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Treatment of Myoblastic C2C12 Cells with BMP-2 Stimulates Vitamin D-induced Formation of Osteoclasts

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Abstract

The interaction of osteoclast precursors with osteoblasts and/or stromal cells is essential for the formation of mature osteoclasts and the resorption of bone. We found that myoblastic C2C12 cells induced the differentiation of mouse spleen cells into tartrate-resistant acid phosphatase-positive (TRAP-positive) multinucleated cells in the presence of 10−7 M 1α,25-dihydroxyvitamin D3 [1α,25(OH)2D3] and that C2C12 cells that had been treated with bone morphogenetic protein-2 (BMP-2) dose-dependently stimulated the formation of osteoclasts. The newly developed TRAP-positive multinucleated cells were capable of resorbing mineralized tissues. Treatment of C2C12 cells with BMP-2 for 24 h enhanced the subsequent expression in C2C12 cells of mRNA for the receptor activator of nuclear factor-κB ligand (RANKL) in the presence of 1α,25(OH)2D3. Since the formation of osteoclasts was inhibited dose-dependently by exogenous OPG, the expression of RANKL in response to BMP-2 appeared to be critical for the formation of osteoclasts. Our findings suggest that BMP-2 might play an important role in the differentiation of cells that support osteoclastogenesis.

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References

  1. AM Parfitt (1994) ArticleTitleOsteonal and hemi-osteonal remodeling: the spatial and temporal framework for signal traffic in adult human bone. J Cell Biochem 55 273–286 Occurrence Handle1:STN:280:ByqD2M7gslQ%3D Occurrence Handle7962158

    CAS  PubMed  Google Scholar 

  2. SC Manolagas RL Jilka (1995) ArticleTitleBone marrow, cytokines, and bone remodeling. Emerging insights into the pathophysiology of osteoporosis. N Engl J Med 332 305–311 Occurrence Handle1:STN:280:ByqC3MrntF0%3D Occurrence Handle7816067

    CAS  PubMed  Google Scholar 

  3. SL Teitelbaum (2000) ArticleTitleBone resorption by osteoclasts. Science 289 1504–1508 Occurrence Handle10.1126/science.289.5484.1504 Occurrence Handle10968780

    Article  PubMed  Google Scholar 

  4. N Udagawa N Takahashi T Akatsu H Tanaka T Sasaki T Nishihara T Koga TJ Martin T Suda (1990) ArticleTitleOrigin of osteoclasts: mature monocytes and macrophages are capable of differentiating into osteoclasts under a suitable microenvironment prepared by bone marrow-derived stromal cells. Proc Natl Acad Sci USA 87 7260–7264 Occurrence Handle1:STN:280:By%2BA1cvpvVM%3D Occurrence Handle2169622

    CAS  PubMed  Google Scholar 

  5. N Takahashi T Akatsu N Udagawa T Sasaki A Yamaguchi JM Moseley TJ Martin T Suda (1988) ArticleTitleOsteoblastic cells are involved in osteoclast formation. Endocrinology 123 2600–2602 Occurrence Handle1:CAS:528:DyaL1cXmt1aquro%3D Occurrence Handle2844518

    CAS  PubMed  Google Scholar 

  6. T Suda I Nakamura E Jimi N Takahashi (1997) ArticleTitleRegulation of osteoclast function. J Bone Miner Res 12 869–879 Occurrence Handle1:CAS:528:DyaK2sXjs1yltr0%3D Occurrence Handle9169344

    CAS  PubMed  Google Scholar 

  7. N Takahashi N Udagawa T Suda (1999) ArticleTitleA new member of tumor necrosis factor ligand family, ODF/OPGL/TRANCE/RANKL, regulates osteoclast differentiation and function. Biochem Biophys Res Commun 256 449–455 Occurrence Handle10.1006/bbrc.1999.0252 Occurrence Handle10080918

    Article  PubMed  Google Scholar 

  8. DM Anderson E Maraskovsky WL Billingsley WC Dougall ME Tometsko ER Roux MC Teepe RF DuBose D Cosman L Galibert (1997) ArticleTitleA homologue of the TNF receptor and its ligand enhance T-cell growth and dendritic-cell function. Nature 390 175–179 Occurrence Handle9367155

