Skip to main content
NCBI home page
Biochemical Journal logoLink to Biochemical Journal
. 1983 Aug 15;214(2):405–412. doi: 10.1042/bj2140405

The effect of ionophore A23187 on calcium ion fluxes and alpha-adrenergic-agonist action in perfused rat liver.

P H Reinhart, W M Taylor, F L Bygrave
PMCID: PMC1152261  PMID: 6412701

Abstract

The effect of ionophore A23187 on cellular Ca2+ fluxes, glycogenolysis and respiration was examined in perfused liver. At low extracellular Ca2+ concentrations (less than 4 microM), A23187 induced the mobilization of intracellular Ca2+ and stimulated the rate of glycogenolysis and respiration. As the extracellular Ca2+ concentration was elevated, biphasic cellular Ca2+ fluxes were observed, with Ca2+ uptake preceding Ca2+ efflux. Under these conditions, both the glycogenolytic response and the respiratory response also became biphasic, allowing the differentiation between the effects of extracellular and intracellular Ca2+. Under all conditions examined the rate of Ca2+ efflux induced by A23187 was much slower than the rate of phenylephrine-induced Ca2+ efflux, although the net amounts of Ca2+ effluxed were similar for both agents. The effect of A23187 on phenylephrine-induced Ca2+ fluxes, glycogenolysis and respiration is dependent on the extracellular Ca2+ concentration. At concentrations of less than 50 microM-Ca2+, A23187 only partially inhibited alpha-agonist action, whereas at 1.3 mM-Ca2+ almost total inhibition was observed. The action of A23187 at the cellular level is complex, dependent on the experimental conditions used, and shows both differences from and similarities to the hepatic action of alpha-adrenergic agonists.

