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. 1997 Dec;73(6):3371–3384. doi: 10.1016/S0006-3495(97)78361-9

Na+ gradient-dependent Mg2+ transport in smooth muscle cells of guinea pig tenia cecum.

M Tashiro 1, M Konishi 1
PMCID: PMC1181238  PMID: 9414247

Abstract

Thin strips of guinea pig tenia cecum were loaded with the Mg2+ indicator furaptra, and the indicator fluorescence signals measured in Ca2+-free condition were converted to cytoplasmic-free Mg2+ concentration ([Mg2+]i). Lowering the extracellular Na+ concentration ([Na+]o) caused a reversible increase in [Mg2+]i, consistent with the inhibition of Na+ gradient-dependent extrusion of cellular Mg2+ (Na+-Mg2+ exchange). Curve-fitting analysis indicated that the relation between [Na+]o and the rate of rise in [Mg2+], had a Hill coefficient of approximately 3, a [Na+]o at the half-maximal rate of rise of approximately 30 mM, and a maximal rate of 0.16 +/- 0.01 microM/s (mean +/- SE, n = 6). Depolarization with 56 mM K+ shifted the curve slightly toward higher [Na+]o without significantly changing the maximal rate, suggesting that the Na+-Mg2+ exchange was inhibited by depolarization. The maximal rate would correspond to a flux of 0.15-0.4 pmol/cm2/s, if cytoplasmic Mg2+ buffering power (defined as the ratio of the changes in total Mg2+ and free Mg2+ concentrations) is assumed to be 2-5. Ouabain (1-5 microM) increased the intracellular Na+ concentration, as assessed with fluorescence of SBFI (sodium-binding benzofuran isophthalate, a Na+ indicator), and elevated [Mg2+]i. In ouabain-treated preparations, removal of extracellular Na+ rapidly increased [Mg2+]i, with an initial rate of rise roughly proportional to the degree of the Mg2+ load, and, probably, to the Na+ load caused by ouabain. The enhanced rate of rise in [Mg2+]i (up to approximately 1 microM/s) could be attributed to the Mg2+ influx as a result of the reversed Na+-Mg2+ exchange. Our results support the presence of a reversible and possibly electrogenic Na+-Mg2+ exchange in the smooth muscle cells of tenia cecum.

