar 

  14. Lowry, O. H., Rosebrough, N. J., Farr, A. L., Randall, R. J.: Protein measurement with the Folin phenol reagent. J. biol. Chem.193, 265–275 (1951)

    Google Scholar 

  15. Ogawa, Y.: The apparent binding constnt of glycolether-diamine-tetraacetic acid for calcium at neutral pH. J. Biochem. (Tokyo)64, 255–257 (1968)

    Google Scholar 

  16. Popescu, L. M., Diculescu, I., Zelck, V., Ionescu, N.: Ultrastructural distribution of calcium in smooth muscle cells of guinea-pig taenia coli. A correlated electron microscopic and quantitative study. Cell Tiss. Res.154, 357–378 (1974)

    Google Scholar 

  17. Portzehl, H., Caldwell, P. C., Rüegg, J. C.: The dependence of contraction and relaxation of muscle fibres from the crab Maia squinado on the internal concentration of free calcium ions. Biochim. biophys. Acta (Amst.)79, 581–591 (1964)

    Google Scholar 

  18. Raeymaekers, L., Wuytack, F., Casteels, R.: Calcium accumulation by mitochondrial and microsomal fractions isolated from the smooth muscle of the guinea-pig taenia coli. Arch. internat. Physiol. Biochim.84, 355–357 (1976)

    Google Scholar 

  19. Raeymaekers, L., Wuytack, F., Casteels, R.: The affinity of mitochondria and microsomes isolated from the guinea-pig taenia coli for calcium. Proc. of the Fifth International Congress of Histochemistry and Cytochemistry, p. 290, Bucharest 1976

  20. Scarpa, A., Graziotti, P.: Mechanisms for intracellular calcium regulation in heart. I. Stopped-flow measurements of Ca++ uptake by cardiac mitochondria. J. gen. Physiol.62, 756–772 (1973)

    Google Scholar 

  21. Sobieszek, A., Bremel, R. D.: Preparation and properties of vertebrate smooth muscle myofibrils and actomyosin. Europ. J. Biochem.55, 49–60 (1975)

    Google Scholar 

  22. Somlyo, A. P., Somylo, A. V., Devine, C. E., Peters, P. D., Hall, T. A.: Electron microscopy and electron probe analysis of mitochondrial cation accumulation in smooth muscle. J. Cell Biol.61, 723–742 (1974)

    Google Scholar 

  23. Vallières, J., Scarpa, A., Somlyo, A. P.: Subcellular fractions of smooth muscle. Isolation, substrate utilization and Ca++ transport by main pulmonary artery and mesenteric vein mitochondria. Arch. Biochem. Biophys.170, 659–669 (1975)

    Google Scholar 

  24. Wikström, M., Ahonen, P., Luukkainen, T.: A possible regulatory role of mitochondrial calcium uptake in uterine contractions. In: Physiology and genetics of reproduction, Vol. 4 (part B) (E. M. Coutinho and F. Fuchs, eds.), pp. 177–188 New York: Plenum Press 1974

    Google Scholar 

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Raeymaekers, L., Wuytack, F., Batra, S. et al. A comparative study of the calcium accumulation by mitochondria and microsomes isolated from the smooth muscle of the guinea-pig taenia coli. Pflugers Arch. 368, 217–223 (1977). https://doi.org/10.1007/BF00585199

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