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TRPV-5 mediates a receptor activator of NF-kappaB (RANK) ligand-induced increase in cytosolic Ca2+ in human osteoclasts and down-regulates bone resorption Chamoux E; Bisson M; Payet MD; Roux SJ Biol Chem 2010[Aug]; 285 (33): 25354-62Most of the signaling effectors located downstream of receptor activator of NF-kappaB (RANK) activation are calcium-sensitive. However, the early signaling events that lead to the mobilization of intracellular calcium in human osteoclasts are still poorly understood. The Ca(2+)-sensitive fluorescent probe Fura2 was used to detect changes in the intracellular concentration of Ca(2+) ([Ca(2+)](i)) in a model of human osteoclasts. Stimulating these cells with receptor activator of NF-kappaB ligand (RANKL) induced a rapid and significant increase in [Ca(2+)](i). Adding extracellular Ca(2+) chelators, depleting intracellular stores, and the use of a phospholipase C inhibitor all indicated that the Ca(2+) was of extracellular origin, suggesting the involvement of a Ca(2+) channel. We showed that none of the classical Ca(2+) channels (L-, T-, or R-type) were involved in the RANKL-induced Ca(2+) spike. However, the effect of high doses of Gd(3+) did suggest that TRP family channels were present in human osteoclasts. The TRPV-5 channel was expressed in osteoclasts and was mainly located in the cellular area in contact with the bone surface. Furthermore, the RNA inactivation of TRPV-5 channel completely inhibited the RANKL-induced increase in [Ca(2+)](i), which was accompanied in the long term by marked activation of bone resorption. Overall, our results show that RANKL induced a significant increase in [Ca(2+)](i) of extracellular origin, probably as a result of the opening of TRPV-5 calcium channels on the surface of human osteoclasts. Our findings suggest that TRPV-5 contributes to maintaining the homeostasis of the human skeleton via a negative feedback loop in RANKL-induced bone resorption.|Animals[MESH]|Blotting, Western[MESH]|Bone Resorption/genetics/*metabolism[MESH]|Calcium/*metabolism[MESH]|Cattle[MESH]|Cells, Cultured[MESH]|Cytosol/drug effects/*metabolism[MESH]|Female[MESH]|Fluorescent Antibody Technique[MESH]|Humans[MESH]|Microscopy, Confocal[MESH]|Osteoclasts/*drug effects/*metabolism[MESH]|Polymerase Chain Reaction[MESH]|RANK Ligand/*pharmacology[MESH]|Signal Transduction/drug effects[MESH]|TRPV Cation Channels/genetics/*metabolism[MESH] |