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Selected References

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  1. 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]
  2. Baylor S. M., Hollingworth S., Hui C. S., Quinta-Ferreira M. E. Properties of the metallochromic dyes Arsenazo III, Antipyrylazo III and Azo1 in frog skeletal muscle fibres at rest. J Physiol. 1986 Aug;377:89–141. doi: 10.1113/jphysiol.1986.sp016178. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Borin M. L., Goldman W. F., Blaustein M. P. Intracellular free Na+ in resting and activated A7r5 vascular smooth muscle cells. Am J Physiol. 1993 Jun;264(6 Pt 1):C1513–C1524. doi: 10.1152/ajpcell.1993.264.6.C1513. [DOI] [PubMed] [Google Scholar]
  4. Brading A. F. Calcium-induced increase in membrane permeability in the guinea-pig taenia coli: evidence for involvement of a sodium-calcium exchange mechanism. J Physiol. 1978 Feb;275:65–84. doi: 10.1113/jphysiol.1978.sp012178. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Brocard J. B., Rajdev S., Reynolds I. J. Glutamate-induced increases in intracellular free Mg2+ in cultured cortical neurons. Neuron. 1993 Oct;11(4):751–757. doi: 10.1016/0896-6273(93)90084-5. [DOI] [PubMed] [Google Scholar]
  6. Bülbring E., Tomita T. Effects of Ca removal on the smooth muscle of the guinea-pig taenia coli. J Physiol. 1970 Sep;210(1):217–232. doi: 10.1113/jphysiol.1970.sp009205. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Caldwell-Violich M., Requena J. Magnesium content and net fluxes in squid giant axons. J Gen Physiol. 1979 Dec;74(6):739–752. doi: 10.1085/jgp.74.6.739. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Casteels R., Kuriyama H. Membrane potential and ion content in the smooth muscle of the guinea-pig's taenia coli at different external potassium concentrations. J Physiol. 1966 May;184(1):120–130. doi: 10.1113/jphysiol.1966.sp007906. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. 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]
  10. DiPolo R., Beaugé L. An ATP-dependent Na+/Mg2+ countertransport is the only mechanism for Mg extrusion in squid axons. Biochim Biophys Acta. 1988 Dec 22;946(2):424–428. doi: 10.1016/0005-2736(88)90418-x. [DOI] [PubMed] [Google Scholar]
  11. Donoso P., Mill J. G., O'Neill S. C., Eisner D. A. Fluorescence measurements of cytoplasmic and mitochondrial sodium concentration in rat ventricular myocytes. J Physiol. 1992 Mar;448:493–509. doi: 10.1113/jphysiol.1992.sp019053. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Flatman P. W. Magnesium transport across cell membranes. J Membr Biol. 1984;80(1):1–14. doi: 10.1007/BF01868686. [DOI] [PubMed] [Google Scholar]
  13. Flatman P. W., Smith L. M. Magnesium transport in ferret red cells. J Physiol. 1990 Dec;431:11–25. doi: 10.1113/jphysiol.1990.sp018318. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Ford G. D., Driska S. P. Influence of altering cellular magnesium content on vascular smooth muscle contractility. Am J Physiol. 1986 Nov;251(5 Pt 1):C687–C695. doi: 10.1152/ajpcell.1986.251.5.C687. [DOI] [PubMed] [Google Scholar]
  15. Frenkel E. J., Graziani M., Schatzmann H. J. ATP requirement of the sodium-dependent magnesium extrusion from human red blood cells. J Physiol. 1989 Jul;414:385–397. doi: 10.1113/jphysiol.1989.sp017694. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Féray J. C., Garay R. An Na+-stimulated Mg2+-transport system in human red blood cells. Biochim Biophys Acta. 1986 Mar 27;856(1):76–84. doi: 10.1016/0005-2736(86)90012-x. [DOI] [PubMed] [Google Scholar]
  17. Féray J. C., Garay R. Demonstration of a Na+: Mg2+ exchange in human red cells by its sensitivity to tricyclic antidepressant drugs. Naunyn Schmiedebergs Arch Pharmacol. 1988 Sep;338(3):332–337. doi: 10.1007/BF00173409. [DOI] [PubMed] [Google Scholar]
  18. Gabella G. Quantitative morphological study of smooth muscle cells of the guinea-pig taenia coli. Cell Tissue Res. 1976 Jul 26;170(2):161–186. doi: 10.1007/BF00224297. [DOI] [PubMed] [Google Scholar]
  19. Gonzalez-Serratos H., Rasgado-Flores H. Extracellular magnesium-dependent sodium efflux in squid giant axons. Am J Physiol. 1990 Oct;259(4 Pt 1):C541–C548. doi: 10.1152/ajpcell.1990.259.4.C541. [DOI] [PubMed] [Google Scholar]
  20. Günther T., Vormann J. Activation of Na+/Mg2+ antiport in thymocytes by cAMP. FEBS Lett. 1992 Feb 3;297(1-2):132–134. doi: 10.1016/0014-5793(92)80343-f. [DOI] [PubMed] [Google Scholar]
  21. Günzel D., Schlue W. R. Sodium-magnesium antiport in Retzius neurones of the leech Hirudo medicinalis. J Physiol. 1996 Mar 15;491(Pt 3):595–608. doi: 10.1113/jphysiol.1996.sp021242. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Güther T., Vormann J., Förster R. Regulation of intracellular magnesium by Mg2+ efflux. Biochem Biophys Res Commun. 1984 Feb 29;119(1):124–131. doi: 10.1016/0006-291x(84)91627-9. [DOI] [PubMed] [Google Scholar]
  23. Handy R. D., Gow I. F., Ellis D., Flatman P. W. Na-dependent regulation of intracellular free magnesium concentration in isolated rat ventricular myocytes. J Mol Cell Cardiol. 1996 Aug;28(8):1641–1651. doi: 10.1006/jmcc.1996.0154. [DOI] [PubMed] [Google Scholar]
  24. Harootunian A. T., Kao J. P., Eckert B. K., Tsien R. Y. Fluorescence ratio imaging of cytosolic free Na+ in individual fibroblasts and lymphocytes. J Biol Chem. 1989 Nov 15;264(32):19458–19467. [PubMed] [Google Scholar]
  25. Hongo K., Konishi M., Kurihara S. Cytoplasmic free Mg2+ in rat ventricular myocytes studied with the fluorescent indicator furaptra. Jpn J Physiol. 1994;44(4):357–378. doi: 10.2170/jjphysiol.44.357. [DOI] [PubMed] [Google Scholar]
  26. Iwamoto T., Watano T., Shigekawa M. A novel isothiourea derivative selectively inhibits the reverse mode of Na+/Ca2+ exchange in cells expressing NCX1. J Biol Chem. 1996 Sep 13;271(37):22391–22397. doi: 10.1074/jbc.271.37.22391. [DOI] [PubMed] [Google Scholar]
  27. Kennedy R. H., Wyeth R. P., Gerner P., Liu S., Fontenot H. J., Seifen E. Tetramethylammonium is a muscarinic agonist in rat heart. Am J Physiol. 1995 Jun;268(6 Pt 1):C1414–C1417. doi: 10.1152/ajpcell.1995.268.6.C1414. [DOI] [PubMed] [Google Scholar]
  28. Kimura J. Effects of external Mg2+ on the Na-Ca exchange current in guinea pig cardiac myocytes. Ann N Y Acad Sci. 1996 Apr 15;779:515–520. doi: 10.1111/j.1749-6632.1996.tb44825.x. [DOI] [PubMed] [Google Scholar]
  29. Konishi M., Suda N., Kurihara S. Fluorescence signals from the Mg2+/Ca2+ indicator furaptra in frog skeletal muscle fibers. Biophys J. 1993 Jan;64(1):223–239. doi: 10.1016/S0006-3495(93)81359-6. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Koss K. L., Putnam R. W., Grubbs R. D. Mg2+ buffering in cultured chick ventricular myocytes: quantitation and modulation by Ca2+. Am J Physiol. 1993 May;264(5 Pt 1):C1259–C1269. doi: 10.1152/ajpcell.1993.264.5.C1259. [DOI] [PubMed] [Google Scholar]
  31. Lyu R. M., Smith L., Smith J. B. Sodium-calcium exchange in renal epithelial cells: dependence on cell sodium and competitive inhibition by magnesium. J Membr Biol. 1991 Oct;124(1):73–83. doi: 10.1007/BF01871366. [DOI] [PubMed] [Google Scholar]
  32. 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]
  33. Murphy E., Freudenrich C. C., Lieberman M. Cellular magnesium and Na/Mg exchange in heart cells. Annu Rev Physiol. 1991;53:273–287. doi: 10.1146/annurev.ph.53.030191.001421. [DOI] [PubMed] [Google Scholar]
