Skip to main content
NCBI home page
Biophysical Journal logoLink to Biophysical Journal
. 1996 Feb;70(2):896–916. doi: 10.1016/S0006-3495(96)79633-9

Properties of tri- and tetracarboxylate Ca2+ indicators in frog skeletal muscle fibers.

M Zhao 1, S Hollingworth 1, S M Baylor 1
PMCID: PMC1224990  PMID: 8789107

Abstract

Recently a number of lower-affinity fluorescent Ca2+ indicators have become available with principal absorbance bands at visible wavelengths. This article evaluates these indicators, as well as two shorter wavelength indicators, mag-fura-5 and mag-indo-1, for their suitability as rapid Ca2+ indicators in frog skeletal muscle fibers. With three lower-affinity tricarboxylate indicators (mag-fura-5, mag-indo-1, and magnesium orange), the change in fluorescence in response to an action potential (delta F) appeared to track the myoplasmic Ca2+ transient (delta[Ca2+]) without delay. With three lower-affinity tetracarboxylate indicators (BTC, calcium-orange-5N, and calcium-green-5N) and one tricarboxylate indicator (magnesium green), delta F responded to delta[Ca2+] with a small delay. Unfortunately, with the tetracarboxylate indicators, other problems were detected that appear to limit their usefulness as reliable Ca2+ indicators. Surprisingly, delta F from mag-fura-red, another tricarboxylate indicator, was biphasic (with 480 nm excitation), a feature that also greatly limits its usefulness. With several of the indicators, estimates were obtained for the myoplasmic value of KD, Ca (the indicator's dissociation constant for Ca2+) and found to be elevated severalfold in comparison with the value measured in a simple salt solution. These and other problems related to the quantitative use of Ca2+ indicators in the intracellular environment are evaluated and discussed.

