Skip to main content

Advertisement

SpringerLink
Log in

Differential regulation of the PTH/PTHrP receptor and its mRNA in human osteoblast-like SaOS-2 cells

  • Published:
Journal of Bone and Mineral Metabolism Aims and scope Submit manuscript

Abstract

Using wild-type SaOS-2 cells and a mutant cAMP-resistant subclone Ca#4A, we have recently shown that homologous down-regulation of PTH/PTHrP receptor (PTH/PTHrPR) is mediated by mechanisms independent of PKA activation, and that homologous down-regulation of PTH/PTHrPR mRNA occurs later than the decline in functional cell surface receptors via a mechanism that does not involve enhanced mRNA degradation or new protein synthesis but does depend on cAMP/PKA. Treatment of SaOS-2 cells with [Arg2]hPTH(1-34) or hPTH(3-34) for 24 h, both of which bind to the human PTH/PTHrPR as well as hPTH(1-34) but fail to stimulate cAMP/PKA efficiently, does not cause down-regulation of PTH/PTHrPR mRNA; however, the same treatment did down-regulate PTH/PTHrPR protein. Incubation with isoproterenol for 24 h, which activates both PKA andβ-adrenergic receptor kinase (βARK) in these cells, induced significant down-regulation of both the PTH/PTHrPR and its mRNA. However, treatment with vasoactive intestinal peptide (VIP) or PGE2 for 24 h, both of which stimulate the cAMP/PKA pathway in these cells, did not down-regulate PTH/PTHrPR mRNA. We conclude that both PKA-dependent and -independent (such asβARK) mechanisms are required for regulation of the PTH/PTHrPR and its mRNA.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. Abou-Samra A-B, Jüppner H, Potts Jr JT et al.: Inactivation of pertussis toxin-sensitive guanyl nucleotide-binding proteins increase parathyroid hormone receptors and reverse agonist-induced receptor down-regulation in ROS 17/2.8 cells. Endocrinology 125: 2594–2599, 1989

    CAS  PubMed  Google Scholar 

  2. Abou-Samra A-B, Zajac JD, Schiffer-Alberts D et al.: Cyclic adenosine 3′,5′-monophosphate (cAMP)-dependent and cAMP-independent regulation of parathyroid hormone receptors in UMR 106-01 osteoblastic osteosarcoma cells. Endocrinology 129: 2547–2554, 1991

    CAS  PubMed  Google Scholar 

  3. Fukayama S, Tashjian Jr AH, Bringhurst FR: Mechanisms of desensitization to parathyroid hormone in human osteoblast-like SaOS-2 cells. Endocrinology 131: 1757–1769, 1992

    CAS  PubMed  Google Scholar 

  4. Sugimoto T, Ikeda K, Kano J, et al.: Cross-talk of parathyroid hormone-responsive dual signal transduction systems in osteoblastic osteosarcoma cells: its role in PTH-induced homologous desensitization of intracellular calcium response. J Cell Physiol 158: 374–380, 1994

    Article  CAS  PubMed  Google Scholar 

  5. Hohmann EL, Tashjian Jr AH: Functional receptors for vasoactive intestinal peptide on human osteosarcoma cells. Endocrinology 114: 1321–1327, 1984

    CAS  PubMed  Google Scholar 

  6. Fukayama S, Kearns AK, Skurat RM et al.: Protein kinase A-dependent inhibition of alkaline phosphatase release by SaOS-2 human osteoblastic cells: studies in new mutant cell lines that express a cyclic AMP-resistant phenotype. Cell Regul 2: 889–896, 1991

    CAS  PubMed  Google Scholar 

  7. Fukayama S, Schipani E, Jüppner H et al.: Role of protein kinase-A in homologous down-regulation of parathyroid hormone (PTH)/PTH-related peptide receptor messenger ribonucleic acid in human osteoblast-like SaOS-2 cells. Endocrinology 134: 1851–1858, 1994

    Article  CAS  PubMed  Google Scholar 

  8. Abou-Samra A-B, Jüppner H, Force T et al.: Expression cloning of a common receptor for parathyroid hormone and parathyroid hormone-related peptide from rat osteoblast-like cells: a single receptor stimulates intracellular accumulation of both cAMP and inositol trisphosphates and increases intracellular free calcium. Proc Natl Acad Sci USA 89: 2732–2736, 1992

    CAS  PubMed  Google Scholar 

  9. Bringhurst FR, Jüppner H, Guo J et al.: Cloned, stably expressed parathyroid hormone (PTH)/PTH-related peptide receptors activate multiple messenger signals and biological responses in LLC-PK1 kidney cells. Endocrinology 132: 2090–2098, 1993

    Article  CAS  PubMed  Google Scholar 

  10. Schneider H, Feyen JHM, Seuwen K et al.: Cloning and functional expression of human parathyroid hormone receptor. Eur J Pharmacol 246: 149–155, 1993

