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. 1994 Apr 15;476(2):295–308. doi: 10.1113/jphysiol.1994.sp020131

Relaxation in ferret ventricular myocytes: unusual interplay among calcium transport systems.

R A Bassani 1, J W Bassani 1, D M Bers 1
PMCID: PMC1160441  PMID: 8046644

Abstract

Transport systems responsible for removing Ca2+ from the myoplasm during relaxation in isolated ferret ventricular myocytes were studied using caffeine-induced contractures. Internal calcium concentration ([Ca2+]i) was measured with the fluorescent calcium indicator indo-1, and the results were compared with our recent detailed characterizations in rabbit and rat myocytes. Relaxation and [Ca2+]i decline during a twitch in ferret myocytes were fast and similar to that in rat myocytes (i.e. half-time, t 1/2 approximately 100-160 ms). During a caffeine-induced contracture (SR Ca2+ accumulation prevented), relaxation was still relatively fast (t 1/2 = 0.57 s) and similar to relaxation in rabbit supported mainly by a strong Na(+)-Ca2+ exchange. When both the SR Ca2+ uptake and Na(+)-Ca2+ exchange are blocked (by caffeine and 0 Na+, 0 Ca2+ solution) relaxation in the ferret myocyte is remarkably fast (approximately 5-fold) compared with rabbit and rat myocytes. The decline of the Cai2+ transient was also fast under these conditions. These values were similar to those in rat under conditions where relaxation is due primarily to Na(+)-Ca2+ exchange. Additional inhibition of either the sarcolemmal Ca(2+)-ATPase or mitochondrial Ca2+ uptake caused only modest slowing of the relaxation of caffeine-induced contracture in 0 Na+, 0 Ca2+ (t 1/2 increased to approximately 3 s). In rabbit myocytes the relaxation t 1/2 is slowed to 20-30 s by these procedures. Even when the systems responsible for slow relaxation in rabbit ventricular myocytes are inhibited (i.e. sarcolemmal Ca(2+)-ATPase and mitochondrial Ca2+ uptake) along with the SR Ca(2+)-ATPase and Na(+)-Ca2+ exchange, relaxation and [Ca2+]i decline in ferret myocytes remain rapid compared with rabbit myocytes. Ca2+ taken up by mitochondria in rabbit myocytes during a caffeine contracture in 0 Na+, 0 Ca2+ solution gradually returns to the SR after caffeine removal, but this component appears to be much smaller in ferret myocytes under the same conditions. We tested for possible residual Ca2+ transport by each of the four systems which suffice to explain Ca2+ removal from the cytoplasm in rabbit (SR Ca(2+)-ATPase, Na(+)-Ca2+ exchange, sarcolemmal Ca(2+)-ATPase and mitochondrial Ca2+ uptake). We conclude that there is an additional calcium transport system at work in ferret myocytes. For this additional system, our results are most compatible with a trans-sarcolemmal Ca2+ transport, but neither a cation exchanger nor a Ca(2+)-ATPase with characteristics like that in other cardiac cells. This additional system appears able to transport Ca2+ nearly as fast as the Na(+)-Ca2+ exchange in rat ventricular myocytes.

