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Matrix-embedded cells control osteoclast formation

Abstract

Osteoclasts resorb the mineralized matrices formed by chondrocytes or osteoblasts. The cytokine receptor activator of nuclear factor-κB ligand (RANKL) is essential for osteoclast formation and thought to be supplied by osteoblasts or their precursors, thereby linking bone formation to resorption. However, RANKL is expressed by a variety of cell types, and it is unclear which of them are essential sources for osteoclast formation. Here we have used a mouse strain in which RANKL can be conditionally deleted and a series of Cre-deleter strains to demonstrate that hypertrophic chondrocytes and osteocytes, both of which are embedded in matrix, are essential sources of the RANKL that controls mineralized cartilage resorption and bone remodeling, respectively. Moreover, osteocyte RANKL is responsible for the bone loss associated with unloading. Contrary to the current paradigm, RANKL produced by osteoblasts or their progenitors does not contribute to adult bone remodeling. These results suggest that the rate-limiting step of matrix resorption is controlled by cells embedded within the matrix itself.

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Figure 1: Deletion of RANKL in Prx1-Cre expressing cells causes osteopetrosis.
Figure 2: Deletion of RANKL in Osx1-Cre- and Ocn-Cre-expressing cells causes osteopetrosis.
Figure 3: Deletion of RANKL from Dmp1-Cre expressing cells reduces bone remodeling.
Figure 4: Osx1-Cre–mediated RANKL deletion in adult mice does not alter osteoclast number in cancellous bone.
Figure 5: Tail suspension of mice lacking RANKL in osteocytes.

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Acknowledgements

We thank P.E. Cazer, S.B. Berryhill, W. Webb, R. Shelton, A. Deloose, L.K. Climer, K. Vyas, L. Han, A.D. Warren, E.A. Hogan and J.J. Goellner for technical support and advice; and M. Almeida and H. Zhao for helpful discussions. We thank the following individuals for providing Cre-deleter strains: C.J. Tabin (Harvard Medical School), Prx1-Cre; H.M. Kronenberg (Harvard Medical School), Osx1-Cre; K. von der Mark (University of Erlangen-Nuremberg) and B. de Crombrugghe (M.D. Anderson Cancer Center), ColX-Cre; T.L. Clemens (Johns Hopkins University School of Medicine), Ocn-Cre; and J.Q. Feng (Baylor College of Dentistry) Dmp1-Cre. We thank P.D. Pajevic (Harvard Medical School) for the collagenase digestion protocol; D. Chen (University of Rochester School of Medicine) for the X-gal staining protocol; T. Bellido (Indiana University School of Medicine) and I. Aguirre (University of Florida) for advice on tail suspension; the staff of the UAMS Department of Laboratory Animal Medicine; and L. Suva and R. Skinner of the UAMS Skeletal Imaging Core. This work was supported by the following grants from the US National Institutes of Health: AR049794 (to C.A.O.) and AG13918 (to S.C.M.). Support was also provided by the Central Arkansas Veteran's Healthcare System (Merit Reviews to C.A.O., S.C.M., R.L.J. and R.S.W.), by the UAMS Translational Research Institute (1UL1RR029884) and by UAMS tobacco settlement funds.

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Contributions

J.X. carried out the conditional deletion breeding, analysis of gene expression, histomorphometry, immunochemistry and tail-suspension studies. M.O. carried out the R26R breeding and histological analysis of X-gal staining. C.A.O. designed experiments, created the RANKL-flox mice and prepared the first draft of the manuscript. R.L.J., R.S.W., S.C.M. and C.A.O. provided reagents, contributed methods, discussed results and revised the manuscript.

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Correspondence to Charles A O'Brien.

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Xiong, J., Onal, M., Jilka, R. et al. Matrix-embedded cells control osteoclast formation. Nat Med 17, 1235–1241 (2011). https://doi.org/10.1038/nm.2448

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