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Expression and characterization of parathyroid hormone receptors on fetal rat calvaria cells in culture

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Abstract

The expression of parathyroid hormone (PTH) receptors during osteoblastic differentiation was studied using long-term cultures of fetal rat calvaria (RC) cells. Scatchard and cross-linking studies with [125I]-labeled [Nle8,18, Tyr34] bovine PTH (1-34) amide indicated that RC cells expressed a single class of saturable, high affinity PTH receptors with a Kd of about 1.0 nM and a molecular mass of about 76 kDa. The number of receptors increased after plating, reached a maximum on day 15 (5.01 × 104 sites/cell) and decreased thereafter (3.82 × 104 sites/cell on day 20), whereas the osteoblastic markers, such as the alkaline phosphatase activity and number of bone nodules increased progressively up to day 20. These results suggest that PTH receptor expression changes during the osteoblastic lineage and that the highest number of receptors occurs at the differentiated stage prior to osteoblast maturation.

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References

  1. Potts JT, Kronenberg HM, Rosenblatt M: Parathyroid hormone. Adv Protein Chem 35: 323–396, 1982

    CAS  PubMed  Google Scholar 

  2. Abou-Samra A-B, Juppner 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 triphosphates and increases intracellular free calcium. Proc Natl Acad Sci USA 89: 2732–2736, 1992

    CAS  PubMed  Google Scholar 

  3. Rouleau MF, Mitchell J, Goltzman D: In vivo distribution of parathyroid hormone receptors in bone: evidence that a predominant osseous target cell is not the mature osteoblast. Endocrinology 123: 187–191, 1988

    CAS  PubMed  Google Scholar 

  4. Heath JK, Rodan SB, Yoon KG et al.: Rat calvarial cell lines immortalized with SV-40 large T-antigen: Costitutive and retinoic acid-inducible expression of osteoblastic features. Endocrinology 124: 3060–3068, 1989

    CAS  PubMed  Google Scholar 

  5. Lee K, Deeds JD, Bond AT et al.: In situ localzation of PTH/PTHrP receptor mRNA in the bone of fetal and young rat. Bone 14: 341–345, 1993

    Article  CAS  PubMed  Google Scholar 

  6. Bellows, CG, Aubin JE, Heersche, JNM et. al.: Mineralized bone nodules formed in vitro from enzymatically released rat calvaria cell populations. Calcif Tissue Int 38: 143–154, 1986

    CAS  PubMed  Google Scholar 

  7. Bellows CG, Aubin JE: Determination of numbers of osteoprogenitors present in isolated fetal rat calvaria cells in vitro. Dev Biol 133: 8–13, 1989

    Article  CAS  PubMed  Google Scholar 

  8. Bellows CG, Ishida H, Aubin JE et al.: Parathyroid hormone reversibly suppresses the differentiation of osteoprogenitor cells into functional osteoblasts. Endocrinology 127: 3111–3116, 1990

    CAS  PubMed  Google Scholar 

  9. Owen TA, Aronow M, Shalhoub V et al.: Progressive development of the rat osteoblast phenotype in vitro: reciplocal relationships in expression of genes associated with osteoblast proliferation and differentiation during formation of the bone extracellular matrix. J dell Physiol 143: 420–430, 1990

    CAS  Google Scholar 

  10. Kasahara S, Nishikawa S, Ishida H et al.: The role of 5′-methylthioadenosine on rat calvaria cell differentiation. Biochem Biophys Res Commun 182: 817–823, 1992

    Article  CAS  PubMed  Google Scholar 

  11. Nagata T, Bellows CG, Kasugai S et al.: Biosynthesis of bone proteins [SPP-1 (secreted phosphoprotein-1, osteopontin), BSP (bone sialoprotein) and SPARC (osteonectin)] in association with mineralized-tissue formation by fetal-rat calvarial cells in culture. Bichem J 274: 513–520, 1991

    CAS  Google Scholar 

  12. Sato K, Han DC, Ozawa M et al.: A highly sensitive bioassay for PTH using ROS17/2.8 subclonal cells. Acta Endocrinal 116: 113–120, 1987

    Google Scholar 

  13. Shigeno C, Hiraki Y, Westerberg DP et al.: Photoaffinity labeling of parathyroid hormone receptors in clonal rat osteosarcoma cells. J Biol Chem 263: 3864–3871, 1988

    CAS  PubMed  Google Scholar 

  14. Lowry OH, Roberts NR, Wu ML et al.: The quantitative histochemistry of brain: Enzyme measurements. J Biol Chem 207: 19–37, 1954

    CAS  PubMed  Google Scholar 

  15. Enomoto M, Kinosita A, Pan HO et al.: Demonstration of receptors for parathyroid hormone cultured rabbit costal chondrocytes. Biochem Biophys Res Commun 162: 1222–1229, 1989

    Article  CAS  PubMed  Google Scholar 

  16. Takigawa M, Kinosita A, Enomoto M et al.: Effects of various growth and differentiation factors on expression of parathyroid hormone receptors on rabbit costal chondrocytes in culture. Endocrinology 129: 868–876, 1991

    CAS  PubMed  Google Scholar 

  17. van der Plas A, Feyen JHM, Nijweide PJ et al.: Direct effect of parathyroid hormone on the proliferation of osteoblast-like cells; a possible involvement of cyclic AMP. Biochem Biophys Res Commun 129: 918–925, 1985

    PubMed  Google Scholar 

  18. Somjen D, Binderman I, Schluter KD et al.: Stimulation by defined parathyroid hormone fragments of cell proliferlation in skeletal cell culture. Biochem J 272: 781–785, 1990

    CAS  PubMed  Google Scholar 

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Yamauchi, N., Ohishi, K., Nishikawa, S. et al. Expression and characterization of parathyroid hormone receptors on fetal rat calvaria cells in culture. J Bone Miner Metab 12 (Suppl 1), S33–S37 (1994). https://doi.org/10.1007/BF02375672

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