    PubMed  Google Scholar 

  9. BR Wong J Rho J Arron E Robinson J Orlinick M Chao S Kalachikov E Cayani FS Bartlett III WN Frankel SY Lee Y Choi (1997) ArticleTitleTRANCE is a novel ligand of the tumor necrosis factor receptor family that activates c-Jun N-terminal kinase in T cells. J Biol Chem 272 25190–25194 Occurrence Handle1:CAS:528:DyaK2sXms1ehsbo%3D Occurrence Handle9312132

    CAS  PubMed  Google Scholar 

  10. H Yasuda N Shima N Nakagawa K Yamaguchi M Kinosaki S Mochizuki A Tomoyasu K Yano M Goto A Murakami E Tsuda T Morinaga K Higashio N Udagawa N Takahashi T Suda (1998) ArticleTitleOsteoclast differentiation factor is a ligand for osteoprotegerin/osteoclastogenesis-inhibitory factor and is identical to TRANCE/RANKL. Proc Natl Acad Sci USA 95 3597–3602 Occurrence Handle1:CAS:528:DyaK1cXitlKjsLY%3D Occurrence Handle9520411

    CAS  PubMed  Google Scholar 

  11. DL Lacey E Timms HL Tan et al. (1998) ArticleTitleOsteoprotegerin ligand is a cytokine that regulates osteoclast differentiation and activation. Cell 93 165–176 Occurrence Handle9568710

    PubMed  Google Scholar 

  12. H Hsu DL Lacey CR Dunstan et al. (1999) ArticleTitleTumor necrosis factor receptor family member RANK mediates osteoclast differentiation and activation induced by osteoprotegerin ligand. Proc Natl Acad Sci USA 96 3540–3545 Occurrence Handle1:CAS:528:DyaK1MXjslCitL8%3D Occurrence Handle10097072

    CAS  PubMed  Google Scholar 

  13. N Nakagawa M Kinosaki K Yamaguchi et al. (1998) ArticleTitleRANK is the essential signaling receptor for osteoclast differentiation factor in osteoclastogenesis. Biochem Biophys Res Commun 253 395–400 Occurrence Handle10.1006/bbrc.1998.9788 Occurrence Handle9878548

    Article  PubMed  Google Scholar 

  14. E Jimi I Nakamura H Amano et al. (1996) ArticleTitleOsteoclast function is activated by osteoblastic cells through a mechanism involving cell-to-cell contact. Endocrinology 137 2187–2190

    Google Scholar 

  15. YY Kong H Yoshida I Sarosi et al. (1999) ArticleTitleOPGL is a key regulator of osteoclastogenesis, lymphocyte development and lymph-node organogenesis. Nature 397 315–323 Occurrence Handle1:CAS:528:DyaK1MXhtVOqs7w%3D Occurrence Handle9950424

    CAS  PubMed  Google Scholar 

  16. WC Dougall M Glaccum K Charrier et al. (1999) ArticleTitleRANK is essential for osteoclast and lymph node development. Genes Dev 13 2412–2424 Occurrence Handle1:CAS:528:DyaK1MXmsVCgtr4%3D Occurrence Handle10500098

    CAS  PubMed  Google Scholar 

  17. WS Simonet DL Lacey CR Dunstan et al. (1997) ArticleTitleOsteoprotegerin: a novel secreted protein involved in the regulation of bone density. Cell 89 309–319 Occurrence Handle9108485

    PubMed  Google Scholar 

  18. H Yasuda N Shima N Nakagawa et al. (1998) ArticleTitleIdentity of osteoclastogenesis inhibitory factor (OCIF) and osteoprotegerin (OPG): a mechanism by which OPG/OCIF inhibits osteoclastogenesis in vitro. Endocrinology 139 1329–1337 Occurrence Handle9492069

    PubMed  Google Scholar 

  19. A Mizuno N Amizuka K Irie et al. (1998) ArticleTitleSevere osteoporosis in mice lacking osteoclastogenesis inhibitory factor/osteoprotegerin. Biochem Biophys Res Commun 247 610–615

    Google Scholar 

  20. N Bucay I Sarosi CR Dunstan et al. (1998) ArticleTitleOsteoprotegerin-deficient mice develop early onset osteoporosis and arterial calcification. Genes Dev 12 1260–1268 Occurrence Handle1:CAS:528:DyaK1cXjtVejsrs%3D Occurrence Handle9573043

    CAS  PubMed  Google Scholar 

  21. HM Blau CP Chiu C Webster (1983) ArticleTitleCytoplasmic activation of human nuclear genes in stable heterocaryons. Cell 32 1171–1180 Occurrence Handle1:CAS:528:DyaL3sXhvFGls74%3D Occurrence Handle6839359

    CAS  PubMed  Google Scholar 

  22. T Katagiri A Yamaguchi M Komaki et al. (1994) ArticleTitleBone morphogenetic protein-2 converts the differentiation pathway of C2C12 myoblasts into the osteoblast lineage. J Cell Biol 127 1755–1766 Occurrence Handle1:CAS:528:DyaK2cXmvFemtLs%3D Occurrence Handle7798324

    CAS  PubMed  Google Scholar 

  23. M Fujii K Takeda T Imamura et al. (1999) ArticleTitleRoles of bone morphogenetic protein type I receptors and Smad proteins in osteoblast and chondroblast differentiation. Mol Biol Cell 10 3801–3813 Occurrence Handle1:CAS:528:DyaK1MXntlyrsLg%3D Occurrence Handle10564272

    CAS  PubMed  Google Scholar 

  24. P Ducy R Zhang V Geoffroy AL Ridall G Karsenty (1997) ArticleTitleOsf2/Cbfa1: a transcriptional activator of osteoblast differentiation. Cell 89 747–754 Occurrence Handle1:CAS:528:DyaK2sXjs1ejsbg%3D Occurrence Handle9182762

    CAS  PubMed  Google Scholar 

  25. MH Lee A Javed HJ Kim et al. (1999) ArticleTitleTransient upregulation of CBFA1 in response to bone morphogenetic protein-2 and transforming growth factor beta1 in C2C12 myogenic cells coincides with suppression of the myogenic phenotype but is not sufficient for osteoblast differentiation. J Cell Biochem 73 114–125 Occurrence Handle10.1002/(SICI)1097-4644(19990401)73:1<114::AID-JCB13>3.3.CO;2-D Occurrence Handle1:CAS:528:DyaK1MXht12lu70%3D Occurrence Handle10088730

    Article  CAS  PubMed  Google Scholar 

  26. N Udagawa N Takahashi T Akatsu et al. (1989) ArticleTitleThe bone marrow-derived stromal cell lines MC3T3-G2/PA6 and ST2 support osteoclast-like cell differentiation in cocultures with mouse spleen cells. Endocrinology 125 1805–1813 Occurrence Handle1:CAS:528:DyaL1MXmtV2lu7c%3D Occurrence Handle2676473

    CAS  PubMed  Google Scholar 

  27. H Kaneko T Arakawa H Mano et al. (2000) ArticleTitleDirect stimulation of osteoclastic bone resorption by bone morphogenetic protein (BMP)-2 and expression of BMP receptors in mature osteoclasts. Bone 27 479–486 Occurrence Handle10.1016/S8756-3282(00)00358-6 Occurrence Handle1:CAS:528:DC%2BD3cXnt1Sjtrg%3D Occurrence Handle11033442

    Article  CAS  PubMed  Google Scholar 

  28. K Itoh N Udagawa T Katagiri et al. (2001) ArticleTitleBone morphogenetic protein 2 stimulates osteoclast differentiation and survival supported by receptor activator of nuclear factor-kappaB ligand. Endocrinology 142 3656–3662 Occurrence Handle1:CAS:528:DC%2BD3MXls1SqtLw%3D Occurrence Handle11459815

    CAS  PubMed  Google Scholar 

  29. E Abe M Yamamoto Y Taguchi et al. (2000) ArticleTitleEssential requirement of BMPs-2/4 for both osteoblast and osteoclast formation in murine bone marrow cultures from adult mice: antagonism by noggin. J Bone Miner Res 15 663–673 Occurrence Handle1:CAS:528:DC%2BD3cXisFWmtrg%3D Occurrence Handle10780858

