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. 1991 May;59(5):1123–1135. doi: 10.1016/S0006-3495(91)82327-X

Restoring forces in cardiac myocytes. Insight from relaxations induced by photolysis of caged ATP.

E Niggli 1, W J Lederer 1
PMCID: PMC1281347  PMID: 1868157

Abstract

Concentration jumps of intracellular ATP were produced by photolysis of P3-1-(2-nitrophenyl)ethyl (NPE)-caged ATP and were used to investigate the passive relengthening properties in unloaded cardiac myocytes. Patch-clamp pipettes in the whole-cell mode were used to voltage-clamp the myocytes and to load the cells with caged ATP while optical methods were applied to record sarcomere length or cell length simultaneously. Cell length was varied using energy deprivation contractures while intracellular Ca2+ was controlled with EGTA. At sarcomere lengths between 1.8 and 1.4 microns cellular relengthening after photolysis of caged ATP was rapid (t1/2 approximately 100 ms) and could be well described by a simple mechanical model. However, ATP jumps made at sarcomere lengths approximately 1.1 microns led to slow relengthening (t1/2 approximately seconds), comparable to the slow reextensions observed in skinned myocytes after bulk solution changes. We attribute the slow and incomplete relengthening of intact and skinned myocytes after severe rigor shortening to deformation and alteration of structural elements inside the cell. Relengthening from intermediate sarcomere lengths in intact cells is elastic and provides information about the underlying relengthening forces inside the cell. The data do not support the presence of a significant discontinuity in elastic modulus at a sarcomere length of approximately 1.6 microns expected from ultrastructural features of the sarcomeres and from observations in skinned myocytes. Our results suggest that the cell length measurements usually performed in this preparation provide an adequate description of the force produced by the unloaded cell in the steady state. The results also provide a way to estimate the error arising from viscous forces during rapid shortening.

