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. 1984 Jun 1;220(2):417–421. doi: 10.1042/bj2200417

Determination of mitochondrial calcium content in hepatocytes by a rapid cellular fractionation technique. Vasopressin stimulates mitochondrial Ca2+ uptake.

S B Shears, C J Kirk
PMCID: PMC1153642  PMID: 6743279

Abstract

Stimulation of hepatocytes with vasopressin (10 nM) in the presence of 1.25 mM extracellular Ca2+ increased glycogen phosphorylase activity 4-fold within 15s and provoked a rapid efflux of cell-associated Ca2+. Vasopressin also caused a transient increase in the Ca content of a mitochondria-rich fraction separated within seconds of hormone stimulation by a rapid fractionation technique [Shears & Kirk (1984) Biochem. J. 219, 375-382]. The Ca content of this fraction was restored to the control value within 2 min of hormone addition. These results indicate that mitochondria are not the source of the cell-associated Ca which is mobilized in the cytosol of vasopressin-stimulated hepatocytes. Rather, these organelles buffer the increase in cytosol [Ca2+] attributable to Ca mobilization from non-mitochondrial sources.

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Selected References

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  1. Althaus-Salzmann M., Carafoli E., Jakob A. Ca2+, K+ redistributions and alpha-adrenergic activation of glycogenolysis in perfused rat livers. Eur J Biochem. 1980 May;106(1):241–248. doi: 10.1111/j.1432-1033.1980.tb06015.x. [DOI] [PubMed] [Google Scholar]
  2. Balaban R. S., Blum J. J. Hormone-induced changes in NADH fluorescence and O2 consumption of rat hepatocytes. Am J Physiol. 1982 Mar;242(3):C172–C177. doi: 10.1152/ajpcell.1982.242.3.C172. [DOI] [PubMed] [Google Scholar]
  3. Barritt G. J. Evidence for two compartments of exchangeable calcium in isolated rat liver mitochondria obtained using a 45Ca exchange technique in the presence of magnesium, phosphate, and ATPase at 37 degrees C. J Membr Biol. 1981;62(1-2):53–63. doi: 10.1007/BF01870199. [DOI] [PubMed] [Google Scholar]
  4. Barritt G. J., Parker J. C., Wadsworth J. C. A kinetic analysis of the effects of adrenaline on calcium distribution in isolated rat liver parenchymal cells. J Physiol. 1981 Mar;312:29–55. doi: 10.1113/jphysiol.1981.sp013614. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Berthon B., Poggioli J., Capiod T., Claret M. Effect of the alpha-agonist noradrenaline on total and 45Ca2+ movements in mitochondria of rat liver cells. Biochem J. 1981 Oct 15;200(1):177–180. doi: 10.1042/bj2000177. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Binet A., Claret M. alpha-adrenergic stimulation of respiration in isolated rat hepatocytes. Biochem J. 1983 Mar 15;210(3):867–873. doi: 10.1042/bj2100867. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Blackmore P. F., Dehaye J. P., Exton J. H. Studies on alpha-adrenergic activation of hepatic glucose output. The role of mitochondrial calcium release in alpha-adrenergic activation of phosphorylase in perfused rat liver. J Biol Chem. 1979 Aug 10;254(15):6945–6950. [PubMed] [Google Scholar]
  8. Blackmore P. F., Hughes B. P., Charest R., Shuman E. A., 4th, Exton J. H. Time course of alpha1-adrenergic and vasopressin actions on phosphorylase activation, calcium efflux, pyridine nucleotide reduction, and respiration in hepatocytes. J Biol Chem. 1983 Sep 10;258(17):10488–10494. [PubMed] [Google Scholar]
  9. 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]
  10. Burgess G. M., McKinney J. S., Fabiato A., Leslie B. A., Putney J. W., Jr Calcium pools in saponin-permeabilized guinea pig hepatocytes. J Biol Chem. 1983 Dec 25;258(24):15336–15345. [PubMed] [Google Scholar]