Full text

PDF
405

Selected References

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

  1. Akerman K. E., Nicholls D. G. Calcium transport by intact synaptosomes. Influence of ionophore A23187 on plasma-membrane potential, plasma-membrane calcium transport, mitochondrial membrane potential, respiration, cytosolic free-calcium concentration and noradrenaline release. Eur J Biochem. 1981 Mar 16;115(1):67–73. [PubMed] [Google Scholar]
  2. Assimacopoulos-Jeannet F. D., Blackmore P. F., Exton J. H. Studies on alpha-adrenergic activation of hepatic glucose output. Studies on role of calcium in alpha-adrenergic activation of phosphorylase. J Biol Chem. 1977 Apr 25;252(8):2662–2669. [PubMed] [Google Scholar]
  3. Babcock D. F., Chen J. L., Yip B. P., Lardy H. A. Evidence for mitochondrial localization of the hormone-responsive pool of Ca2+ in isolated hepatocytes. J Biol Chem. 1979 Sep 10;254(17):8117–8120. [PubMed] [Google Scholar]
  4. Babcock D. F., First N. L., Lardy H. A. Action of ionophore A23187 at the cellular level. Separation of effects at the plasma and mitochondrial membranes. J Biol Chem. 1976 Jul 10;251(13):3881–3886. [PubMed] [Google Scholar]
  5. Blackmore P. F., Brumley F. T., Marks J. L., Exton J. H. Studies on alpha-adrenergic activation of hepatic glucose output. Relationship between alpha-adrenergic stimulation of calcium efflux and activation of phosphorylase in isolated rat liver parenchymal cells. J Biol Chem. 1978 Jul 25;253(14):4851–4858. [PubMed] [Google Scholar]
  6. Blackmore P. F., Hughes B. P., Shuman E. A., Exton J. H. alpha-Adrenergic activation of phosphorylase in liver cells involves mobilization of intracellular calcium without influx of extracellular calcium. J Biol Chem. 1982 Jan 10;257(1):190–197. [PubMed] [Google Scholar]
  7. Burgess G. M., Claret M., Jenkinson D. H. Effects of catecholamines, ATP and ionophore A23187 on potassium and calcium movements in isolated hepatocytes. Nature. 1979 Jun 7;279(5713):544–546. doi: 10.1038/279544a0. [DOI] [PubMed] [Google Scholar]
  8. Chandler D. E., Williams J. A. Intracellular divalent cation release in pancreatic acinar cells during stimulus-secretion coupling. II. Subcellular localization of the fluorescent probe chlorotetracycline. J Cell Biol. 1978 Feb;76(2):386–399. doi: 10.1083/jcb.76.2.386. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Chen J. L., Babcock D. F., Lardy H. A. Norepinephrine, vasopressin, glucagon, and A23187 induce efflux of calcium from an exchangeable pool in isolated rat hepatocytes. Proc Natl Acad Sci U S A. 1978 May;75(5):2234–2238. doi: 10.1073/pnas.75.5.2234. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Coll K. E., Joseph S. K., Corkey B. E., Williamson J. R. Determination of the matrix free Ca2+ concentration and kinetics of Ca2+ efflux in liver and heart mitochondria. J Biol Chem. 1982 Aug 10;257(15):8696–8704. [PubMed] [Google Scholar]
  11. Deber C. M., Pfieffer D. R. Ionophore A23187. Solution conformations of the calcium complex and free acid deduced from proton and carbon-13 nuclear magnetic resonance studies. Biochemistry. 1976 Jan 13;15(1):132–141. doi: 10.1021/bi00646a020. [DOI] [PubMed] [Google Scholar]
  12. Dehaye J. P., Blackmore P. F., Venter J. C., Exton J. H. Studies on the alpha-adrenergic activation of hepatic glucose output. alpha-Adrenergic activation of phosphorylase by immobilized epinephrine. J Biol Chem. 1980 May 10;255(9):3905–3910. [PubMed] [Google Scholar]
  13. Denton R. M., McCormack J. G., Edgell N. J. Role of calcium ions in the regulation of intramitochondrial metabolism. Effects of Na+, Mg2+ and ruthenium red on the Ca2+-stimulated oxidation of oxoglutarate and on pyruvate dehydrogenase activity in intact rat heart mitochondria. Biochem J. 1980 Jul 15;190(1):107–117. doi: 10.1042/bj1900107. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Foden S., Randle P. J. Calcium metabolism in rat hepatocytes. Biochem J. 1978 Mar 15;170(3):615–625. doi: 10.1042/bj1700615. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Friedmann N., Divakaran P., Kirkland J., Kimura S., Wood J. Effects of the calcium ionophore A23187 on liver metabolism. J Pharmacol Exp Ther. 1979 Oct;211(1):127–132. [PubMed] [Google Scholar]
  16. Hasper F., Müller G., Schweizer J. Histological, proliferative, and biochemical alterations in dorsal epidermis of the Syrian golden hamster induced by 12-O-tetradecanoylphorbol-13-acetate and the calcium ionophore A 23187. Cancer Res. 1982 May;42(5):2034–2039. [PubMed] [Google Scholar]