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Selected References

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  1. Barth E., Stämmler G., Speiser B., Schaper J. Ultrastructural quantitation of mitochondria and myofilaments in cardiac muscle from 10 different animal species including man. J Mol Cell Cardiol. 1992 Jul;24(7):669–681. doi: 10.1016/0022-2828(92)93381-s. [DOI] [PubMed] [Google Scholar]
  2. 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]
  3. Bassani J. W., Bassani R. A., Bers D. M. Relaxation in rabbit and rat cardiac cells: species-dependent differences in cellular mechanisms. J Physiol. 1994 Apr 15;476(2):279–293. doi: 10.1113/jphysiol.1994.sp020130. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bassani J. W., Bassani R. A., Bers D. M. Twitch-dependent SR Ca accumulation and release in rabbit ventricular myocytes. Am J Physiol. 1993 Aug;265(2 Pt 1):C533–C540. doi: 10.1152/ajpcell.1993.265.2.C533. [DOI] [PubMed] [Google Scholar]
  5. 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]
  6. Bers D. M., Bassani R. A., Bassani J. W., Baudet S., Hryshko L. V. Paradoxical twitch potentiation after rest in cardiac muscle: increased fractional release of SR calcium. J Mol Cell Cardiol. 1993 Sep;25(9):1047–1057. doi: 10.1006/jmcc.1993.1117. [DOI] [PubMed] [Google Scholar]
  7. 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]
  8. Bers D. M., Bridge J. H., Spitzer K. W. Intracellular Ca2+ transients during rapid cooling contractures in guinea-pig ventricular myocytes. J Physiol. 1989 Oct;417:537–553. doi: 10.1113/jphysiol.1989.sp017817. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. 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]
  10. Bers D. M., Philipson K. D., Nishimoto A. Y. Sodium-calcium exchange and sidedness of isolated cardiac sarcolemmal vesicles. Biochim Biophys Acta. 1980 Sep 18;601(2):358–371. doi: 10.1016/0005-2736(80)90540-4. [DOI] [PubMed] [Google Scholar]
  11. Bers D. M., Stiffel V. M. Ratio of ryanodine to dihydropyridine receptors in cardiac and skeletal muscle and implications for E-C coupling. Am J Physiol. 1993 Jun;264(6 Pt 1):C1587–C1593. doi: 10.1152/ajpcell.1993.264.6.C1587. [DOI] [PubMed] [Google Scholar]
  12. 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]
  13. Bianchi C. P., Liu D. Calcium dependent magnesium uptake in myocardium. Life Sci. 1993;52(14):1225–1229. doi: 10.1016/0024-3205(93)90105-c. [DOI] [PubMed] [Google Scholar]
  14. Bridge J. H. Relationships between the sarcoplasmic reticulum and sarcolemmal calcium transport revealed by rapidly cooling rabbit ventricular muscle. J Gen Physiol. 1986 Oct;88(4):437–473. doi: 10.1085/jgp.88.4.437. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Bridge J. H., Spitzer K. W., Ershler P. R. Relaxation of isolated ventricular cardiomyocytes by a voltage-dependent process. Science. 1988 Aug 12;241(4867):823–825. doi: 10.1126/science.3406740. [DOI] [PubMed] [Google Scholar]
  16. Carafoli E. Calcium pump of the plasma membrane. Physiol Rev. 1991 Jan;71(1):129–153. doi: 10.1152/physrev.1991.71.1.129. [DOI] [PubMed] [Google Scholar]
  17. Cooper I. C., Fry C. H. Mechanical restitution in isolated mammalian myocardium: species differences and underlying mechanisms. J Mol Cell Cardiol. 1990 Apr;22(4):439–452. doi: 10.1016/0022-2828(90)91479-q. [DOI] [PubMed] [Google Scholar]
  18. DuBell W. H., Houser S. R. Rest decay of calcium transients and contractility in feline ventricular myocytes. Am J Physiol. 1990 Aug;259(2 Pt 2):H395–H402. doi: 10.1152/ajpheart.1990.259.2.H395. [DOI] [PubMed] [Google Scholar]
  19. Fabiato A. Time and calcium dependence of activation and inactivation of calcium-induced release of calcium from the sarcoplasmic reticulum of a skinned canine cardiac Purkinje cell. J Gen Physiol. 1985 Feb;85(2):247–289. doi: 10.1085/jgp.85.2.247. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. 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]
  21. Hove-Madsen L., Bers D. M. Passive Ca buffering and SR Ca uptake in permeabilized rabbit ventricular myocytes. Am J Physiol. 1993 Mar;264(3 Pt 1):C677–C686. doi: 10.1152/ajpcell.1993.264.3.C677. [DOI] [PubMed] [Google Scholar]
  22. Hryshko L. V., Stiffel V., Bers D. M. Rapid cooling contractures as an index of sarcoplasmic reticulum calcium content in rabbit ventricular myocytes. Am J Physiol. 1989 Nov;257(5 Pt 2):H1369–H1377. doi: 10.1152/ajpheart.1989.257.5.H1369. [DOI] [PubMed] [Google Scholar]
  23. Iino M., Kobayashi T., Endo M. Use of ryanodine for functional removal of the calcium store in smooth muscle cells of the guinea-pig. Biochem Biophys Res Commun. 1988 Apr 15;152(1):417–422. doi: 10.1016/s0006-291x(88)80730-7. [DOI] [PubMed] [Google Scholar]
  24. Inesi G., Sagara Y. Thapsigargin, a high affinity and global inhibitor of intracellular Ca2+ transport ATPases. Arch Biochem Biophys. 1992 Nov 1;298(2):313–317. doi: 10.1016/0003-9861(92)90416-t. [DOI] [PubMed] [Google Scholar]
  25. 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]
  26. Kitazawa T. Caffeine contracture in guinea-pig ventricular muscle and the effect of extracellular sodium ions. J Physiol. 1988 Aug;402:703–729. doi: 10.1113/jphysiol.1988.sp017230. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Leblanc N., Hume J. R. Sodium current-induced release of calcium from cardiac sarcoplasmic reticulum. Science. 1990 Apr 20;248(4953):372–376. doi: 10.1126/science.2158146. [DOI] [PubMed] [Google Scholar]
  28. Lehninger A. L., Carafoli E., Rossi C. S. Energy-linked ion movements in mitochondrial systems. Adv Enzymol Relat Areas Mol Biol. 1967;29:259–320. doi: 10.1002/9780470122747.ch6. [DOI] [PubMed] [Google Scholar]
  29. Lewartowski B., Wolska B. M. The effect of thapsigargin on sarcoplasmic reticulum Ca2+ content and contractions in single myocytes of guinea-pig heart. J Mol Cell Cardiol. 1993 Jan;25(1):23–29. doi: 10.1006/jmcc.1993.1004. [DOI] [PubMed] [Google Scholar]
  30. Mullins L. J. Steady-state calcium fluxes: membrane versus mitochondrial control of ionized calcium in axoplasm. Fed Proc. 1976 Dec;35(14):2583–2588. [PubMed] [Google Scholar]
  31. 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]
  32. Negretti N., O'Neill S. C., Eisner D. A. The relative contributions of different intracellular and sarcolemmal systems to relaxation in rat ventricular myocytes. Cardiovasc Res. 1993 Oct;27(10):1826–1830. doi: 10.1093/cvr/27.10.1826. [DOI] [PubMed] [Google Scholar]
  33. Näbauer M., Callewaert G., Cleemann L., Morad M. Regulation of calcium release is gated by calcium current, not gating charge, in cardiac myocytes. Science. 1989 May 19;244(4906):800–803. doi: 10.1126/science.2543067. [DOI] [PubMed] [Google Scholar]
  34. O'Neill S. C., Donoso P., Eisner D. A. The role of [Ca2+]i and [Ca2+] sensitization in the caffeine contracture of rat myocytes: measurement of [Ca2+]i and [caffeine]i. J Physiol. 1990 Jun;425:55–70. doi: 10.1113/jphysiol.1990.sp018092. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. 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]
  36. Shattock M. J., Bers D. M. Rat vs. rabbit ventricle: Ca flux and intracellular Na assessed by ion-selective microelectrodes. Am J Physiol. 1989 Apr;256(4 Pt 1):C813–C822. doi: 10.1152/ajpcell.1989.256.4.C813. [DOI] [PubMed] [Google Scholar]
  37. Smith G. L., Valdeolmillos M., Eisner D. A., Allen D. G. Effects of rapid application of caffeine on intracellular calcium concentration in ferret papillary muscles. J Gen Physiol. 1988 Sep;92(3):351–368. doi: 10.1085/jgp.92.3.351. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Thastrup O., Cullen P. J., Drøbak B. K., Hanley M. R., Dawson A. P. Thapsigargin, a tumor promoter, discharges intracellular Ca2+ stores by specific inhibition of the endoplasmic reticulum Ca2(+)-ATPase. Proc Natl Acad Sci U S A. 1990 Apr;87(7):2466–2470. doi: 10.1073/pnas.87.7.2466. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Wendt I. R., Stephenson D. G. Effects of caffeine on Ca-activated force production in skinned cardiac and skeletal muscle fibres of the rat. Pflugers Arch. 1983 Aug;398(3):210–216. doi: 10.1007/BF00657153. [DOI] [PubMed] [Google Scholar]
  40. Wolska B. M., Lewartowski B. Calcium in the in situ mitochondria of rested and stimulated myocardium. J Mol Cell Cardiol. 1991 Feb;23(2):217–226. doi: 10.1016/0022-2828(91)90108-x. [DOI] [PubMed] [Google Scholar]
  41. Wrzosek A., Schneider H., Grueninger S., Chiesi M. Effect of thapsigargin on cardiac muscle cells. Cell Calcium. 1992 May;13(5):281–292. doi: 10.1016/0143-4160(92)90063-x. [DOI] [PubMed] [Google Scholar]
  42. Yue D. T., Marban E., Wier W. G. Relationship between force and intracellular [Ca2+] in tetanized mammalian heart muscle. J Gen Physiol. 1986 Feb;87(2):223–242. doi: 10.1085/jgp.87.2.223. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Zhu Y., Nosek T. M. Inositol trisphosphate enhances Ca2+ oscillations but not Ca(2+)-induced Ca2+ release from cardiac sarcoplasmic reticulum. Pflugers Arch. 1991 Mar;418(1-2):1–6. doi: 10.1007/BF00370444. [DOI] [PubMed] [Google Scholar]

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