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. 1986 Jul 15;237(2):447–454. doi: 10.1042/bj2370447

Increase of adenylate cyclase catalytic-unit activity by dexamethasone in rat osteoblast-like cells.

R Rizzoli, V von Tscharner, H Fleisch
PMCID: PMC1147005  PMID: 3099755

Abstract

Glucocorticoids are known to increase the cyclic AMP response to parathyroid hormone (PTH) in cultured bone organs or bone cells. Using the osteoblast-like cell line ROS 17/2.8, which possesses receptors for both PTH and glucocorticoids, we investigated which component of the complex hormone receptor-guanine nucleotide regulatory unit--adenylate cyclase was affected by dexamethasone treatment. In response to PTH, isoproterenol or forskolin, a compound that is supposed to act directly on the catalytic unit, cyclic AMP production by intact cells and adenylate cyclase activity in purified plasma membrane were markedly increased by dexamethasone. Whereas NaF, guanosine 5'-[beta gamma-imido]triphosphate and Mn/ stimulated adenylate cyclase activity were similarly enhanced in membranes isolated from glucocorticoid-treated cells, the activity of the stimulatory guanine nucleotide regulatory unit, as assessed by reconstitution into membranes from the CYC- clone, which is genetically devoid of this component, was not altered. Thus in osteoblast-like cells dexamethasone appears to increase cyclic AMP synthesis by influencing the catalytic unit. Moreover, since it has been reported that glucocorticoids may produce changes in cell calcium metabolism, we evaluated cytoplasmic free Ca2+ concentration ([Ca2+]i) and intracellular Ca2+ stores mobilizable by the bivalent-cationophore ionomycin, by using the intracellular fluorescent indicator Quin-2. The results indicated that dexamethasone treatment did not influence [Ca2+]i but markedly decreased ionomycin-releasable Ca2+ stores.

