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. 1994 Feb 1;474(3):447–462. doi: 10.1113/jphysiol.1994.sp020036

Processes that remove calcium from the cytoplasm during excitation-contraction coupling in intact rat heart cells.

C W Balke 1, T M Egan 1, W G Wier 1
PMCID: PMC1160336  PMID: 8014906

Abstract

1. The processes that remove Ca2+ rapidly from the cytoplasm were studied in isolated rat ventricular myocytes subjected to whole-cell voltage clamp and internal perfusion with the Ca2+ indicator, indo-1. Na(+)-Ca2+ exchange was eliminated in most experiments by removing Na+ both internally and externally. 2. When the Ca(2+)-pumping ATPase of the sarcoplasmic reticulum (SR) was inhibited with cyclopiazonic acid and ryanodine interfered with the release of Ca2+ from the SR, [Ca2+]i transients rose slowly and declined extremely slowly. We concluded that transport of Ca2+ by mitochondria and the surface membrane Ca(2+)-pumping ATPase would be negligible over the time course of a single [Ca2+]i transient. 3. The influence of cytoplasmic Ca2+ ligands was characterized by internal perfusion with high concentrations of diffusible Ca2+ ligands (indo-1) or by superfusion with the membrane-permeant Ca2+ ligand, BAPTA AM. As the concentration of indo-1 in the cell increased from < 0.1 mM to at least 0.5 mM, the time constant of the decline of [Ca2+]i increased from about 0.15 s to nearly 3 s. 4. Calcium bound to endogenous Ca2+ ligands during depolarizing clamp pulses was characterized quantitatively as the difference between the total Ca2+ entering the cell via L-type Ca2+ channels and [Ca2+]i, in experiments in which SR function had been abolished. As total calcium increased during the entry of Ca2+, total calcium was found to agree reasonably well with that predicted by assuming that Ca2+ could bind to endogenous intracellular Ca2+ ligands and to indo-1. 5. The results indicate that, in the absence of Na+, the major factors determining the removal of cytoplasmic free Ca2+ are the Ca(2+)-pumping ATPase of the SR and the binding of Ca2+ to endogenous and exogenous Ca2+ ligands. 6. Several hypothetical 'Ca2+ removal functions' were fitted to the declining phase of [Ca2+]i transients. The best fit was one in which the flux of Ca2+ through the SR Ca(2+)-pumping ATPase was described by a Michaelis-Menten-type equation. The decline of the [Ca2+]i transient was thus described by a linear, first-order differential equation having terms giving the rate of Ca2+ transport by the SR Ca(2+)-pumping ATPase (Vmax and KM), the rates of complexation of Ca2+ with the various Ca2+ ligands (L), and a leak of Ca2+ into the cytoplasm from the SR (FSR,leak).(ABSTRACT TRUNCATED AT 400 WORDS)

