Skip to main content
Biochemical Journal logoLink to Biochemical Journal
. 1987 Dec 1;248(2):609–613. doi: 10.1042/bj2480609

Measurement of the matrix free Ca2+ concentration in heart mitochondria by entrapped fura-2 and quin2.

G L Lukács 1, A Kapus 1
PMCID: PMC1148585  PMID: 3435469

Abstract

A method was developed to monitor continuously the matrix free Ca2+ concentration ([Ca2+]m) of heart mitochondria by use of the fluorescent Ca2+ indicators, fura-2 and quin2. The acetoxymethyl esters of fura-2 and quin2 were accumulated in and hydrolysed by isolated mitochondria. An increase of the mitochondrial Ca content from 0.3 nmol/mg of protein to 6 nmol/mg corresponded to a rise of [Ca2+]m from 30 to 1000 nM. The results indicate that physiological fluctuations of the mitochondrial Ca content elicit changes of [Ca2+]m in that range which regulates the matrix dehydrogenases.

Full text

PDF
609

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Arslan P., Di Virgilio F., Beltrame M., Tsien R. Y., Pozzan T. Cytosolic Ca2+ homeostasis in Ehrlich and Yoshida carcinomas. A new, membrane-permeant chelator of heavy metals reveals that these ascites tumor cell lines have normal cytosolic free Ca2+. J Biol Chem. 1985 Mar 10;260(5):2719–2727. [PubMed] [Google Scholar]
  2. Ashley R. H. Buffer capacity of intracellular Ca2+ indicators. Biochem J. 1986 Nov 15;240(1):310–311. doi: 10.1042/bj2400310. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Assimacopoulos-Jeannet F., McCormack J. G., Jeanrenaud B. Vasopressin and/or glucagon rapidly increases mitochondrial calcium and oxidative enzyme activities in the perfused rat liver. J Biol Chem. 1986 Jul 5;261(19):8799–8804. [PubMed] [Google Scholar]
  4. Coll K. E., Joseph S. K., Corkey B. E., Williamson J. R. Determination of the matrix free Ca2+ concentration and kinetics of Ca2+ efflux in liver and heart mitochondria. J Biol Chem. 1982 Aug 10;257(15):8696–8704. [PubMed] [Google Scholar]
  5. Corkey B. E., Duszynski J., Rich T. L., Matschinsky B., Williamson J. R. Regulation of free and bound magnesium in rat hepatocytes and isolated mitochondria. J Biol Chem. 1986 Feb 25;261(6):2567–2574. [PubMed] [Google Scholar]
  6. Crompton M., Kessar P., Al-Nasser I. The alpha-adrenergic-mediated activation of the cardiac mitochondrial Ca2+ uniporter and its role in the control of intramitochondrial Ca2+ in vivo. Biochem J. 1983 Nov 15;216(2):333–342. doi: 10.1042/bj2160333. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Denton R. M., McCormack J. G. Ca2+ transport by mammalian mitochondria and its role in hormone action. Am J Physiol. 1985 Dec;249(6 Pt 1):E543–E554. doi: 10.1152/ajpendo.1985.249.6.E543. [DOI] [PubMed] [Google Scholar]
  8. Denton R. M., McCormack J. G. On the role of the calcium transport cycle in heart and other mammalian mitochondria. FEBS Lett. 1980 Sep 22;119(1):1–8. doi: 10.1016/0014-5793(80)80986-0. [DOI] [PubMed] [Google Scholar]
  9. Grynkiewicz G., Poenie M., Tsien R. Y. A new generation of Ca2+ indicators with greatly improved fluorescence properties. J Biol Chem. 1985 Mar 25;260(6):3440–3450. [PubMed] [Google Scholar]
  10. Hansford R. G., Castro F. Intramitochondrial and extramitochondrial free calcium ion concentrations of suspensions of heart mitochondria with very low, plausibly physiological, contents of total calcium. J Bioenerg Biomembr. 1982 Dec;14(5-6):361–376. doi: 10.1007/BF00743064. [DOI] [PubMed] [Google Scholar]
  11. Hansford R. G. Relation between mitochondrial calcium transport and control of energy metabolism. Rev Physiol Biochem Pharmacol. 1985;102:1–72. doi: 10.1007/BFb0034084. [DOI] [PubMed] [Google Scholar]