    CAS  PubMed  Google Scholar 

  30. LC Hofbauer CR Dunstan TC Spelsberg BL Riggs S Khosla (1998) ArticleTitleOsteoprotegerin production by human osteoblast lineage cells is stimulated by vitamin D, bone morphogenetic protein-2, and cytokines. Biochem Biophys Res Commun 250 776–781

    Google Scholar 

  31. M Wan X Shi X Feng X Cao (2001) ArticleTitleTranscriptional mechanisms of bone morphogenetic protein-induced Osteoprotegerin gene expression. J Biol Chem 276 10119–10125 Occurrence Handle10.1074/jbc.M006918200 Occurrence Handle1:CAS:528:DC%2BD3MXis1Oksbg%3D Occurrence Handle11139569

    Article  CAS  PubMed  Google Scholar 

  32. K Thirunavukkarasu DL Halladay RR Miles et al. (2000) ArticleTitleThe osteoblast-specific transcription factor Cbfa1 contributes to the expression of osteoprotegerin, a potent inhibitor of osteoclast differentiation and function. J Biol Chem 275 25163–25172 Occurrence Handle1:CAS:528:DC%2BD3cXmtVeiuro%3D Occurrence Handle10833509

    CAS  PubMed  Google Scholar 

  33. SC Bae KS Lee YW Zhang Y Ito (2001) ArticleTitleIntimate relationship between TGF-β/BMP signaling and runt domain transcription factor, PEBP2/CBF. J Bone Joint Surg Am 83-A S48–S55 Occurrence Handle11263665

    PubMed  Google Scholar 

  34. S Gallea F Lallemand A Atfi et al. (2001) ArticleTitleActivation of mitogen-activated protein kinase cascades is involved in regulation of bone morphogenetic protein-2-induced osteoblast differentiation in pluripotent C2C12 cells. Bone 28 491–498 Occurrence Handle10.1016/S8756-3282(01)00415-X Occurrence Handle1:CAS:528:DC%2BD3MXjtlChsL0%3D Occurrence Handle11344048

    Article  CAS  PubMed  Google Scholar 

  35. JM Graff A Bansal DA Melton (1996) ArticleTitleXenopus Mad proteins transduce distinct subsets of signals for the TGFβ superfamily. Cell 85 479–487 Occurrence Handle1:CAS:528:DyaK28XjtFyitrY%3D Occurrence Handle8653784

    CAS  PubMed  Google Scholar 

  36. PA Hoodless T Haerry S Abdollah et al. (1996) ArticleTitleMADR1, a MAD-related protein that functions in BMP2 signaling pathways. Cell 85 489–500 Occurrence Handle1:CAS:528:DyaK28XjtFyitrc%3D Occurrence Handle8653785

    CAS  PubMed  Google Scholar 

  37. F Liu A Hata JC Baker J Doody J Carcamo RM Harland J Massague (1996) ArticleTitleA human Mad protein acting as a BMP-regulated transcriptional activator. Nature 381 561–562 Occurrence Handle10.1038/381561a0 Occurrence Handle8637588

    Article  PubMed  Google Scholar 

  38. GH Thomsen () ArticleTitleXenopus mothers against decapentaplegic is an embryonic ventralizing agent that acts downstream of the BMP-2/4 receptor. Development 122 2359–2366

    Google Scholar 

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Acknowledgements

The authors thank Drs. A. Yamaguchi (Nagasaki University) and T. Katagiri (Showa University) for their helpful comments. This work was supported by Grants-in-Aid for Scientific Research from the Ministry of Education, Science, Sports and Culture of Japan, and by grants from the Uehara Memorial Foundation and the Smoking Research Foundation.

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  1. Department of Biological Sciences, Tokyo Institute of Technology, 4259-Nagatsuta-cho, Midori-ku, Yokohama 226-8501, Japan

    E. Otsuka, M. Notoya & H. Hagiwara

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  1. E. Otsuka
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  2. M. Notoya
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  3. H. Hagiwara
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Correspondence to H. Hagiwara.

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Otsuka, E., Notoya, M. & Hagiwara, H. Treatment of Myoblastic C2C12 Cells with BMP-2 Stimulates Vitamin D-induced Formation of Osteoclasts . Calcif Tissue Int 73, 72–77 (2003). https://doi.org/10.1007/s00223-002-1071-0

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