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 Blood, 15 March 2006, Vol. 107, No. 6, pp. 2262-2270.
Prepublished online as a Blood First Edition Paper on November 22, 2005; DOI 10.1182/blood-2005-08-3365.

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Submitted August 18, 2005
Accepted November 7, 2005

Chemokine and chemokine receptor expression during colony stimulating factor-1-induced osteoclast differentiation in the toothless osteopetrotic rat: a key role for CCL9 (MIP-1{gamma}) in osteoclastogenesis in vivo and in vitro

Meiheng Yang, Genevieve Mailhot, Carole A MacKay, April Mason-Savas, Justin Aubin, and Paul R Odgren*

Department of Cell Biology, Univeristy of Massachusetts Medical School, Worcester, MA, USA

* Corresponding author; email: paul.odgren{at}umassmed.edu.

Osteoclasts differentiate from hematopoietic precursors under systemic and local controls. Chemokines and receptors direct leukocyte traffic throughout the body and may help regulate site-specific bone resorption. We investigated bone gene expression in vivo during rapid osteoclast differentiation induced by CSF-1 in Csf1-null toothless (tl/tl) rats. Long bone RNA from CSF-1-treated tl/tl rats was analyzed by high-density microarray over a time course. TRAP-positive osteoclasts appeared on day 2, peaked on day 4, and decreased slightly on day 6, as marrow space was expanding. TRAP and cathepsin K mRNA paralleled the cell counts. We examined all chemokine and receptor mRNAs on the arrays. CCL9 was strongly induced and peaked on day 2, as did its receptor, CCR1 and regulatory receptors c-Fms (CSF-1 receptor) and RANK. Other chemokines and receptors showed little or no significant changes. In situ hybridization and immunohistochemistry revealed CCL9 in small, immature osteoclasts on day 2 and in mature cells at later times. Anti-CCL9 antibody inhibited osteoclast differentiation in culture and significantly suppressed the osteoclast response in CSF-1 treated tl/tl rats. While various chemokines have been implicated in osteoclastogenesis in vitro, this first systematic analysis of chemokines and receptors during osteoclast differentiation in vivo highlights the key role of CCL9 in this process.


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