  11. Creba J. A., Downes C. P., Hawkins P. T., Brewster G., Michell R. H., Kirk C. J. Rapid breakdown of phosphatidylinositol 4-phosphate and phosphatidylinositol 4,5-bisphosphate in rat hepatocytes stimulated by vasopressin and other Ca2+-mobilizing hormones. Biochem J. 1983 Jun 15;212(3):733–747. doi: 10.1042/bj2120733. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Dehaye J. P., Hughes B. P., Blackmore P. F., Exton J. H. Insulin inhibition of alpha-adrenergic actions in liver. Biochem J. 1981 Mar 15;194(3):949–956. doi: 10.1042/bj1940949. [DOI] [PMC free article] [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. Dormer R. L., Williams J. A. Secretagogue-induced changes in subcellular Ca2+ distribution in isolated pancreatic acini. Am J Physiol. 1981 Feb;240(2):G130–G140. doi: 10.1152/ajpgi.1981.240.2.G130. [DOI] [PubMed] [Google Scholar]
  15. Exton J. H. Molecular mechanisms involved in alpha-adrenergic responses. Mol Cell Endocrinol. 1981 Sep;23(3):233–264. doi: 10.1016/0303-7207(81)90123-4. [DOI] [PubMed] [Google Scholar]
  16. Hoek J. B., Nicholls D. G., Williamson J. R. Determination of the mitochondrial protonmotive force in isolated hepatocytes. J Biol Chem. 1980 Feb 25;255(4):1458–1464. [PubMed] [Google Scholar]
  17. Joseph S. K., Williamson J. R. The origin, quantitation, and kinetics of intracellular calcium mobilization by vasopressin and phenylephrine in hepatocytes. J Biol Chem. 1983 Sep 10;258(17):10425–10432. [PubMed] [Google Scholar]
  18. Keppens S., Vandenheede J. R., De Wulf H. On the role of calcium as second messenger in liver for the hormonally induced activation of glycogen phosphorylase. Biochim Biophys Acta. 1977 Feb 28;496(2):448–457. doi: 10.1016/0304-4165(77)90327-0. [DOI] [PubMed] [Google Scholar]
  19. Kirk C. J., Rodrigues L. M., Hems D. A. The influence of vasopressin and related peptides on glycogen phosphorylase activity and phosphatidylinositol metabolism in hepatocytes. Biochem J. 1979 Feb 15;178(2):493–496. doi: 10.1042/bj1780493. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. McCormack J. G., Denton R. M. Role of calcium ions in the regulation of intramitochondrial metabolism. Properties of the Ca2+-sensitive dehydrogenases within intact uncoupled mitochondria from the white and brown adipose tissue of the rat. Biochem J. 1980 Jul 15;190(1):95–105. doi: 10.1042/bj1900095. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Morgan N. G., Blackmore P. F., Exton J. H. Modulation of the alpha 1-adrenergic control of hepatocyte calcium redistribution by increases in cyclic AMP. J Biol Chem. 1983 Apr 25;258(8):5110–5116. [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. Nicholls D. G., Crompton M. Mitochondrial calcium transport. FEBS Lett. 1980 Mar 10;111(2):261–268. doi: 10.1016/0014-5793(80)80806-4. [DOI] [PubMed] [Google Scholar]
  24. Poggioli J., Berthon B., Claret M. Calcium movements in in situ mitochondria following activation of alpha-adrenergic receptors in rat liver cells. FEBS Lett. 1980 Jun 30;115(2):243–246. doi: 10.1016/0014-5793(80)81178-1. [DOI] [PubMed] [Google Scholar]
  25. 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]
  26. 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]
  27. Shears S. B., Kirk C. J. Characterization of a rapid cellular-fractionation technique for hepatocytes. Application in the measurement of mitochondrial membrane potential in situ. Biochem J. 1984 Apr 15;219(2):375–382. doi: 10.1042/bj2190375. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Shears S. B., Kirk C. J. Determination of mitochondrial calcium content in hepatocytes by a rapid cellular-fractionation technique. Alpha-adrenergic agonists do not mobilize mitochondrial Ca2+. Biochem J. 1984 Apr 15;219(2):383–389. doi: 10.1042/bj2190383. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Streb H., Irvine R. F., Berridge M. J., Schulz I. Release of Ca2+ from a nonmitochondrial intracellular store in pancreatic acinar cells by inositol-1,4,5-trisphosphate. Nature. 1983 Nov 3;306(5938):67–69. doi: 10.1038/306067a0. [DOI] [PubMed] [Google Scholar]
  30. Williamson J. R., Cooper R. H., Hoek J. B. Role of calcium in the hormonal regulation of liver metabolism. Biochim Biophys Acta. 1981 Dec 30;639(3-4):243–295. doi: 10.1016/0304-4173(81)90012-4. [DOI] [PubMed] [Google Scholar]

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