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Selected References

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

  1. BURTON K. A study of the conditions and mechanism of the diphenylamine reaction for the colorimetric estimation of deoxyribonucleic acid. Biochem J. 1956 Feb;62(2):315–323. doi: 10.1042/bj0620315. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Berthon B., Binet A., Mauger J. P., Claret M. Cytosolic free Ca2+ in isolated rat hepatocytes as measured by quin2. Effects of noradrenaline and vasopressin. FEBS Lett. 1984 Feb 13;167(1):19–24. doi: 10.1016/0014-5793(84)80824-8. [DOI] [PubMed] [Google Scholar]
  3. Bourne H. R., Coffino P., Tomkins G. M. Selection of a variant lymphoma cell deficient in adenylate cyclase. Science. 1975 Feb 28;187(4178):750–752. doi: 10.1126/science.163487. [DOI] [PubMed] [Google Scholar]
  4. Brown B. L., Albano J. D., Ekins R. P., Sgherzi A. M. A simple and sensitive saturation assay method for the measurement of adenosine 3':5'-cyclic monophosphate. Biochem J. 1971 Feb;121(3):561–562. doi: 10.1042/bj1210561. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Catherwood B. D. 1,25-Dihydroxycholecalciferol and glucocorticosteroid regulation of adenylate cyclase in an osteoblast-like cell line. J Biol Chem. 1985 Jan 25;260(2):736–743. [PubMed] [Google Scholar]
  6. Chen T. L., Feldman D. Glucocorticoid potentiation of the adenosin 3',5'-monophosphate response to parathyroid hormone in cultured rat bone cells. Endocrinology. 1978 Feb;102(2):589–596. doi: 10.1210/endo-102-2-589. [DOI] [PubMed] [Google Scholar]
  7. Chen T. L., Feldman D. Glucocorticoid receptors and actions in subpopulations of cultured rat bone cells. Mechanism of dexamethasone potentiation of parathyroid hormone-stimulated cyclic AMP production. J Clin Invest. 1979 Apr;63(4):750–758. doi: 10.1172/JCI109359. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Chen T. L., Feldman D. Modulation of PTH-stimulated cyclic AMP in cultured rodent bone cells: the effects of 1,25(OH)2 vitamin D3 and its interaction with glucocorticoids. Calcif Tissue Int. 1984 Sep;36(5):580–585. doi: 10.1007/BF02405370. [DOI] [PubMed] [Google Scholar]
  9. Clark R. B., Goka T. J., Green D. A., Barber R., Butcher R. W. Differences in the forskolin activation of adenylate cyclases in wild-type and variant lymphoma cells. Mol Pharmacol. 1982 Nov;22(3):609–613. [PubMed] [Google Scholar]
  10. Davies A. O., Lefkowitz R. J. Regulation of beta-adrenergic receptors by steroid hormones. Annu Rev Physiol. 1984;46:119–130. doi: 10.1146/annurev.ph.46.030184.001003. [DOI] [PubMed] [Google Scholar]
  11. Dietrich J. W., Canalis E. M., Maina D. M., Raisz L. G. Hormonal control of bone collagen synthesis in vitro: effects of parathyroid hormone and calcitonin. Endocrinology. 1976 Apr;98(4):943–949. doi: 10.1210/endo-98-4-943. [DOI] [PubMed] [Google Scholar]
  12. Downs R. W., Jr, Aurbach G. D. The effects of forskolin on adenylate cyclase in S49 wild type and cyc- cells. J Cyclic Nucleotide Res. 1982;8(4):235–242. [PubMed] [Google Scholar]
  13. Downs R. W., Jr, Levine M. A., Drezner M. K., Burch W. M., Jr, Spiegel A. M. Deficient adenylate cyclase regulatory protein in renal membranes from a patient with pseudohypoparathyroidism. J Clin Invest. 1983 Feb;71(2):231–235. doi: 10.1172/JCI110763. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Egan J. J., Majeska R. J., Rodan G. A. Osteosarcoma cytosol factor promotes parathyroid hormone stimulation of adenylate cyclase independent of GTP. Biochim Biophys Acta. 1980 Nov 3;632(4):483–496. doi: 10.1016/0304-4165(80)90325-6. [DOI] [PubMed] [Google Scholar]
  15. Foster S. J., Perkins J. P. Glucocorticoids increase the responsiveness of cells in culture to prostaglandin E1. Proc Natl Acad Sci U S A. 1977 Nov;74(11):4816–4820. doi: 10.1073/pnas.74.11.4816. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Gilman A. G. G proteins and dual control of adenylate cyclase. Cell. 1984 Mar;36(3):577–579. doi: 10.1016/0092-8674(84)90336-2. [DOI] [PubMed] [Google Scholar]
  17. Grinstein S., Elder B., Clarke C. A., Buchwald M. Is cytoplasmic Ca2+ in lymphocytes elevated in cystic fibrosis? Biochim Biophys Acta. 1984 Jan 11;769(1):270–274. doi: 10.1016/0005-2736(84)90032-4. [DOI] [PubMed] [Google Scholar]
