Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

Sphingosine-1-phosphate mobilizes osteoclast precursors and regulates bone homeostasis

A Corrigendum to this article was published on 17 June 2010

Abstract

Osteoclasts are the only somatic cells with bone-resorbing capacity and, as such, they have a critical role not only in normal bone homeostasis (called ‘bone remodelling’) but also in the pathogenesis of bone destructive disorders such as rheumatoid arthritis and osteoporosis1. A major focus of research in the field has been on gene regulation by osteoclastogenic cytokines such as receptor activator of NF-κB-ligand (RANKL, also known as TNFSF11) and TNF-α, both of which have been well documented to contribute to osteoclast terminal differentiation2,3. A crucial process that has been less well studied is the trafficking of osteoclast precursors to and from the bone surface, where they undergo cell fusion to form the fully differentiated multinucleated cells that mediate bone resorption. Here we report that sphingosine-1-phosphate (S1P), a lipid mediator enriched in blood4,5, induces chemotaxis and regulates the migration of osteoclast precursors not only in culture but also in vivo, contributing to the dynamic control of bone mineral homeostasis. Cells with the properties of osteoclast precursors express functional S1P1 receptors and exhibit positive chemotaxis along an S1P gradient in vitro. Intravital two-photon imaging of bone tissues showed that a potent S1P1 agonist, SEW2871, stimulated motility of osteoclast precursor-containing monocytoid populations in vivo. Osteoclast/monocyte (CD11b, also known as ITGAM) lineage-specific conditional S1P1 knockout mice showed osteoporotic changes due to increased osteoclast attachment to the bone surface. Furthermore, treatment with the S1P1 agonist FTY720 relieved ovariectomy-induced osteoporosis in mice by reducing the number of mature osteoclasts attached to the bone surface. Together, these data provide evidence that S1P controls the migratory behaviour of osteoclast precursors, dynamically regulating bone mineral homeostasis, and identifies a critical control point in osteoclastogenesis that may have potential as a therapeutic target.

This is a preview of subscription content, access via your institution

Relevant articles

Open Access articles citing this article.

Access options

Buy article

Get time limited or full article access on ReadCube.

$32.00

All prices are NET prices.

Figure 1: Expression and function of S1P receptors in osteoclast precursor monocytes.
Figure 2: In vivo S1P-mediated increase in motility of osteoclast precursor monocytes visualized using intravital two-photon imaging.
Figure 3: In vivo affect of S1P 1 on bone mineral metabolism.
Figure 4: Preventive effect of FTY720 on ovariectomy-induced osteoporosis.

References

  1. Teitelbaum, S. L. Bone resorption by osteoclasts. Science 289, 1504–1508 (2000)

    ADS  CAS  Article  Google Scholar 

  2. Teitelbaum, S. L. & Ross, F. P. Genetic regulation of osteoclast development and function. Nature Rev. Genet. 4, 638–649 (2003)

    CAS  Article  Google Scholar 

  3. Karsenty, G. & Wagner, E. F. Reaching a genetic and molecular understanding of skeletal development. Dev. Cell 2, 389–406 (2002)

    CAS  Article  Google Scholar 

  4. Rosen, H. & Goetzl, E. J. Sphingosine 1-phosphate and its receptors: an autocrine and paracrine network. Nature Rev. Immunol. 5, 560–570 (2005)

    CAS  Article  Google Scholar 

  5. Cyster, J. G. Chemokines, sphingosine-1-phosphate, and cell migration in secondary lymphoid organs. Annu. Rev. Immunol. 23, 127–159 (2005)

    CAS  Article  Google Scholar 

  6. Bokoch, G. M., Katada, T., Northup, J. K., Ui, M. & Gilman, A. G. Purification and properties of the inhibitory guanine nucleotide-binding regulatory component of adenylate cyclase. J. Biol. Chem. 259, 3560–3567 (1984)

    CAS  PubMed  Google Scholar 

  7. Akbar, H., Cancelas, J., Williams, D. A., Zheng, J. & Zheng, Y. Rational design and applications of a Rac GTPase-specific small molecule inhibitor. Methods Enzymol. 406, 554–565 (2006)

    CAS  Article  Google Scholar 

  8. Spiegel, S., English, D. & Milstien, S. Sphingosine 1-phosphate signaling: providing cells with a sense of direction. Trends Cell Biol. 12, 236–242 (2002)

