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Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1989 Apr;86(8):2981–2984. doi: 10.1073/pnas.86.8.2981

Monitoring cytosolic free magnesium in cultured chicken heart cells by use of the fluorescent indicator Furaptra.

E Murphy 1, C C Freudenrich 1, L A Levy 1, R E London 1, M Lieberman 1
PMCID: PMC287044  PMID: 2704755

Abstract

Cytosolic free magnesium concentration [Mg2+]i and its regulation were studied in cultured embryonic chicken heart cells by use of the fluorescent indicator 2-[2-(5-carboxy)oxazole]-5-hydroxy-6-aminobenzofuran-N,N,O-triacet ic acid (Furaptra). The intracellular location of Furaptra was confirmed by its complete release from cells upon addition of saponin. The basal [Mg2+]i, which averaged 0.48 +/- 0.03 mM (n = 31), increased 3-fold on perfusion with sodium-free solution. This increase could not simply be attributed to intracellular sodium-extracellular magnesium exchange because a similar increase in [Mg2+]i occurred with magnesium-free, sodium-free perfusion. Furthermore, the increase in [Mg2+]i was largely attenuated when calcium was removed from the sodium-free perfusate. Thus, a substantial part of the increase in [Mg2+]i that occurs upon sodium-free perfusion is dependent on an increase in cytosolic free calcium (intracellular sodium-extracellular calcium exchange). The data suggest that [Mg2+]i is altered by calcium, most likely due to a competition for intracellular binding sites.

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Selected References

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  1. Altura B. M., Altura B. T. Magnesium ions and contraction of vascular smooth muscles: relationship to some vascular diseases. Fed Proc. 1981 Oct;40(12):2672–2679. [PubMed] [Google Scholar]
  2. Baker P. F., Crawford A. C. Mobility and transport of magnesium in squid giant axons. J Physiol. 1972 Dec;227(3):855–874. doi: 10.1113/jphysiol.1972.sp010062. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Blatter L. A., McGuigan J. A. Estimation of the upper limit of the free magnesium concentration measured with Mg-sensitive microelectrodes in ferret ventricular muscle: (1) use of the Nicolsky-Eisenman equation and (2) in calibrating solutions of the appropriate concentration. Magnesium. 1988;7(3):154–165. [PubMed] [Google Scholar]
  4. Blatter L. A., McGuigan J. A. Free intracellular magnesium concentration in ferret ventricular muscle measured with ion selective micro-electrodes. Q J Exp Physiol. 1986 Jul;71(3):467–473. doi: 10.1113/expphysiol.1986.sp003005. [DOI] [PubMed] [Google Scholar]
  5. Blondel B., Roijen I., Cheneval J. P. Heart cells in culture: a simple method for increasing the proportion of myoblasts. Experientia. 1971 Mar 15;27(3):356–358. doi: 10.1007/BF02138197. [DOI] [PubMed] [Google Scholar]
  6. 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]
  7. De Weer P. Axoplasmic free magnesium levels and magnesium extrusion from squid giant axons. J Gen Physiol. 1976 Aug;68(2):159–178. doi: 10.1085/jgp.68.2.159. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Eisner D. A., Orchard C. H., Allen D. G. Control of intracellular ionized calcium concentration by sarcolemmal and intracellular mechanisms. J Mol Cell Cardiol. 1984 Feb;16(2):137–146. doi: 10.1016/s0022-2828(84)80702-6. [DOI] [PubMed] [Google Scholar]
  9. Flatman P. W. Magnesium transport across cell membranes. J Membr Biol. 1984;80(1):1–14. doi: 10.1007/BF01868686. [DOI] [PubMed] [Google Scholar]
  10. Fry C. H. Measurement and control of intracellular magnesium ion concentration in guinea pig and ferret ventricular myocardium. Magnesium. 1986;5(5-6):306–316. [PubMed] [Google Scholar]
  11. GILBERT D. L. Magnesium equilibrium in muscle. J Gen Physiol. 1960 Jul;43:1103–1118. doi: 10.1085/jgp.43.6.1103. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Garfinkel L., Garfinkel D. Calculation of free-Mg2+ concentration in adenosine 5'-triphosphate containing solutions in vitro and in vivo. Biochemistry. 1984 Jul 17;23(15):3547–3552. doi: 10.1021/bi00310a025. [DOI] [PubMed] [Google Scholar]
  13. Grubbs R. D., Maguire M. E. Magnesium as a regulatory cation: criteria and evaluation. Magnesium. 1987;6(3):113–127. [PubMed] [Google Scholar]
  14. 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]
  15. Gupta R. K., Gupta P., Yushok W. D., Rose Z. B. Measurement of the dissociation constant of MgATP at physiological nucleotide levels by a combination of 31P NMR and optical absorbance spectroscopy. Biochem Biophys Res Commun. 1983 Nov 30;117(1):210–216. doi: 10.1016/0006-291x(83)91562-0. [DOI] [PubMed] [Google Scholar]
  16. Hess P., Metzger P., Weingart R. Free magnesium in sheep, ferret and frog striated muscle at rest measured with ion-selective micro-electrodes. J Physiol. 1982 Dec;333:173–188. doi: 10.1113/jphysiol.1982.sp014447. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Horie M., Irisawa H. Rectification of muscarinic K+ current by magnesium ion in guinea pig atrial cells. Am J Physiol. 1987 Jul;253(1 Pt 2):H210–H214. doi: 10.1152/ajpheart.1987.253.1.H210. [DOI] [PubMed] [Google Scholar]
  18. Meissner G., Henderson J. S. Rapid calcium release from cardiac sarcoplasmic reticulum vesicles is dependent on Ca2+ and is modulated by Mg2+, adenine nucleotide, and calmodulin. J Biol Chem. 1987 Mar 5;262(7):3065–3073. [PubMed] [Google Scholar]
  19. Mullins L. J., Brinley F. J., Jr, Spangler S. G., Abercrombie R. F. Magnesium efflux in dialyzed squid axons. J Gen Physiol. 1977 Apr;69(4):389–400. doi: 10.1085/jgp.69.4.389. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Murphy E., Wheeler D. M., LeFurgey A., Jacob R., Lobaugh L. A., Lieberman M. Coupled sodium-calcium transport in cultured chick heart cells. Am J Physiol. 1986 Mar;250(3 Pt 1):C442–C452. doi: 10.1152/ajpcell.1986.250.3.C442. [DOI] [PubMed] [Google Scholar]
  21. Page E., Polimeni P. I. Magnesium exchange in rat ventricle. J Physiol. 1972 Jul;224(1):121–139. doi: 10.1113/jphysiol.1972.sp009884. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. White R. E., Hartzell H. C. Effects of intracellular free magnesium on calcium current in isolated cardiac myocytes. Science. 1988 Feb 12;239(4841 Pt 1):778–780. doi: 10.1126/science.2448878. [DOI] [PubMed] [Google Scholar]
  23. Wu S. T., Pieper G. M., Salhany J. M., Eliot R. S. Measurement of free magnesium in perfused and ischemic arrested heart muscle. A quantitative phosphorus-31 nuclear magnetic resonance and multiequilibria analysis. Biochemistry. 1981 Dec 22;20(26):7399–7403. doi: 10.1021/bi00529a012. [DOI] [PubMed] [Google Scholar]

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