  18. Hahn T. J., Halstead L. R. Cortisol enhancement of PTH-stimulated cyclic AMP accumulation in cultured fetal rat long bone rudiments. Calcif Tissue Int. 1979 Nov 26;29(2):173–175. doi: 10.1007/BF02408074. [DOI] [PubMed] [Google Scholar]
  19. Haussler M. R., Manolagas S. C., Deftos L. J. Glucocorticoid receptor in clonal osteosarcoma cell lines: a novel system for investigating bone active hormones. Biochem Biophys Res Commun. 1980 May 14;94(1):373–380. doi: 10.1016/s0006-291x(80)80231-2. [DOI] [PubMed] [Google Scholar]
  20. Hällgren R., Svenson K., Johansson E., Lindh U. Abnormal calcium and magnesium stores in erythrocytes and granulocytes from patients with inflammatory connective tissue diseases. Relationship to inflammatory activity and effect of corticosteroid therapy. Arthritis Rheum. 1985 Feb;28(2):169–173. doi: 10.1002/art.1780280211. [DOI] [PubMed] [Google Scholar]
  21. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  22. MacNeil S., Crawford A., Amirrasooli H., Johnson S., Pollock A., Ollis C., Tomlinson S. Stimulation of hormone-responsive adenylate cyclase activity by a factor present in the cell cytosol. Biochem J. 1980 May 15;188(2):393–400. doi: 10.1042/bj1880393. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Majeska R. J., Rodan S. B., Rodan G. A. Parathyroid hormone-responsive clonal cell lines from rat osteosarcoma. Endocrinology. 1980 Nov;107(5):1494–1503. doi: 10.1210/endo-107-5-1494. [DOI] [PubMed] [Google Scholar]
  24. Manganiello V., Vaughan M. An effect of dexamethasone on adenosine 3',5'-monophosphate content and adenosine 3',5'-monophosphate phosphodiesterase activity of cultured hepatoma cells. J Clin Invest. 1972 Oct;51(10):2763–2767. doi: 10.1172/JCI107096. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Ng B., Hekkelman J. W., Heersche J. N. The effect of cortisol on the adenosine 3',5'-monophosphate response to parathyroid hormone of bone in vitro. Endocrinology. 1979 Apr;104(4):1130–1135. doi: 10.1210/endo-104-4-1130. [DOI] [PubMed] [Google Scholar]
  26. Peck W. A., Kohler G., Barr S. Calcium-mediated enhancement of the cyclic AMP response in cultured bone cells. Calcif Tissue Int. 1981;33(4):409–416. doi: 10.1007/BF02409464. [DOI] [PubMed] [Google Scholar]
  27. Rasmussen H., Gustin M. C. Some aspects of the hormonal control of cellular calcium metabolism. Ann N Y Acad Sci. 1978 Apr 28;307:391–401. doi: 10.1111/j.1749-6632.1978.tb41964.x. [DOI] [PubMed] [Google Scholar]
  28. Rizzoli R. E., Somerman M., Murray T. M., Aurbach G. D. Binding of radioiodinated parathyroid hormone to cloned bone cells. Endocrinology. 1983 Nov;113(5):1832–1838. doi: 10.1210/endo-113-5-1832. [DOI] [PubMed] [Google Scholar]
  29. Rodan S. B., Fischer M. K., Egan J. J., Epstein P. M., Rodan G. A. The effect of dexamethasone on parathyroid hormone stimulation of adenylate cyclase in ROS 17/2.8 cells. Endocrinology. 1984 Sep;115(3):951–958. doi: 10.1210/endo-115-3-951. [DOI] [PubMed] [Google Scholar]
  30. Rottenberg H. The measurement of membrane potential and deltapH in cells, organelles, and vesicles. Methods Enzymol. 1979;55:547–569. doi: 10.1016/0076-6879(79)55066-6. [DOI] [PubMed] [Google Scholar]
  31. Sahyoun N., LeVine H., 3rd, Stenbuck P., Cuatrecasas P. Cytosolic activator of adenylate cyclase: Reconstitution, characterization, and mechanism of action. Proc Natl Acad Sci U S A. 1983 Jun;80(12):3646–3650. doi: 10.1073/pnas.80.12.3646. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Salomon Y., Londos C., Rodbell M. A highly sensitive adenylate cyclase assay. Anal Biochem. 1974 Apr;58(2):541–548. doi: 10.1016/0003-2697(74)90222-x. [DOI] [PubMed] [Google Scholar]
  33. Schramm M., Selinger Z. Message transmission: receptor controlled adenylate cyclase system. Science. 1984 Sep 21;225(4668):1350–1356. doi: 10.1126/science.6147897. [DOI] [PubMed] [Google Scholar]
  34. Seamon K. B., Daly J. W. Forskolin: a unique diterpene activator of cyclic AMP-generating systems. J Cyclic Nucleotide Res. 1981;7(4):201–224. [PubMed] [Google Scholar]
  35. 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]

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