Effects of Inositol 1,4,5-triphosphate on Osteoclast Differentiation in RANKL-induced Osteoclastogenesis

• Published in: The Korean journal of physiology & pharmacology 2012, 16(1), , pp.31-36
• Main Authors: Son, Aran, Kim, Min Seuk, Jo, Hae, Byun, Hae Mi, Shin, Dong Min
• Format: Article
• Language: English
• Subjects: Original; 약리학
• ISSN: 1226-4512; 2093-3827
• DOI: 10.4196/kjpp.2012.16.1.31

The receptor activator of NF-κB ligand (RANKL) signal is an activator of tumor necrosis factor receptor-associated factor 6 (TRAF6), which leads to the activation of NF-κB and other signal transduction pathways essential for osteoclastogenesis, such as Ca 2+ signaling. However, the intracellular levels of inositol 1,4,5-trisphosphate (IP 3 ) and IP 3 -mediated cellular function of RANKL during osteoclastogenesis are not known. In the present study, we determined the levels of IP 3 and evaluated IP 3 -mediated osteoclast differentiation and osteoclast activity by RANKL treatment of mouse leukemic macrophage cells (RAW 264.7) and mouse bone marrow-derived monocyte/macrophage precursor cells (BMMs). During osteoclastogenesis, the expression levels of Ca 2+ signaling proteins such as IP 3 receptors (IP 3 Rs), plasma membrane Ca 2+ ATPase, and sarco/endoplasmic reticulum Ca 2+ ATPase type2 did not change by RANKL treatment for up to 6 days in both cell types. At 24 h after RANKL treatment, a higher steady-state level of IP 3 was observed in RAW264.7 cells transfected with green fluorescent protein (GFP)-tagged pleckstrin homology (PH) domains of phospholipase C (PLC) δ, a probe specifically detecting intracellular IP 3 levels. In BMMs, the inhibition of PLC with U73122 [a specific inhibitor of phospholipase C (PLC)] and of IP 3 Rs with 2-aminoethoxydiphenyl borate (2APB; a non-specific inhibitor of IP 3 Rs) inhibited the generation of RANKL-induced multinucleated cells and decreased the bone-resorption rate in dentin slice, respectively. These results suggest that intracellular IP 3 levels and the IP 3 -mediated signaling pathway play an important role in RANKL-induced osteoclastogenesis.