    CAS  PubMed  Google Scholar 

  11. Boland CJ, Fried RM, Tashjian Jr AH: Measurement of cytosolic free Ca2+ concentrations in human and rat osteosarcoma cells: actions of bone resorption-stimulating hormones. Endocrinology 118: 980–989, 1986

    CAS  PubMed  Google Scholar 

  12. Krug E, Tashjian Jr AH: Time-dependent changes in protein kinase C distribution and disappearance in phorbol-ester-treated human osteosarcoma cells. Cancer Res 47: 2243–2246, 1987

    CAS  PubMed  Google Scholar 

  13. Fukayama S, Tashjian Jr AH: Stimulation by parathyroid hormone of45Ca2+ uptake in osteoblast-like cells: possible involvement of alkaline phosphatase. Endocrinology 126: 1941–1949, 1990

    CAS  PubMed  Google Scholar 

  14. Ofori-Darko E, Tashjian Jr AH: Regulation of pHi in SaOS-2 cells by thrombin: roles of proteolytic activity and cytosolic calcium transients. Am J Physiol 263 (Cell Physiol 32): C1266-C1273, 1992

    CAS  PubMed  Google Scholar 

  15. Moore RE, Smith CK II, Bailey CS et al.: Characterization of beta-adrenergic receptors on rat and human osteoblast-like cells and demonstration that beta-receptor agonists can stimulate bone resorption in organ culture. Bone Miner 23: 301–315, 1993

    CAS  PubMed  Google Scholar 

  16. Sutherland MK, Rao LG, Wylie JN et al.: Carboxylterminal parathyroid hormone peptide (53–84) elevates alkaline phosphatase and osteocalcin mRNA levels in SaOS-2 cells. J Bone Miner Res 9: 453–458, 1994

    CAS  PubMed  Google Scholar 

  17. Iyenger R: Molecular and functional diversity of mammalian Gs-stimulated adenylyl cyclases. FASEB J 7: 768–775, 1993

    Google Scholar 

  18. Hagmann J: Developmentally regulated compartmentalization of adenylate cyclase inDictyostelium discoideum. J Cell Biochem 37: 359–370, 1988

    Article  CAS  PubMed  Google Scholar 

  19. Moger WH: Evidence for compartmentalization of adenosine 3′,5′-monophosphate (cAMP)-dependent protein kinase in rat Leydig cells using site-selective cAMP analogs. Endocrinology 128: 1414–1418, 1991

    CAS  PubMed  Google Scholar 

  20. Yu SS, Lefkowitz RJ, Hausdorff WP:β-adrenergic receptor sequestration. A potential mechanism of receptor resensitization. J Biol Chem 268: 337–341, 1993

    CAS  PubMed  Google Scholar 

  21. Liggett SB, Freedman NJ, Schwinn DA et al.: Structural basis for receptor subtype-specific regulation revealed by a chimericβ 3/β 2-adrenergic receptor. Proc Natl Acad Sci USA 90: 3665–3669, 1993

    CAS  PubMed  Google Scholar 

  22. Barak L, Tiberi M, Freedmann NJ et al.: A highly conserved tyrosine residue in G protein-coupled receptors is required for agonist-mediatedβ 2-adrenergic receptor sequestration. J Biol Chem 269: 2790–2795, 1994

    CAS  PubMed  Google Scholar 

  23. Blind E, Fowlkes ME, Pratt S et al.: Phosphorylation of the C-terminal tail of the PTH/PTHrP receptor by protein kinase C. J Bone Miner Res (Suppl 1) 8: S132, 1993 (Abstract)

Download references

Author information

Authors and Affiliations

  1. Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, MA, U.S.A.

    Shoichi Fukayama Ph.D. & Armen H. Tashijian Jr

  2. Endocrine Unit, Massachusetts General Hospital, and Department of Medicine, Harvard Medical School, Boston, MA, U.S.A.

    F. Richard Bringhurst

  3. Department of Molecular and Cellular Toxicology, Harvard School of Public Health, 665 Huntington Avenue, 02115, Boston, MA, U.S.A.

    Shoichi Fukayama Ph.D. & Armen H. Tashijian Jr

Authors
  1. Shoichi Fukayama Ph.D.
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Armen H. Tashijian Jr
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. F. Richard Bringhurst
    View author publications

    You can also search for this author in PubMed Google Scholar

About this article

Cite this article

Fukayama, S., Tashijian, A.H. & Bringhurst, F.R. Differential regulation of the PTH/PTHrP receptor and its mRNA in human osteoblast-like SaOS-2 cells. J Bone Miner Metab 12 (Suppl 1), S175–S181 (1994). https://doi.org/10.1007/BF02375699

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF02375699

Key words

Search

Navigation