    CAS  Article  Google Scholar 

  9. Takuwa, Y. Subtype-specific differential regulation of Rho family G proteins and cell migration by the Edg family sphingosine-1-phosphate receptors. Biochim. Biophys. Acta 1582, 112–120 (2002)

    CAS  Article  Google Scholar 

  10. Mazo, I. B. et al. Hematopoietic progenitor cell rolling in bone marrow microvessels: parallel contributions by endothelial selectins and vascular cell adhesion molecule 1. J. Exp. Med. 188, 465–474 (1998)

    CAS  Article  Google Scholar 

  11. Mazo, I. B. et al. Bone marrow is a major reservoir and site of recruitment for central memory CD8+ T cells. Immunity 22, 259–270 (2005)

    CAS  Article  Google Scholar 

  12. Jung, S. et al. Analysis of fractalkine receptor CX3CR1 function by targeted deletion and green fluorescent protein reporter gene insertion. Mol. Cell. Biol. 20, 4106–4114 (2000)

    CAS  Article  Google Scholar 

  13. Niess, J. H. et al. CX3CR1-mediated dendritic cell access to the intestinal lumen and bacterial clearance. Science 307, 254–258 (2005)

    ADS  CAS  Article  Google Scholar 

  14. Burnett, S. H. et al. Conditional macrophage ablation in transgenic mice expressing a Fas-based suicide gene. J. Leukoc. Biol. 75, 612–623 (2004)

    CAS  Article  Google Scholar 

  15. Sanna, M. G. et al. Sphingosine 1-phosphate (S1P) receptor subtypes S1P1 and S1P3, respectively, regulate lymphocyte recirculation and heart rate. J. Biol. Chem. 279, 13839–13848 (2004)

    CAS  Article  Google Scholar 

  16. Wei, S. H. et al. Sphingosine 1-phosphate type 1 receptor agonism inhibits transendothelial migration of medullary T cells to lymphatic sinuses. Nature Immunol. 6, 1228–1235 (2005)

    CAS  Article  Google Scholar 

  17. Liu, Y. et al. Edg-1, the G protein-coupled receptor for sphingosine-1-phoshate, is essential for vascular maturation. J. Clin. Invest. 106, 951–961 (2000)

    CAS  Article  Google Scholar 

  18. Allende, M. L. et al. G-protein-coupled receptor S1P1 acts within endothelial cells to regulate vascular maturation. Blood 102, 3665–3667 (2003)

    CAS  Article  Google Scholar 

  19. Ferron, M. & Vacher, J. Targeted expression of Cre recombinase in macrophages and osteoclasts in transgenic mice. Genesis 41, 138–145 (2005)

    CAS  Article  Google Scholar 

  20. Filgueira, L. Fluorescence-based staining for tartrate-resistant acidic phosphatase (TRAP) in osteoclasts combined with other fluorescent dyes and protocols. J. Histochem. Cytochem. 52, 411–414 (2004)

    CAS  Article  Google Scholar 

  21. Stoller, P., Reiser, K. M., Celliers, P. M. & Rubenchik, A. M. Polarization-modulated second harmonic generation in collagen. Biophys. J. 82, 3330–3342 (2002)

    CAS  Article  Google Scholar 

  22. Yu, X., Huang, Y., Collin-Osdoby, P. & Osdoby, P. Stromal cell-derived factor-1 (SDF-1) recruits osteoclast precursors by inducing chemotaxis, matrix metalloprotease-9 (MMP-9) activity, and collagen transmigration. J. Bone Miner. Res. 18, 1404–1418 (2003)

    CAS  Article  Google Scholar 

  23. Katayama, Y. et al. Signals from the sympathetic nervous system regulate hematopoietic stem cell egress from bone marrow. Cell 124, 407–421 (2006)

    CAS  Article  Google Scholar 

  24. Matloubian, M. et al. Lymphocyte egress from thymus and peripheral lymphoid organs is dependent on S1P receptor 1. Nature 427, 355–360 (2004)

    ADS  CAS  Article  Google Scholar 

  25. Cyster, J. G. Chemokines, sphingosine-1-phosphate, and cell migration in secondary lymphoid organs. Annu. Rev. Immunol. 23, 127–159 (2005)

    CAS  Article  Google Scholar 

  26. Kappos, L. et al. Oral fingolimod (FTY720) for relapsing multiple sclerosis. N. Engl. J. Med. 355, 1124–1140 (2006)

    CAS  Article  Google Scholar 

  27. Tedesco-Silva, H. et al. FTY720 versus mycophenolate mofetil in de novo renal transplantation: six-month results of a double-blind study. Transplantation 84, 885–892 (2007)

