Regulation of [Na+i and [Ca2+]i during Myocardial Ischemia and Reperfusion in a Single-Cell Model

  • Hideharu Hayashi
  • Hiroshi Satoh
  • Hideki Katoh
  • Takuro Nakamura
  • Shiho Sugiyama
  • Hajime Terada
Part of the Progress in Experimental Cardiology book series (PREC, volume 1)


To study the regulation of [Na+]i and [Ca2+]i during metabolic inhibition (MI) by the perfusion of 3.3 mM amytal and 5 μM CCCP, [Na+]i and [Ca2+], were measured simultaneously using guinea pig ventricular myocytes that were dual-loaded with SBFI/M and fluo-3/AM. It was suggested that 1) [Na+]i increased during MI by both the activated Na+ influx via Na+-H+ exchange and the suppressed Na+ extrusion via the Na+-K+ pump, 2) Na+-Ca2+ exchange was inhibited during MI, causing the dissociation between [Na+]i and [Ca2+]i, 3) Na+-Ca2+ exchange could be reactivated by energy repletion, resulting in an increase of [Ca2+]i and 4) cell contracture during MI was related to rigor due to energy depletion, while cell contracture after energy repletion was likely to be related to Ca2+ overload. We also investigated the regulation of [Na+]i, [Ca2+]i, and pHi during simulated ischemia (MI with extracellular acidosis) and reperfusion. Na+-H+ exchange was active during simulated ischemia. After reperfusion, Na+-H+ exchange was activated further as pHi was recovered, resulting in an additional [Na+]i elevation.