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Selected References

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  1. Allen D. G., Morris P. G., Orchard C. H., Pirolo J. S. A nuclear magnetic resonance study of metabolism in the ferret heart during hypoxia and inhibition of glycolysis. J Physiol. 1985 Apr;361:185–204. doi: 10.1113/jphysiol.1985.sp015640. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. 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]
  3. Boyett M. R., Moore M., Jewell B. R., Montgomery R. A., Kirby M. S., Orchard C. H. An improved apparatus for the optical recording of contraction of single heart cells. Pflugers Arch. 1988 Dec;413(2):197–205. doi: 10.1007/BF00582531. [DOI] [PubMed] [Google Scholar]
  4. Bremel R. D., Weber A. Cooperation within actin filament in vertebrate skeletal muscle. Nat New Biol. 1972 Jul 26;238(82):97–101. doi: 10.1038/newbio238097a0. [DOI] [PubMed] [Google Scholar]
  5. 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]
  6. 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]
  7. Cannell M. B., Lederer W. J. A novel experimental chamber for single-cell voltage-clamp and patch-clamp applications with low electrical noise and excellent temperature and flow control. Pflugers Arch. 1986 May;406(5):536–539. doi: 10.1007/BF00583378. [DOI] [PubMed] [Google Scholar]
  8. Fabiato A., Fabiato F. Dependence of calcium release, tension generation and restoring forces on sarcomere length in skinned cardiac cells. Eur J Cardiol. 1976 May;4 (Suppl):13–27. [PubMed] [Google Scholar]
  9. Fabiato A., Fabiato F. Dependence of the contractile activation of skinned cardiac cells on the sarcomere length. Nature. 1975 Jul 3;256(5512):54–56. doi: 10.1038/256054a0. [DOI] [PubMed] [Google Scholar]
  10. Fabiato A., Fabiato F. Effects of magnesium on contractile activation of skinned cardiac cells. J Physiol. 1975 Aug;249(3):497–517. doi: 10.1113/jphysiol.1975.sp011027. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Gilbert J. C., Glantz S. A. Determinants of left ventricular filling and of the diastolic pressure-volume relation. Circ Res. 1989 May;64(5):827–852. doi: 10.1161/01.res.64.5.827. [DOI] [PubMed] [Google Scholar]
  12. Goldman Y. E., Hibberd M. G., Trentham D. R. Relaxation of rabbit psoas muscle fibres from rigor by photochemical generation of adenosine-5'-triphosphate. J Physiol. 1984 Sep;354:577–604. doi: 10.1113/jphysiol.1984.sp015394. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Gurney A. M., Lester H. A. Light-flash physiology with synthetic photosensitive compounds. Physiol Rev. 1987 Apr;67(2):583–617. doi: 10.1152/physrev.1987.67.2.583. [DOI] [PubMed] [Google Scholar]
  14. Gwathmey J. K., Copelas L., MacKinnon R., Schoen F. J., Feldman M. D., Grossman W., Morgan J. P. Abnormal intracellular calcium handling in myocardium from patients with end-stage heart failure. Circ Res. 1987 Jul;61(1):70–76. doi: 10.1161/01.res.61.1.70. [DOI] [PubMed] [Google Scholar]
  15. Hamill O. P., Marty A., Neher E., Sakmann B., Sigworth F. J. Improved patch-clamp techniques for high-resolution current recording from cells and cell-free membrane patches. Pflugers Arch. 1981 Aug;391(2):85–100. doi: 10.1007/BF00656997. [DOI] [PubMed] [Google Scholar]
  16. Hibberd M. G., Trentham D. R. Relationships between chemical and mechanical events during muscular contraction. Annu Rev Biophys Biophys Chem. 1986;15:119–161. doi: 10.1146/annurev.bb.15.060186.001003. [DOI] [PubMed] [Google Scholar]
  17. Homsher E., Millar N. C. Caged compounds and striated muscle contraction. Annu Rev Physiol. 1990;52:875–896. doi: 10.1146/annurev.ph.52.030190.004303. [DOI] [PubMed] [Google Scholar]
  18. Julian F. J., Sollins M. R. Sarcomere length-tension relations in living rat papillary muscle. Circ Res. 1975 Sep;37(3):299–308. doi: 10.1161/01.res.37.3.299. [DOI] [PubMed] [Google Scholar]
  19. Kaplan J. H., Forbush B., 3rd, Hoffman J. F. Rapid photolytic release of adenosine 5'-triphosphate from a protected analogue: utilization by the Na:K pump of human red blood cell ghosts. Biochemistry. 1978 May 16;17(10):1929–1935. doi: 10.1021/bi00603a020. [DOI] [PubMed] [Google Scholar]
  20. Krueger J. W., Forletti D., Wittenberg B. A. Uniform sarcomere shortening behavior in isolated cardiac muscle cells. J Gen Physiol. 1980 Nov;76(5):587–607. doi: 10.1085/jgp.76.5.587. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Lakatta E. G. Cardiac muscle changes in senescence. Annu Rev Physiol. 1987;49:519–531. doi: 10.1146/annurev.ph.49.030187.002511. [DOI] [PubMed] [Google Scholar]
  22. McCray J. A., Herbette L., Kihara T., Trentham D. R. A new approach to time-resolved studies of ATP-requiring biological systems; laser flash photolysis of caged ATP. Proc Natl Acad Sci U S A. 1980 Dec;77(12):7237–7241. doi: 10.1073/pnas.77.12.7237. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. McCray J. A., Trentham D. R. Properties and uses of photoreactive caged compounds. Annu Rev Biophys Biophys Chem. 1989;18:239–270. doi: 10.1146/annurev.bb.18.060189.001323. [DOI] [PubMed] [Google Scholar]
  24. Mitra R., Morad M. A uniform enzymatic method for dissociation of myocytes from hearts and stomachs of vertebrates. Am J Physiol. 1985 Nov;249(5 Pt 2):H1056–H1060. doi: 10.1152/ajpheart.1985.249.5.H1056. [DOI] [PubMed] [Google Scholar]
  25. Nichols C. G., Lederer W. J. The role of ATP in energy-deprivation contractures in unloaded rat ventricular myocytes. Can J Physiol Pharmacol. 1990 Feb;68(2):183–194. doi: 10.1139/y90-029. [DOI] [PubMed] [Google Scholar]
  26. Niggli E. A laser diffraction system with improved sensitivity for long-time measurements of sarcomere dynamics in isolated cardiac myocytes. Pflugers Arch. 1988 Apr;411(4):462–468. doi: 10.1007/BF00587728. [DOI] [PubMed] [Google Scholar]
  27. Niggli E., Lederer W. J. Real-time confocal microscopy and calcium measurements in heart muscle cells: towards the development of a fluorescence microscope with high temporal and spatial resolution. Cell Calcium. 1990 Feb-Mar;11(2-3):121–130. doi: 10.1016/0143-4160(90)90065-3. [DOI] [PubMed] [Google Scholar]
  28. Niggli E. Mechanical parameters determined in dispersed ventricular heart cells. Experientia. 1987 Dec 1;43(11-12):1150–1153. doi: 10.1007/BF01945514. [DOI] [PubMed] [Google Scholar]
  29. Periasamy A., Burns D. H., Holdren D. N., Pollack G. H., Trombitás K. A-band shortening in single fibers of frog skeletal muscle. Biophys J. 1990 Apr;57(4):815–828. doi: 10.1016/S0006-3495(90)82601-1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Roos K. P., Brady A. J. Stiffness and shortening changes in myofilament-extracted rat cardiac myocytes. Am J Physiol. 1989 Feb;256(2 Pt 2):H539–H551. doi: 10.1152/ajpheart.1989.256.2.H539. [DOI] [PubMed] [Google Scholar]
  31. Severs N. J., Slade A. M., Powell T., Twist V. W., Jones G. E. Morphometric analysis of the isolated calcium-tolerant cardiac myocyte. Organelle volumes, sarcomere length, plasma membrane surface folds, and intramembrane particle density and distribution. Cell Tissue Res. 1985;240(1):159–168. doi: 10.1007/BF00217570. [DOI] [PubMed] [Google Scholar]
  32. Siemankowski R. F., Wiseman M. O., White H. D. ADP dissociation from actomyosin subfragment 1 is sufficiently slow to limit the unloaded shortening velocity in vertebrate muscle. Proc Natl Acad Sci U S A. 1985 Feb;82(3):658–662. doi: 10.1073/pnas.82.3.658. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Tung L., Morad M. Contractile force of single heart cells compared with muscle strips of frog ventricle. Am J Physiol. 1988 Jul;255(1 Pt 2):H111–H120. doi: 10.1152/ajpheart.1988.255.1.H111. [DOI] [PubMed] [Google Scholar]
  34. Wier W. G., Cannell M. B., Berlin J. R., Marban E., Lederer W. J. Cellular and subcellular heterogeneity of [Ca2+]i in single heart cells revealed by fura-2. Science. 1987 Jan 16;235(4786):325–328. doi: 10.1126/science.3798114. [DOI] [PubMed] [Google Scholar]
  35. ter Keurs H. E., Rijnsburger W. H., van Heuningen R., Nagelsmit M. J. Tension development and sarcomere length in rat cardiac trabeculae. Evidence of length-dependent activation. Circ Res. 1980 May;46(5):703–714. doi: 10.1161/01.res.46.5.703. [DOI] [PubMed] [Google Scholar]

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