  34. Nakayama S., Nomura H. Mechanisms of intracellular Mg2+ regulation affected by amiloride and ouabain in the guinea-pig taenia caeci. J Physiol. 1995 Oct 1;488(Pt 1):1–12. doi: 10.1113/jphysiol.1995.sp020941. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Nakayama S., Nomura H., Tomita T. Intracellular-free magnesium in the smooth muscle of guinea pig taenia caeci: a concomitant analysis for magnesium and pH upon sodium removal. J Gen Physiol. 1994 May;103(5):833–851. doi: 10.1085/jgp.103.5.833. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Nakayama S., Tomita T. Depletion of intracellular free Mg2+ in Mg2(+)- and Ca2(+)-free solution in the taenia isolated from guinea-pig caecum. J Physiol. 1990 Feb;421:363–378. doi: 10.1113/jphysiol.1990.sp017949. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Nakayama S., Tomita T. Regulation of intracellular free magnesium concentration in the taenia of guinea-pig caecum. J Physiol. 1991 Apr;435:559–572. doi: 10.1113/jphysiol.1991.sp018525. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Palatý V. Regulation of the cell magnesium in vascular smooth muscle. J Physiol. 1974 Oct;242(2):555–569. doi: 10.1113/jphysiol.1974.sp010723. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Raju B., Murphy E., Levy L. A., Hall R. D., London R. E. A fluorescent indicator for measuring cytosolic free magnesium. Am J Physiol. 1989 Mar;256(3 Pt 1):C540–C548. doi: 10.1152/ajpcell.1989.256.3.C540. [DOI] [PubMed] [Google Scholar]
  40. Romani A., Scarpa A. Regulation of cell magnesium. Arch Biochem Biophys. 1992 Oct;298(1):1–12. doi: 10.1016/0003-9861(92)90086-c. [DOI] [PubMed] [Google Scholar]
  41. Shetty S. S., Weiss G. B. Alterations in 28Mg distribution and movements in rabbit aortic smooth muscle. J Pharmacol Exp Ther. 1988 Apr;245(1):112–119. [PubMed] [Google Scholar]
  42. Stout A. K., Li-Smerin Y., Johnson J. W., Reynolds I. J. Mechanisms of glutamate-stimulated Mg2+ influx and subsequent Mg2+ efflux in rat forebrain neurones in culture. J Physiol. 1996 May 1;492(Pt 3):641–657. doi: 10.1113/jphysiol.1996.sp021334. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Tashiro M., Konishi M. Basal intracellular free Mg2+ concentration in smooth muscle cells of guinea pig tenia cecum: intracellular calibration of the fluorescent indicator furaptra. Biophys J. 1997 Dec;73(6):3358–3370. doi: 10.1016/S0006-3495(97)78360-7. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Wasserman A. J., McClellan G., Somlyo A. P. Calcium-sensitive cellular and subcellular transport of sodium, potassium, magnesium, and calcium in sodium-loaded vascular smooth muscle. Electron probe analysis. Circ Res. 1986 Jun;58(6):790–802. doi: 10.1161/01.res.58.6.790. [DOI] [PubMed] [Google Scholar]
  45. Watano T., Kimura J., Morita T., Nakanishi H. A novel antagonist, No. 7943, of the Na+/Ca2+ exchange current in guinea-pig cardiac ventricular cells. Br J Pharmacol. 1996 Oct;119(3):555–563. doi: 10.1111/j.1476-5381.1996.tb15708.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Westerblad H., Allen D. G. Myoplasmic free Mg2+ concentration during repetitive stimulation of single fibres from mouse skeletal muscle. J Physiol. 1992;453:413–434. doi: 10.1113/jphysiol.1992.sp019236. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Xu W., Willis J. S. Sodium transport through the amiloride-sensitive Na-Mg pathway of hamster red cells. J Membr Biol. 1994 Sep;141(3):277–287. doi: 10.1007/BF00235137. [DOI] [PubMed] [Google Scholar]
  48. Zhang G. H., Melvin J. E. Regulation by extracellular Na+ of cytosolic Mg2+ concentration in Mg(2+)-loaded rat sublingual acini. FEBS Lett. 1995 Aug 28;371(1):52–56. doi: 10.1016/0014-5793(95)00869-b. [DOI] [PubMed] [Google Scholar]

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