Premium
RGS12 Is Essential for RANKL‐Evoked Signaling for Terminal Differentiation of Osteoclasts In Vitro
Author(s) -
Yang Shuying,
Li YiPing
Publication year - 2007
Publication title -
journal of bone and mineral research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.882
H-Index - 241
eISSN - 1523-4681
pISSN - 0884-0431
DOI - 10.1359/jbmr.061007
Subject(s) - rankl , osteoclast , microbiology and biotechnology , rna interference , cellular differentiation , gene silencing , signal transduction , biology , chemistry , in vitro , gene , rna , biochemistry , activator (genetics)
How RANKL evokes [Ca 2+ ] i oscillations and leads to osteoclast differentiation is unclear. We identified a new signaling protein, RGS12, and found that RGS12 is essential for [Ca 2+ ] i oscillations and osteoclast differentiation induced by RANKL. RGS12 may play a critical role in the RANKL‐evoked PLCγ–calcium channels–[Ca 2+ ] i oscillation–NFAT2 pathway. Introduction : RANKL‐induced [Ca 2+ ] i oscillations play a switch‐on role in NFAT2 expression and osteoclast differentiation. However, RANKL evokes [Ca 2+ ] i oscillations and leads to osteoclast differentiation by an unknown mechanism. In this study, we identified a new RANKL‐induced signaling protein, regulator of G signaling protein 12 (RGS12), and investigated its effect on osteoclast differentiation in vitro. Materials and Methods : We used a genome‐wide screening approach to identify genes that are specifically or prominently expressed in osteoclasts. To study the role of the RGS12 in osteoclast differentiation, we used vector and lentivirus‐based RNAi gene silencing technology to silence the RGS12 gene in the monocyte progenitor cell lines and primary bone marrow–derived monocytes (BMMs). The interaction between RGS12 and N‐type calcium channels was elucidated using co‐immunoprecipitation and immunoblotting. Results : We found that RGS12 was prominently expressed in osteoclast‐like cells (OLCs) induced by RANKL. This result was further confirmed at both the mRNA and protein level in human osteoclasts and mouse OLCs. Silence of RGS12 expression using vector and lentivirus based RNA interference (RNAi) impaired phosphorylation of phospholipase C (PLC)γ and blocked [Ca 2+ ] i oscillations, NFAT2 expression, and osteoclast differentiation in RANKL‐induced RAW264.7 cells and BMMs. We further found that N‐type calcium channels were expressed in OLCs after RANKL stimulation and that RGS12 directly interacted with the N‐type calcium channels. Conclusions : These results reveal that RGS12 is essential for the terminal differentiation of osteoclasts induced by RANKL. It is possible that RGS12 regulates osteoclast differentiation through a PLC γ–calcium channel–[Ca 2+ ] i oscillation–NFAT2 pathway.