Full text

PDF
896

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Baker A. J., Brandes R., Schreur J. H., Camacho S. A., Weiner M. W. Protein and acidosis alter calcium-binding and fluorescence spectra of the calcium indicator indo-1. Biophys J. 1994 Oct;67(4):1646–1654. doi: 10.1016/S0006-3495(94)80637-X. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bassani J. W., Bassani R. A., Bers D. M. Calibration of indo-1 and resting intracellular [Ca]i in intact rabbit cardiac myocytes. Biophys J. 1995 Apr;68(4):1453–1460. doi: 10.1016/S0006-3495(95)80318-8. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Baylor S. M., Chandler W. K., Marshall M. W. Dichroic components of Arsenazo III and dichlorophosphonazo III signals in skeletal muscle fibres. J Physiol. 1982 Oct;331:179–210. doi: 10.1113/jphysiol.1982.sp014369. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Baylor S. M., Chandler W. K., Marshall M. W. Sarcoplasmic reticulum calcium release in frog skeletal muscle fibres estimated from Arsenazo III calcium transients. J Physiol. 1983 Nov;344:625–666. doi: 10.1113/jphysiol.1983.sp014959. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Baylor S. M., Chandler W. K., Marshall M. W. Use of metallochromic dyes to measure changes in myoplasmic calcium during activity in frog skeletal muscle fibres. J Physiol. 1982 Oct;331:139–177. doi: 10.1113/jphysiol.1982.sp014368. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Baylor S. M., Hollingworth S. Absorbance signals from resting frog skeletal muscle fibers injected with the pH indicator dye, phenol red. J Gen Physiol. 1990 Sep;96(3):449–471. doi: 10.1085/jgp.96.3.449. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Baylor S. M., Hollingworth S. Fura-2 calcium transients in frog skeletal muscle fibres. J Physiol. 1988 Sep;403:151–192. doi: 10.1113/jphysiol.1988.sp017244. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. 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]
  9. Baylor S. M., Oetliker H. A large birefringence signal preceding contraction in single twitch fibres of the frog. J Physiol. 1977 Jan;264(1):141–162. doi: 10.1113/jphysiol.1977.sp011661. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Beeler T. J., Schibeci A., Martonosi A. The binding of arsenazo III to cell components. Biochim Biophys Acta. 1980 May 7;629(2):317–327. doi: 10.1016/0304-4165(80)90104-x. [DOI] [PubMed] [Google Scholar]
  11. Blinks J. R., Rüdel R., Taylor S. R. Calcium transients in isolated amphibian skeletal muscle fibres: detection with aequorin. J Physiol. 1978 Apr;277:291–323. doi: 10.1113/jphysiol.1978.sp012273. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Claflin D. R., Morgan D. L., Stephenson D. G., Julian F. J. The intracellular Ca2+ transient and tension in frog skeletal muscle fibres measured with high temporal resolution. J Physiol. 1994 Mar 1;475(2):319–325. doi: 10.1113/jphysiol.1994.sp020072. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Gillis J. M., Thomason D., Lefèvre J., Kretsinger R. H. Parvalbumins and muscle relaxation: a computer simulation study. J Muscle Res Cell Motil. 1982 Dec;3(4):377–398. doi: 10.1007/BF00712090. [DOI] [PubMed] [Google Scholar]
  14. 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]
  15. HUXLEY A. F., PEACHEY L. D. The maximum length for contraction in vertebrate straiated muscle. J Physiol. 1961 Apr;156:150–165. doi: 10.1113/jphysiol.1961.sp006665. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Harkins A. B., Kurebayashi N., Baylor S. M. Resting myoplasmic free calcium in frog skeletal muscle fibers estimated with fluo-3. Biophys J. 1993 Aug;65(2):865–881. doi: 10.1016/S0006-3495(93)81112-3. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Hirota A., Chandler W. K., Southwick P. L., Waggoner A. S. Calcium signals recorded from two new purpurate indicators inside frog cut twitch fibers. J Gen Physiol. 1989 Oct;94(4):597–631. doi: 10.1085/jgp.94.4.597. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Hollingworth S., Harkins A. B., Kurebayashi N., Konishi M., Baylor S. M. Excitation-contraction coupling in intact frog skeletal muscle fibers injected with mmolar concentrations of fura-2. Biophys J. 1992 Jul;63(1):224–234. doi: 10.1016/S0006-3495(92)81599-0. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. 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]
  20. Iatridou H., Foukaraki E., Kuhn M. A., Marcus E. M., Haugland R. P., Katerinopoulos H. E. The development of a new family of intracellular calcium probes. Cell Calcium. 1994 Feb;15(2):190–198. doi: 10.1016/0143-4160(94)90058-2. [DOI] [PubMed] [Google Scholar]
  21. Irving M., Maylie J., Sizto N. L., Chandler W. K. Simultaneous monitoring of changes in magnesium and calcium concentrations in frog cut twitch fibers containing antipyrylazo III. J Gen Physiol. 1989 Apr;93(4):585–608. doi: 10.1085/jgp.93.4.585. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Jacquemond V., Schneider M. F. Effects of low myoplasmic Mg2+ on calcium binding by parvalbumin and calcium uptake by the sarcoplasmic reticulum in frog skeletal muscle. J Gen Physiol. 1992 Jul;100(1):115–135. doi: 10.1085/jgp.100.1.115. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Kao J. P., Tsien R. Y. Ca2+ binding kinetics of fura-2 and azo-1 from temperature-jump relaxation measurements. Biophys J. 1988 Apr;53(4):635–639. doi: 10.1016/S0006-3495(88)83142-4. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Klein M. G., Simon B. J., Szucs G., Schneider M. F. Simultaneous recording of calcium transients in skeletal muscle using high- and low-affinity calcium indicators. Biophys J. 1988 Jun;53(6):971–988. doi: 10.1016/S0006-3495(88)83178-3. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Konishi M., Baylor S. M. Myoplasmic calcium transients monitored with purpurate indicator dyes injected into intact frog skeletal muscle fibers. J Gen Physiol. 1991 Feb;97(2):245–270. doi: 10.1085/jgp.97.2.245. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Konishi M., Hollingworth S., Harkins A. B., Baylor S. M. Myoplasmic calcium transients in intact frog skeletal muscle fibers monitored with the fluorescent indicator furaptra. J Gen Physiol. 1991 Feb;97(2):271–301. doi: 10.1085/jgp.97.2.271. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Konishi M., Olson A., Hollingworth S., Baylor S. M. Myoplasmic binding of fura-2 investigated by steady-state fluorescence and absorbance measurements. Biophys J. 1988 Dec;54(6):1089–1104. doi: 10.1016/S0006-3495(88)83045-5. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. 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]
  29. Kurebayashi N., Harkins A. B., Baylor S. M. Use of fura red as an intracellular calcium indicator in frog skeletal muscle fibers. Biophys J. 1993 Jun;64(6):1934–1960. doi: 10.1016/S0006-3495(93)81564-9. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Kushmerick M. J., Podolsky R. J. Ionic mobility in muscle cells. Science. 1969 Dec 5;166(3910):1297–1298. doi: 10.1126/science.166.3910.1297. [DOI] [PubMed] [Google Scholar]
  31. Lattanzio F. A., Jr, Bartschat D. K. The effect of pH on rate constants, ion selectivity and thermodynamic properties of fluorescent calcium and magnesium indicators. Biochem Biophys Res Commun. 1991 May 31;177(1):184–191. doi: 10.1016/0006-291x(91)91966-g. [DOI] [PubMed] [Google Scholar]
  32. Maylie J., Irving M., Sizto N. L., Boyarsky G., Chandler W. K. Calcium signals recorded from cut frog twitch fibers containing tetramethylmurexide. J Gen Physiol. 1987 Jan;89(1):145–176. doi: 10.1085/jgp.89.1.145. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Maylie J., Irving M., Sizto N. L., Chandler W. K. Calcium signals recorded from cut frog twitch fibers containing antipyrylazo III. J Gen Physiol. 1987 Jan;89(1):83–143. doi: 10.1085/jgp.89.1.83. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Maylie J., Irving M., Sizto N. L., Chandler W. K. Comparison of arsenazo III optical signals in intact and cut frog twitch fibers. J Gen Physiol. 1987 Jan;89(1):41–81. doi: 10.1085/jgp.89.1.41. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Minta A., Kao J. P., Tsien R. Y. Fluorescent indicators for cytosolic calcium based on rhodamine and fluorescein chromophores. J Biol Chem. 1989 May 15;264(14):8171–8178. [PubMed] [Google Scholar]
  36. Pape P. C., Jong D. S., Chandler W. K., Baylor S. M. Effect of fura-2 on action potential-stimulated calcium release in cut twitch fibers from frog muscle. J Gen Physiol. 1993 Aug;102(2):295–332. doi: 10.1085/jgp.102.2.295. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. 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]
  38. Tsien R. Y. New calcium indicators and buffers with high selectivity against magnesium and protons: design, synthesis, and properties of prototype structures. Biochemistry. 1980 May 27;19(11):2396–2404. doi: 10.1021/bi00552a018. [DOI] [PubMed] [Google Scholar]
  39. Uto A., Arai H., Ogawa Y. Reassessment of Fura-2 and the ratio method for determination of intracellular Ca2+ concentrations. Cell Calcium. 1991 Jan;12(1):29–37. doi: 10.1016/0143-4160(91)90082-p. [DOI] [PubMed] [Google Scholar]

Articles from Biophysical Journal are provided here courtesy of The Biophysical Society

RESOURCES