Metabolic Inhibition Energy Depletion Cell Contracture Intracellular Acidosis Simulated Ischemia 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    Allen DG, Orchard CH. 1987. Myocardial contractile function during ischemia and hypoxia. Circ Res 60:153–168.PubMedGoogle Scholar
  2. 2.
    Lazdunski M, Frelin C, Vigne P. 1985. The sodium/hydrogen exchange system in cardiac cells: its biochemical and pharmacological properties and its role in regulating internal concentrations of sodium and internal pH. J Mol Cell Cardiol 17:1029–1042.PubMedCrossRefGoogle Scholar
  3. 3.
    January CT, Fozzard HA. 1984. The effects of membrane potential, extracellular potassium, and tetrodotoxin on the intracellular sodium ion activity of sheep cardiac muscle. Circ Res 54:652–664.PubMedGoogle Scholar
  4. 4.
    Guarnieri T. 1987. Intracellular sodium-calcium dissociation in early contractile failure in hypoxic ferret papillary muscle. J Physiol (Lond) 388:449–465.Google Scholar
  5. 5.
    Sheu SS, Fozzard HA. 1982. Transmembrane Na+ and Ca2+ electrochemical gradients in cardiac muscle and their relationship to force development. J Gen Physiol 80:325–351.PubMedCrossRefGoogle Scholar
  6. 6.
    Collins A, Somlyo AV, Hilgemann DW. 1992. The giant cardiac membrane patch method: stimulation of outward Na+-Ca2+ exchange current by Mg ATP. J Physiol (Lond) 454:27–57.Google Scholar
  7. 7.
    Doering RE, Lederer WJ. 1993. The mechanism by which cytoplasmic protons inhibit the sodium-calcium exchanger in guinea-pig heart cells. J Physiol (Lond) 466:481–499.Google Scholar
  8. 8.
    Hayashi H, Satoh H, Noda N, Terada H, Kobayashi A, Hirano M, Yamashita Y, Yamazaki N. 1994. Simultaneous measurement of intracellular Na+ and Ca2+ during K+-free perfusion in isolated myocytes. Am J Physiol 266 (Cell Physiol 35):C416–C422.PubMedGoogle Scholar
  9. 9.
    Satoh H, Hayashi H, Noda N, Terada H, Kobayashi A, Hirano M, Yamashita Y, Yamazaki N. 1994. Regulation of [Na+]i and [Ca2+]i in guinea pig myocytes: dual loading of fluorescent indicators SBFI and fluo 3. Am J Physiol 266 (Heart Circ Physiol 35):H568–H576.PubMedGoogle Scholar
  10. 10.
    Hayashi H, Miyata H. 1994. Fluorescence imaging of intracellular Ca2+. J Pharmacol Toxicol Methods 31:1–10.PubMedCrossRefGoogle Scholar
  11. 11.
    Satoh H, Hayashi H, Katoh H, Terada H, Kobayashi A. 1995. Na+/H+ and Na+/Ca2+ exchange in regulation of [Na+]i and [Ca2+]i during metabolic inhibition. Am J Physiol 268 (Heart Circ Physiol 37):H1239–H1248.PubMedGoogle Scholar
  12. 12.
    Satoh H, Hayashi H, Noda N, Terada H, Kobayashi A, Hirano M, Yamashita Y, Yamazaki N. 1991. Quantification of intracellular free sodium ions by using a new fluorescent indicator, sodium-binding benzofuran isophthalate in guinea pig myocytes. Biochem Biophys Res Commun 175:611–616.PubMedCrossRefGoogle Scholar
  13. 13.
    Miyata H, Hayashi H, Suzuki S, Noda N, Kobayashi A, Fujiwake H, Hirano M, Yamazaki N. 1989. Dual-loading of the fluorescent indicators fura-2 and 2,7-biscarboxyethyl-5(6)carboxyfluorescein (BCECF) in isolated myocytes. Biochem Biophys Res Commun 163:500–505.PubMedCrossRefGoogle Scholar
  14. 14.
    Hayashi H, Miyata H, Noda N, Kobayashi A, Hirano M, Kawai T, Yamazaki N. 1992. Intracellular Ca2+ concentration and pHi during metabolic inhibition. Am J Physiol 262 (Cell Physiol 31):C628–C634.PubMedGoogle Scholar
  15. 15.
    Li Q, Hohl CM, Altshuld RA, Stokes BT. 1989. Energy depletion-repletion and calcium transients in single cardiomyocytes. Am J Physiol 257 (Cell Physiol 26):C427–C434.PubMedGoogle Scholar
  16. 16.
    Dennis SC, Coetzee WA, Cragoe EJ Jr, Opie LH. 1990. Effects of proton buffering and of amiloride derivatives on reperfusion arrhythmias in isolated rat hearts. Circ Res 66:1156–1159.PubMedGoogle Scholar
  17. 17.
    MacLeod KT, 1989. Effects of hypoxia and metabolic inhibition on the intracellular sodium activity of mammalian ventricular muscle. J Physiol (Lond) 416:455–468.Google Scholar
