Unknown,Transcriptomics,Genomics,Proteomics

Dataset Information

Gene expression analysis of transcriptome changes regulated by RANKL-RANK signaling in macrophage cell line RAW 264.7

ABSTRACT: Bone remodeling relies on the precise developmental control of osteoclasts that resorb bone and osteoblasts that lay down fresh bone matrix. The hematopoietically derived osteoclast follow a tigthly controlled genetic program where specific genes are up-regulated to aid bone resorption.Some of them for instance control membrane polarization, others transporters and pumps to move the acid and proteases intot he resorption pit. We used microarrays to detail the global programme of gene expression underlying differentiation of osteoclasts and identified distinct classes of up-regulated genes during this process. RAW cells were selected at different time periods of culture during the osteoclast differentiation process for RNA extraction and hybridization on Affymetrix microarrays. We sought to obtain more homogenous temporal resolution of expression profiles by using a cell line RAW 264.7 and including data of control cells that had been cultured for similar time periods without RANKL.

ORGANISM(S): Mus musculus

SUBMITTER: Parul Sharma

PROVIDER: E-GEOD-21639 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Publications

RANKL-RANK signaling regulates expression of xenotropic and polytropic virus receptor (XPR1) in osteoclasts.

Sharma Parul P Patntirapong Somying S Hann Steven S Hauschka Peter V PV

Biochemical and biophysical research communications 20100713 2

Formation of multinucleated bone-resorbing osteoclasts results from activation of the receptor activated NF-kappaB ligand (RANKL)-receptor activated NF-kappaB (RANK) signaling pathway in primary bone marrow macrophages and a macrophage cell line (RAW 264.7). Osteoclasts, through bone remodeling, are key participants in the homeostatic regulation of calcium and phosphate levels within the body. Microarray analysis using Gene Expression Dynamic Inspector (GEDI) clustering software indicated that o ...[more]

PMID: 20633538

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Project description:Bone remodeling is characterized by the sequential, local tethering of osteoclasts and osteoblasts, and is key to the maintenance of bone integrity. While bone matrix-mobilized growth factors, such as TGF-β, are proposed to regulate remodeling, no in vivo evidence exists that an osteoclast-produced molecule is the enigmatic coupling factor. We have identified Cthrc1, a protein secreted by mature bone-resorbing osteoclasts, that targets stromal cells so as to stimulate osteogenesis. The expression of Cthrc1 is robustly induced when mature osteoclasts are placed on dentin or hydroxyapatite, and also by increasing extracellular calcium. Cthrc1 expression in bone increases in a high turnover state, such as that which is induced by RANKL injections in vivo, whereas it decreases with aging or following alendronate treatment, conditions associated with suppressed bone turnover. The targeted deletion of the Cthrc1 gene eliminates Cthrc1 expression in bone, whereas its deficiency in osteoblasts does not exert any significant effect. Osteoclast-specific deletion of the Cthrc1 gene results in osteopenia due to reduced bone formation: it also impairs the coupling process following resorption induced by RANKL injections, with a resultant impairment of bone mass recovery. Thus, Cthrc1 is an osteoclast-secreted “coupling factor” that regulates bone remodeling and hence, skeletal integrity. Total bone marrow cells were prepared from the femurs and tibias of 8-10-week-old C57BL/6 mice and cultured in the presence of M-CSF (100ng/ml) for 3 days as described previously (Takeshita et al., 2000 JBMR 15:1477-1488). Cells were harvested with 0.02% EDTA/PBS and used as bone marrow macrophages (BMMs). These BMMs were cultured in the presence of M-CSF (100 ng/ml) and RANKL (100ng/ml) for 2 days. TRAP positive mononuclear cells were harvested and used as pre-osteoclasts (pOC). These pOC cells were further cultured in the presence of M-CSF and RANKL for 2 days in normal plastic plate or on dentin slices. After 2 days, multinucleated TRAP positive mature osteoclasts were generated as mature osteoclasts on plate (mOCp) and mature resorbing osteoclasts on dentin (mOCd), respectively. RNAs were extracted from four different stages of osteoclast lineage cells; BMMs, pOC, mOCp and mOCd, and used for microarray analysis.

Dataset Information

Gene expression analysis of transcriptome changes regulated by RANKL-RANK signaling in macrophage cell line RAW 264.7

Publications

RANKL-RANK signaling regulates expression of xenotropic and polytropic virus receptor (XPR1) in osteoclasts.

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