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Selected References

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  1. Bassani J. W., Bassani R. A., Bers D. M. Ca2+ cycling between sarcoplasmic reticulum and mitochondria in rabbit cardiac myocytes. J Physiol. 1993 Jan;460:603–621. doi: 10.1113/jphysiol.1993.sp019489. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bassani R. A., Bassani J. W., Bers D. M. Mitochondrial and sarcolemmal Ca2+ transport reduce [Ca2+]i during caffeine contractures in rabbit cardiac myocytes. J Physiol. 1992;453:591–608. doi: 10.1113/jphysiol.1992.sp019246. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bers D. M., Bridge J. H., MacLeod K. T. The mechanism of ryanodine action in rabbit ventricular muscle evaluated with Ca-selective microelectrodes and rapid cooling contractures. Can J Physiol Pharmacol. 1987 Apr;65(4):610–618. doi: 10.1139/y87-103. [DOI] [PubMed] [Google Scholar]
  4. Bers D. M., Bridge J. H. Relaxation of rabbit ventricular muscle by Na-Ca exchange and sarcoplasmic reticulum calcium pump. Ryanodine and voltage sensitivity. Circ Res. 1989 Aug;65(2):334–342. doi: 10.1161/01.res.65.2.334. [DOI] [PubMed] [Google Scholar]
  5. Bers D. M., Lederer W. J., Berlin J. R. Intracellular Ca transients in rat cardiac myocytes: role of Na-Ca exchange in excitation-contraction coupling. Am J Physiol. 1990 May;258(5 Pt 1):C944–C954. doi: 10.1152/ajpcell.1990.258.5.C944. [DOI] [PubMed] [Google Scholar]
  6. Beuckelmann D. J., Wier W. G. Mechanism of release of calcium from sarcoplasmic reticulum of guinea-pig cardiac cells. J Physiol. 1988 Nov;405:233–255. doi: 10.1113/jphysiol.1988.sp017331. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Blatter L. A., Wier W. G. Intracellular diffusion, binding, and compartmentalization of the fluorescent calcium indicators indo-1 and fura-2. Biophys J. 1990 Dec;58(6):1491–1499. doi: 10.1016/S0006-3495(90)82494-2. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Cannell M. B., Berlin J. R., Lederer W. J. Effect of membrane potential changes on the calcium transient in single rat cardiac muscle cells. Science. 1987 Dec 4;238(4832):1419–1423. doi: 10.1126/science.2446391. [DOI] [PubMed] [Google Scholar]
  9. Carafoli E. The homeostasis of calcium in heart cells. J Mol Cell Cardiol. 1985 Mar;17(3):203–212. doi: 10.1016/s0022-2828(85)80003-1. [DOI] [PubMed] [Google Scholar]
  10. Crespo L. M., Grantham C. J., Cannell M. B. Kinetics, stoichiometry and role of the Na-Ca exchange mechanism in isolated cardiac myocytes. Nature. 1990 Jun 14;345(6276):618–621. doi: 10.1038/345618a0. [DOI] [PubMed] [Google Scholar]
  11. Davis B. A., Edes I., Gupta R. C., Young E. F., Kim H. W., Steenaart N. A., Szymanska G., Kranias E. G. The role of phospholamban in the regulation of calcium transport by cardiac sarcoplasmic reticulum. Mol Cell Biochem. 1990 Dec 20;99(2):83–88. doi: 10.1007/BF00230337. [DOI] [PubMed] [Google Scholar]
  12. Ehara T., Noma A., Ono K. Calcium-activated non-selective cation channel in ventricular cells isolated from adult guinea-pig hearts. J Physiol. 1988 Sep;403:117–133. doi: 10.1113/jphysiol.1988.sp017242. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Fabiato A. Calcium-induced release of calcium from the cardiac sarcoplasmic reticulum. Am J Physiol. 1983 Jul;245(1):C1–14. doi: 10.1152/ajpcell.1983.245.1.C1. [DOI] [PubMed] [Google Scholar]
  14. Frampton J. E., Orchard C. H. The effect of a calmodulin inhibitor on intracellular [Ca2+] and contraction in isolated rat ventricular myocytes. J Physiol. 1992;453:385–400. doi: 10.1113/jphysiol.1992.sp019234. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Goeger D. E., Riley R. T., Dorner J. W., Cole R. J. Cyclopiazonic acid inhibition of the Ca2+-transport ATPase in rat skeletal muscle sarcoplasmic reticulum vesicles. Biochem Pharmacol. 1988 Mar 1;37(5):978–981. doi: 10.1016/0006-2952(88)90195-5. [DOI] [PubMed] [Google Scholar]
  16. Goeger D. E., Riley R. T. Interaction of cyclopiazonic acid with rat skeletal muscle sarcoplasmic reticulum vesicles. Effect on Ca2+ binding and Ca2+ permeability. Biochem Pharmacol. 1989 Nov 15;38(22):3995–4003. doi: 10.1016/0006-2952(89)90679-5. [DOI] [PubMed] [Google Scholar]
  17. Hove-Madsen L., Bers D. M. Indo-1 binding to protein in permeabilized ventricular myocytes alters its spectral and Ca binding properties. Biophys J. 1992 Jul;63(1):89–97. doi: 10.1016/S0006-3495(92)81597-7. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Kirby M. S., Sagara Y., Gaa S., Inesi G., Lederer W. J., Rogers T. B. Thapsigargin inhibits contraction and Ca2+ transient in cardiac cells by specific inhibition of the sarcoplasmic reticulum Ca2+ pump. J Biol Chem. 1992 Jun 25;267(18):12545–12551. [PubMed] [Google Scholar]
  19. Kirchberger M. A., Tada M., Katz A. M. Adenosine 3':5'-monophosphate-dependent protein kinase-catalyzed phosphorylation reaction and its relationship to calcium transport in cardiac sarcoplasmic reticulum. J Biol Chem. 1974 Oct 10;249(19):6166–6173. [PubMed] [Google Scholar]