    CAS  Article  Google Scholar 

  28. Kobayashi, K. et al. Tumor necrosis factor α stimulates osteoclast differentiation by a mechanism independent of the ODF/RANKL-RANK interaction. J. Exp. Med. 191, 275–286 (2000)

    CAS  Article  Google Scholar 

  29. Ishii, M. et al. RANKL-induced expression of tetraspanin CD9 in lipid raft membrane microdomain is essential for cell fusion during osteoclastogenesis. J. Bone Miner. Res. 21, 965–976 (2006)

    CAS  Article  Google Scholar 

  30. Okamoto, H. et al. Inhibitory regulation of Rac activation, membrane ruffling, and cell migration by the G protein-coupled sphingosine-1-phosphate receptor EDG5 but not EDG1 or EDG3. Mol. Cell. Biol. 20, 9247–9261 (2000)

    CAS  Article  Google Scholar 

Download references

Acknowledgements

We thank U. H. von Andrian and I. B. Mazo for their help with the technique of intravital skull bone imaging. We also thank Y. Takuwa and N. Sugimoto for discussions, and P. M. Murphy and S.Venkatesan for their help in imaging in vitro chemotaxis using the EZ-Taxiscan. This work was supported in part by the Intramural Research Program of the National Institute of Allergy and Infectious Diseases, NIH, US Department of Health and Human Services, and by a fellowship grant to M.I. from the International Human Frontier Science Program.

Author Contributions M.I. performed most of experiments, with the assistance of J.G.E. and Y.S. for two-photon microscopy and for the in vitro osteoclast culture system, respectively. F.K. developed the unsupervised segmentation software and performed the computational analyses used to quantify the osteoclast-bone surface interface, with the assistance of M.M.-S. J.V. and R.L.P. generated CD11b-Cre transgenic and S1PR1loxP knock-in mice, respectively. R.N.G. helped M.I. in designing and interpreting experiments, as well as in writing the paper.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Ronald N. Germain.

Supplementary information

Supplementary Information

This file contains Supplementary Figures 1-10 with Legends, Supplementary Tables 1-4, Supplementary Video Legends 1-7 and Supplementary References. (PDF 1213 kb)

Supplementary Video 1

This file shows in vitro chemotaxis of RAW264.7 cells in control conditions, detected using the EZ-Taxiscan device. Images were taken every minute for 2 hours. Scale bars represent 150 µm. (See legend in file s1). (MOV 1477 kb)

Supplementary Video 2

This file shows in vitro chemotaxis of RAW264.7 cells toward S1P gradient (10-8 M), detected using the EZ-Taxiscan device. Images were taken every minute for 2 hours. Scale bars represent 150 µm. (See legend in file s1). (MOV 1981 kb)

Supplementary Video 3

This file shows intravital two-photon imaging of mouse skull bone tissues of CX3CR1-EGFP knock-in (heterozygous) mice before application of SEW2871. Scale bars represent 50 µm. (See legend in file s1). (MOV 1073 kb)

Supplementary Video 4

This file shows intravital two-photon imaging of mouse skull bone tissues of CX3CR1-EGFP knock-in (heterozygous) mice 30 minutes after application of SEW2871. Scale bars represent 50 µm. (See legend in file s1). (MOV 1601 kb)

Supplementary Video 5

This file shows intravital two-photon imaging of mouse skull bone tissues of CSF1R-EGFP transgenic mice before application of SEW2871. Scale bars represent 50 µm. (See legend in file s1). (MOV 1299 kb)

Supplementary Video 6

This file shows intravital two-photon imaging of mouse skull bone tissues of CSF1R-EGFP transgenic mice 30 minutes after application of SEW2871. Scale bars represent 50 µm. (See legend in file s1). (MOV 1560 kb)

Supplementary Video 7

This file shows intravital two-photon imaging of mouse skull bone tissues of CX3CR1-EGFP knock-in (heterozygous) mice, pre-treated with FTY720 (3 mg/kg, i.p.) 4 hours before imaging. Scale bars represent 50 µm. (See legend in file s1). (MOV 988 kb)

PowerPoint slides

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Ishii, M., Egen, J., Klauschen, F. et al. Sphingosine-1-phosphate mobilizes osteoclast precursors and regulates bone homeostasis. Nature 458, 524–528 (2009). https://doi.org/10.1038/nature07713

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nature07713

Further reading

Comments

By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.

Search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing