Skip to main content
NCBI home page
Environmental Health Perspectives logoLink to Environmental Health Perspectives
. 1990 Mar;84:45–56. doi: 10.1289/ehp.908445

An overview of techniques for the measurement of calcium distribution, calcium fluxes, and cytosolic free calcium in mammalian cells.

A B Borle 1
PMCID: PMC1567640  PMID: 2190818

Abstract

An array of techniques can be used to study cell calcium metabolism that comprises several calcium compartments and many types of transport systems such as ion channels, ATP-dependent pumps, and antiporters. The measurement of total cell calcium brings little information of value since 60 to 80% of total cell calcium is actually bound to the extracellular glycocalyx. Cell fractionation and differential centrifugation have been used to study intracellular Ca2+ compartmentalization, but the methods suffer from the possibility of Ca2+ loss or redistribution among cell fractions. Steady-state kinetic analyses of 45Ca uptake or desaturation curves have been used to study the distribution of Ca2+ among various kinetic pools in living cells and their rate of Ca2+ exchange, but the analyses are constrained by many limitations. Nonsteady-state tracer studies can provide information about rapid changes in calcium influx or efflux in and out of the cell. Zero-time kinetics of 45Ca uptake can detect instantaneous changes in calcium influx, while 45Ca fractional efflux ratio, can detect rapid stimulations or inhibitions of calcium efflux out of cells. Permeabilized cells have been successfully used to gauge the relative role of intracellular organelles in controlling [Ca2+]i. The measurement of the cytosolic ionized calcium ([Ca2+]i) is undoubtedly the most important and, physiologically, the most relevant method available. The choice of the appropriate calcium indicator, fluorescent, bioluminescent, metallochromic, or Ca2(+)-sensitive microelectrodes depends on the cell type and the magnitude and time constant of the event under study. Each probe has specific assets and drawbacks. The study of plasma membrane vesicles derived from baso-lateral or apical plasmalemma can also bring important information on the (Ca2(+)-Mg2+) ATPase-dependent calcium pump and on the kinetics and stoichiometry of the Na(+)-Ca2+ antiporter. The best strategy to study cell calcium metabolism is to use several different methods that focus on a specific problem from widely different angles.

Full text

PDF
45

Selected References

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

  1. Blackmore P. F., El-Refai M. F., Exton J. H. alpha-Adrenergic blockade and inhibition of A23187 mediated Ca2+ uptake by the calcium antagonist verapamil in rat liver cells. Mol Pharmacol. 1979 May;15(3):598–606. [PubMed] [Google Scholar]
  2. Blinks J. R., Wier W. G., Hess P., Prendergast F. G. Measurement of Ca2+ concentrations in living cells. Prog Biophys Mol Biol. 1982;40(1-2):1–114. doi: 10.1016/0079-6107(82)90011-6. [DOI] [PubMed] [Google Scholar]
  3. Bond M., Vadasz G., Somlyo A. V., Somlyo A. P. Subcellular calcium and magnesium mobilization in rat liver stimulated in vivo with vasopressin and glucagon. J Biol Chem. 1987 Nov 15;262(32):15630–15636. [PubMed] [Google Scholar]
  4. Borle A. B., Clark I. Effects of phosphate-induced hyperparathyroidism and parathyroidectomy on rat kidney calcium in vivo. Am J Physiol. 1981 Aug;241(2):E136–E141. doi: 10.1152/ajpendo.1981.241.2.E136. [DOI] [PubMed] [Google Scholar]
  5. Borle A. B. Control, Modulation, and regulation of cell calcium. Rev Physiol Biochem Pharmacol. 1981;90:13–153. doi: 10.1007/BFb0034078. [DOI] [PubMed] [Google Scholar]
  6. Borle A. B., Freudenrich C. C., Snowdowne K. W. A simple method for incorporating aequorin into mammalian cells. Am J Physiol. 1986 Aug;251(2 Pt 1):C323–C326. doi: 10.1152/ajpcell.1986.251.2.C323. [DOI] [PubMed] [Google Scholar]
  7. Borle A. B., Snowdowne K. W. Measurement of intracellular ionized calcium with aequorin. Methods Enzymol. 1986;124:90–116. doi: 10.1016/0076-6879(86)24011-2. [DOI] [PubMed] [Google Scholar]
  8. Borle A. B., Studer R. Effects of calcium ionophores on the transport and distribution of calcium in isolated cells and in liver and kidney slices. J Membr Biol. 1978 Jan 12;38(1-2):51–72. doi: 10.1007/BF01875162. [DOI] [PubMed] [Google Scholar]
  9. Borle A. B., Uchikawa T., Anderson J. H. Computer simulation and interpretation of 45Ca efflux profile patterns. J Membr Biol. 1982;68(1):37–46. doi: 10.1007/BF01872252. [DOI] [PubMed] [Google Scholar]
  10. 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]
  11. Reinhart P. H., Taylor W. M., Bygrave F. L. A procedure for the rapid preparation of mitochondria from rat liver. Biochem J. 1982 Jun 15;204(3):731–735. doi: 10.1042/bj2040731. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Snowdowne K. W., Borle A. B. Effects of low extracellular sodium on cytosolic ionized calcium. Na+-Ca2+ exchange as a major calcium influx pathway in kidney cells. J Biol Chem. 1985 Dec 5;260(28):14998–14507. [PubMed] [Google Scholar]
  13. Snowdowne K. W., Freudenrich C. C., Borle A. B. The effects of anoxia on cytosolic free calcium, calcium fluxes, and cellular ATP levels in cultured kidney cells. J Biol Chem. 1985 Sep 25;260(21):11619–11626. [PubMed] [Google Scholar]
  14. Studer R. K., Borle A. B. Effect of adrenalectomy on cellular calcium metabolism and on the response to adrenergic stimulation of hepatocytes isolated from male and female rats. Biochim Biophys Acta. 1984 Jul 20;804(3):377–385. doi: 10.1016/0167-4889(84)90142-3. [DOI] [PubMed] [Google Scholar]
  15. Studer R. K., Borle A. B. Effect of pH on the calcium metabolism of isolated rat kidney cells. J Membr Biol. 1979 Aug;48(4):325–341. doi: 10.1007/BF01869444. [DOI] [PubMed] [Google Scholar]
  16. Studer R. K., Borle A. B. Sex difference in cellular calcium metabolism of rat hepatocytes and in alpha-adrenergic activation of glycogen phosphorylase. Biochim Biophys Acta. 1983 Apr 5;762(2):302–314. doi: 10.1016/0167-4889(83)90085-x. [DOI] [PubMed] [Google Scholar]
  17. Studer R. K. Sexual dimorphism in adrenergic regulation of hepatic glycogenolysis. Am J Physiol. 1987 Apr;252(4 Pt 1):E467–E476. doi: 10.1152/ajpendo.1987.252.4.E467. [DOI] [PubMed] [Google Scholar]
  18. Tsien R. Y., Pozzan T., Rink T. J. Calcium homeostasis in intact lymphocytes: cytoplasmic free calcium monitored with a new, intracellularly trapped fluorescent indicator. J Cell Biol. 1982 Aug;94(2):325–334. doi: 10.1083/jcb.94.2.325. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Uchikawa T., Borle A. B. Studies of calcium-45 desaturation from kidney slices in flow-through chambers. Am J Physiol. 1978 Jan;234(1):R34–R38. doi: 10.1152/ajpregu.1978.234.1.R34. [DOI] [PubMed] [Google Scholar]
  20. Weibel E. R., Stäubli W., Gnägi H. R., Hess F. A. Correlated morphometric and biochemical studies on the liver cell. I. Morphometric model, stereologic methods, and normal morphometric data for rat liver. J Cell Biol. 1969 Jul;42(1):68–91. doi: 10.1083/jcb.42.1.68. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Environmental Health Perspectives are provided here courtesy of National Institute of Environmental Health Sciences

RESOURCES