  20. Klein M. G., Kovacs L., Simon B. J., Schneider M. F. Decline of myoplasmic Ca2+, recovery of calcium release and sarcoplasmic Ca2+ pump properties in frog skeletal muscle. J Physiol. 1991 Sep;441:639–671. doi: 10.1113/jphysiol.1991.sp018771. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Lahouratate P., Quiniou M. J., Léoty C. Effects of cyclopiazonic acid, an inhibitor of the Ca++ ATPase of sarcoplasmic reticulum, on Ca++ transport, contraction and relaxation in cardiac muscle. Adv Exp Med Biol. 1992;311:343–345. doi: 10.1007/978-1-4615-3362-7_33. [DOI] [PubMed] [Google Scholar]
  22. Melzer W., Ríos E., Schneider M. F. The removal of myoplasmic free calcium following calcium release in frog skeletal muscle. J Physiol. 1986 Mar;372:261–292. doi: 10.1113/jphysiol.1986.sp016008. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Neher E., Augustine G. J. Calcium gradients and buffers in bovine chromaffin cells. J Physiol. 1992 May;450:273–301. doi: 10.1113/jphysiol.1992.sp019127. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Noble D., Powell T. The slowing of Ca2+ signals by Ca2+ indicators in cardiac muscle. Proc Biol Sci. 1991 Nov 22;246(1316):167–172. doi: 10.1098/rspb.1991.0140. [DOI] [PubMed] [Google Scholar]
  25. O'Neill S. C., Valdeolmillos M., Lamont C., Donoso P., Eisner D. A. The contribution of Na-Ca exchange to relaxation in mammalian cardiac muscle. Ann N Y Acad Sci. 1991;639:444–452. doi: 10.1111/j.1749-6632.1991.tb17331.x. [DOI] [PubMed] [Google Scholar]
  26. Orchard C. H., Eisner D. A., Allen D. G. Oscillations of intracellular Ca2+ in mammalian cardiac muscle. Nature. 1983 Aug 25;304(5928):735–738. doi: 10.1038/304735a0. [DOI] [PubMed] [Google Scholar]
  27. Page E. Quantitative ultrastructural analysis in cardiac membrane physiology. Am J Physiol. 1978 Nov;235(5):C147–C158. doi: 10.1152/ajpcell.1978.235.5.C147. [DOI] [PubMed] [Google Scholar]
  28. Pierce G. N., Philipson K. D., Langer G. A. Passive calcium-buffering capacity of a rabbit ventricular homogenate preparation. Am J Physiol. 1985 Sep;249(3 Pt 1):C248–C255. doi: 10.1152/ajpcell.1985.249.3.C248. [DOI] [PubMed] [Google Scholar]
  29. Seidler N. W., Jona I., Vegh M., Martonosi A. Cyclopiazonic acid is a specific inhibitor of the Ca2+-ATPase of sarcoplasmic reticulum. J Biol Chem. 1989 Oct 25;264(30):17816–17823. [PubMed] [Google Scholar]
  30. Sham J. S., Jones L. R., Morad M. Phospholamban mediates the beta-adrenergic-enhanced Ca2+ uptake in mammalian ventricular myocytes. Am J Physiol. 1991 Oct;261(4 Pt 2):H1344–H1349. doi: 10.1152/ajpheart.1991.261.4.H1344. [DOI] [PubMed] [Google Scholar]
  31. Sipido K. R., Wier W. G. Flux of Ca2+ across the sarcoplasmic reticulum of guinea-pig cardiac cells during excitation-contraction coupling. J Physiol. 1991 Apr;435:605–630. doi: 10.1113/jphysiol.1991.sp018528. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Tada M., Kadoma M., Inui M., Fujii J. Regulation of Ca2+-pump from cardiac sarcoplasmic reticulum. Methods Enzymol. 1988;157:107–154. doi: 10.1016/0076-6879(88)57073-8. [DOI] [PubMed] [Google Scholar]
  33. Takamatsu T., Wier W. G. High temporal resolution video imaging of intracellular calcium. Cell Calcium. 1990 Feb-Mar;11(2-3):111–120. doi: 10.1016/0143-4160(90)90064-2. [DOI] [PubMed] [Google Scholar]
  34. Wendt-Gallitelli M. F., Isenberg G. Total and free myoplasmic calcium during a contraction cycle: x-ray microanalysis in guinea-pig ventricular myocytes. J Physiol. 1991 Apr;435:349–372. doi: 10.1113/jphysiol.1991.sp018514. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Wier W. G. Cytoplasmic [Ca2+] in mammalian ventricle: dynamic control by cellular processes. Annu Rev Physiol. 1990;52:467–485. doi: 10.1146/annurev.ph.52.030190.002343. [DOI] [PubMed] [Google Scholar]
  36. Wier W. G., Egan T. M., López-López J. R., Balke C. W. Local control of excitation-contraction coupling in rat heart cells. J Physiol. 1994 Feb 1;474(3):463–471. doi: 10.1113/jphysiol.1994.sp020037. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Wier W. G., Kort A. A., Stern M. D., Lakatta E. G., Marban E. Cellular calcium fluctuations in mammalian heart: direct evidence from noise analysis of aequorin signals in Purkinje fibers. Proc Natl Acad Sci U S A. 1983 Dec;80(23):7367–7371. doi: 10.1073/pnas.80.23.7367. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Wier W. G., Yue D. T. Intracellular calcium transients underlying the short-term force-interval relationship in ferret ventricular myocardium. J Physiol. 1986 Jul;376:507–530. doi: 10.1113/jphysiol.1986.sp016167. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Wimsatt D. K., Hohl C. M., Brierley G. P., Altschuld R. A. Calcium accumulation and release by the sarcoplasmic reticulum of digitonin-lysed adult mammalian ventricular cardiomyocytes. J Biol Chem. 1990 Sep 5;265(25):14849–14857. [PubMed] [Google Scholar]

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