  12. Hesketh T. R., Smith G. A., Moore J. P., Taylor M. V., Metcalfe J. C. Free cytoplasmic calcium concentration and the mitogenic stimulation of lymphocytes. J Biol Chem. 1983 Apr 25;258(8):4876–4882. [PubMed] [Google Scholar]
  13. Ligeti E., Lukács G. L. Phosphate transport, membrane potential, and movements of calcium in rat liver mitochondria. J Bioenerg Biomembr. 1984 Apr;16(2):101–113. doi: 10.1007/BF00743043. [DOI] [PubMed] [Google Scholar]
  14. Luft J. H. Ruthenium red and violet. I. Chemistry, purification, methods of use for electron microscopy and mechanism of action. Anat Rec. 1971 Nov;171(3):347–368. doi: 10.1002/ar.1091710302. [DOI] [PubMed] [Google Scholar]
  15. Lukács G. L., Fonyó A. The Ba2+ sensitivity of the Na+-induced Ca2+ efflux in heart mitochondria: the site of inhibitory action. Biochim Biophys Acta. 1986 Jun 13;858(1):125–134. doi: 10.1016/0005-2736(86)90298-1. [DOI] [PubMed] [Google Scholar]
  16. Moore C. L. Specific inhibition of mitochondrial Ca++ transport by ruthenium red. Biochem Biophys Res Commun. 1971 Jan 22;42(2):298–305. doi: 10.1016/0006-291x(71)90102-1. [DOI] [PubMed] [Google Scholar]
  17. Pollock W. K., Rink T. J., Irvine R. F. Liberation of [3H]arachidonic acid and changes in cytosolic free calcium in fura-2-loaded human platelets stimulated by ionomycin and collagen. Biochem J. 1986 May 1;235(3):869–877. doi: 10.1042/bj2350869. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Puskin J. S., Gunter T. E., Gunter K. K., Russell P. R. Evidence for more than one Ca2+ transport mechanism in mitochondria. Biochemistry. 1976 Aug 24;15(17):3834–3842. doi: 10.1021/bi00662a029. [DOI] [PubMed] [Google Scholar]
  19. Reinhart P. H., van de Pol E., Taylor W. M., Bygrave F. L. An assessment of the calcium content of rat liver mitochondria in vivo. Biochem J. 1984 Mar 1;218(2):415–420. doi: 10.1042/bj2180415. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Schackmann R. W., Chock P. B. Alteration of intracellular [Ca2+] in sea urchin sperm by the egg peptide speract. Evidence that increased intracellular Ca2+ is coupled to Na+ entry and increased intracellular pH. J Biol Chem. 1986 Jul 5;261(19):8719–8728. [PubMed] [Google Scholar]
  21. Somlyo A. P., Bond M., Somlyo A. V. Calcium content of mitochondria and endoplasmic reticulum in liver frozen rapidly in vivo. Nature. 1985 Apr 18;314(6012):622–625. doi: 10.1038/314622a0. [DOI] [PubMed] [Google Scholar]
  22. Spät A., Lukács G. L., Eberhardt I., Kiesel L., Runnebaum B. Binding of inositol phosphates and induction of Ca2+ release from pituitary microsomal fractions. Biochem J. 1987 Jun 1;244(2):493–496. doi: 10.1042/bj2440493. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Tsien R. Y. A non-disruptive technique for loading calcium buffers and indicators into cells. Nature. 1981 Apr 9;290(5806):527–528. doi: 10.1038/290527a0. [DOI] [PubMed] [Google Scholar]
  24. Tsien R. Y., Pozzan T., Rink T. J. Calcium homeostasis in intact lymphocytes: cytoplasmic free calcium monitored with a new, intracellularly trapped fluorescent indicator. J Cell Biol. 1982 Aug;94(2):325–334. doi: 10.1083/jcb.94.2.325. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Wendt-Gallitelli M. F., Jacob R. Rhythm-dependent role of different calcium stores in cardiac muscle: X-ray microanalysis. J Mol Cell Cardiol. 1982 Aug;14(8):487–492. doi: 10.1016/0022-2828(82)90157-2. [DOI] [PubMed] [Google Scholar]
  26. Williams D. A., Fogarty K. E., Tsien R. Y., Fay F. S. Calcium gradients in single smooth muscle cells revealed by the digital imaging microscope using Fura-2. Nature. 1985 Dec 12;318(6046):558–561. doi: 10.1038/318558a0. [DOI] [PubMed] [Google Scholar]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society

RESOURCES