  17. Haye B., Marcy G., Jacquemin C. Relationship between the "phospholipid effect" and calcium in the thyroid. I. - Effects of calcium ions, E.G.T.A., ionophore A 23187, verapamil and chlorpromazine on resting and stimulate thyroid slices. Biochimie. 1979;61(8):905–912. doi: 10.1016/s0300-9084(79)80240-0. [DOI] [PubMed] [Google Scholar]
  18. Heaton G. M., Nicholls D. G. The calcium conductance of the inner membrane of rat liver mitochondria and the determination of the calcium electrochemical gradient. Biochem J. 1976 Jun 15;156(3):635–646. doi: 10.1042/bj1560635. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Khoo J. C., Steinberg D. Stimulation of rat liver phosphorylase kinase by micromolar concentrations of Ca2+. FEBS Lett. 1975 Sep 1;57(1):68–72. doi: 10.1016/0014-5793(75)80154-2. [DOI] [PubMed] [Google Scholar]
  20. Kleineke J., Stratman F. W. Calcium transport in isolated rat hepatocytes. FEBS Lett. 1974 Jul 1;43(1):75–80. doi: 10.1016/0014-5793(74)81109-9. [DOI] [PubMed] [Google Scholar]
  21. McCormack J. G., Edgell N. J., Denton R. M. Studies on the interactions of Ca2+ and pyruvate in the regulation of rat heart pyruvate dehydrogenase activity. Effects of starvation and diabetes. Biochem J. 1982 Feb 15;202(2):419–427. doi: 10.1042/bj2020419. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Murphy E., Coll K., Rich T. L., Williamson J. R. Hormonal effects on calcium homeostasis in isolated hepatocytes. J Biol Chem. 1980 Jul 25;255(14):6600–6608. [PubMed] [Google Scholar]
  23. Reed P. W., Lardy H. A. A23187: a divalent cation ionophore. J Biol Chem. 1972 Nov 10;247(21):6970–6977. [PubMed] [Google Scholar]
  24. Reinhart P. H., Taylor W. M., Bygrave F. L. Calcium ion fluxes induced by the action of alpha-adrenergic agonists in perfused rat liver. Biochem J. 1982 Dec 15;208(3):619–630. doi: 10.1042/bj2080619. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Reinhart P. H., Taylor W. M., Bygrave F. L. Studies on alpha-adrenergic-induced respiration and glycogenolysis in perfused rat liver. J Biol Chem. 1982 Feb 25;257(4):1906–1912. [PubMed] [Google Scholar]
  26. Sakai K., Matsumura S., Okimura Y., Yamamura H., Nishizuka Y. Liver glycogen phosphorylase kinase. Partial purification and characterization. J Biol Chem. 1979 Jul 25;254(14):6631–6637. [PubMed] [Google Scholar]
  27. Schuster S. M., Olson M. S. Studies of the energy-dependent uptake of divalent metal ions by beef heart mitochondria. J Biol Chem. 1974 Nov 25;249(22):7151–7158. [PubMed] [Google Scholar]
  28. Selinger Z., Eimerl S., Schramm M. A calcium ionophore simulating the action of epinephrine on the alpha-adrenergic receptor. Proc Natl Acad Sci U S A. 1974 Jan;71(1):128–131. doi: 10.1073/pnas.71.1.128. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Shimazu T., Amakawa A. Regulation of glycogen metabolism in liver by the autonomic nervous system. VI. Possible mechanism of phosphorylase activation by the splanchnic nerve. Biochim Biophys Acta. 1975 Apr 7;385(2):242–256. doi: 10.1016/0304-4165(75)90352-9. [DOI] [PubMed] [Google Scholar]
  30. Sordahl L. A. Effects of magnesium, Ruthenium red and the antibiotic ionophore A-23187 on initial rates of calcium uptake and release by heart mitochondria. Arch Biochem Biophys. 1975 Mar;167(1):104–115. doi: 10.1016/0003-9861(75)90446-4. [DOI] [PubMed] [Google Scholar]
  31. Sugden P. H. The effects of calcium ions, ionophore A23187 and inhibition of energy metabolism on protein degradation in the rat diaphragm and epitrochlearis muscles in vitro. Biochem J. 1980 Sep 15;190(3):593–603. doi: 10.1042/bj1900593. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Whiting J. A., Barritt G. J. On the mechanism by which hormones induce the release of Ca2+ from mitochondria in the liver cell. Biochem J. 1982 Jul 15;206(1):121–129. doi: 10.1042/bj2060121. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Wong D. T., Wilkinson J. R., Hamill R. L., Horng J. S. Effects of antibiotic ionophore, A23187, on oxidative phosphorylation and calcium transport of liver mitochondria. Arch Biochem Biophys. 1973 Jun;156(2):578–585. doi: 10.1016/0003-9861(73)90308-1. [DOI] [PubMed] [Google Scholar]

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

RESOURCES