  18. 18.
    Haigney MCP, Miyata H, Lakatta EG, Stern MD, Silvermann HS. 1992. Dependence of hypoxic cellular calcium loading on Na +-Ca2+ exchange. Circ Res 71:547–557.PubMedGoogle Scholar
  19. 19.
    Katoh H, Satoh H, Nakamura T, Terada H, Hayashi H. 1994. The role of Na+/H+ exchange and the Na+/K+ pump in the regulation of [Na+]i during metabolic inhibition in guinea pig myocytes. Biochem Biophys Res Commun 203:93–98.PubMedCrossRefGoogle Scholar
  20. 20.
    Russell JM, Boron WF, Brodwick MS. 1983. Intracellular pH and Na fluxes in barnacle muscle with evidence for reversal of the ionic mechanism of intracellular pH regulation. J Gen Physiol, 82:47–78.PubMedCrossRefGoogle Scholar
  21. 21.
    Miura Y, Kimura J. 1989. Sodium-calcium exchange current: dependence on internal Ca and Na and competitive binding of external Na and Ca. J Gen Physiol 93:1129–1145.PubMedCrossRefGoogle Scholar
  22. 22.
    Caroni P, Carafoli E. 1983. The regulation of the Na+-Ca2+ exchange of heart sarcolemma. Eur J Biochem 132:451–460.PubMedCrossRefGoogle Scholar
  23. 23.
    Howorth RA, Gokner AB, 1992. ATP dependence of calcium uptake by the Na-Ca exchanger of adult heart cells. Circ Res 71:210–217.Google Scholar
  24. 24.
    Rodrigo GC, Chapman RA. 1991. The calcium paradox in isolated guinea-pig ventricular myocytes: effects of membrane potential and intracellular sodium. J Physiol (Long) 434:627–645.Google Scholar
  25. 25.
    Allshire A, Piper M, Cuthbertson KSR, Cobbold PH. 1987. Cytosolic free Ca2+ in single rat heart cells during anoxia and reoxygenation. Biochem J 244:381–385.PubMedGoogle Scholar
  26. 26.
    Li Q, Atshuld RA, Stokes BT. 1988. Myocyte deenergization and intracellular free calcium dynamics. Am J Physiol 255 (Cell Physiol 24): C162–C168.PubMedGoogle Scholar
  27. 27.
    Bowers KC, Allshire AP, Cobbold PH. 1992. Bioluminescent measurement in single cardiomyocytes of sudden cytosolic ATP depletion coincident with rigor. J Mol Cell Cardiol 24:213–218.PubMedCrossRefGoogle Scholar
  28. 28.
    Anderson SE, Murphy E, Steenbergen C, London RE, Cala PM. 1990. Na-H exchange in myocardium: effects of hypoxia and acidification on Na and Ca. Am J Physiol 259 (Cell Physiol 28):C940–C948.PubMedGoogle Scholar
  29. 29.
    Tani M, Neely JR. 1989. Role of intracellular Na+ in Ca+ overload and depressed recovery of ventricular function of reperfused ischemic rat heart. Circ Res 65:1045–1056.PubMedGoogle Scholar
  30. 30.
    Cheung JY, Bonventre JV, Malis CD, Leaf A. 1986. Calcium and ischemic injury. N Engl J Med 314:1670–1676.PubMedCrossRefGoogle Scholar
  31. 31.
    Hayashi H, Miyata H, Kobayashi A, Yamazaki N. 1990. Heterogeneity in cellular response and intracellular distribution of Ca2+ concentration during metabolic inhibition. Cardiovasc Res 24:605–608.PubMedCrossRefGoogle Scholar
  32. 32.
    Nichols CG, Lederer WJ. 1990. The role of ATP in energy-deprivation contractures in unloaded rat ventricular myocytes. Can J Physiol Pharmacol 68:183–194.PubMedGoogle Scholar
  33. 33.
    Bond JM, Chacon E, Herman B, Lemasters J. 1993. Intracellular pH and Ca2+ homeostasis in the pH paradox of reperfusion injury to neonatal rat cardiac myocytes. Am J Physiol 265 (Cell Physiol 34):C129–C137.PubMedGoogle Scholar
  34. 34.
    Vaughan-Jones RD, Wu M. 1990. Extracellular H+ inactivation of Na+-H+ exchange in the sheep cardiac Purkinje fibre. J Physiol (Lond) 428:441–456Google Scholar
  35. 35.
    Ladilov V, Siegmund B, Piper HM. 1995. Protection of reoxygenated cardiomyocytes against hypercontracture by inhibition of Na+/H+ exchange. Am J Physiol (Heart Circ Physiol 37):H1531–H1539.Google Scholar

Copyright information

© Kluwer Academic Publishers 1998

Authors and Affiliations

  • Hideharu Hayashi
    • 1
  • Hiroshi Satoh
    • 1
  • Hideki Katoh
    • 1
  • Takuro Nakamura
    • 1
  • Shiho Sugiyama
    • 1
  • Hajime Terada
    • 1
  1. 1.Hamamatsu University School of MedicineJapan

Personalised recommendations