論文 - 詳細
| RRC ID | 18749 |
|---|---|
| 著者 | Sakai H, Moriura Y, Notomi T, Kawawaki J, Ohnishi K, Kuno M. |
| タイトル | Phospholipase C-dependent Ca2+-sensing pathways leading to endocytosis and inhibition of the plasma membrane vacuolar H+-ATPase in osteoclasts. |
| ジャーナル | Am J Physiol Cell Physiol |
| Abstract |
In osteoclasts, elevation of extracellular Ca2+ is an endogenous signal that inhibits bone resorption. We recently found that an elevation of extracellular Ca2+ decreased proton extrusion through the plasma membrane vacuolar H+-ATPase (V-ATPase) rapidly. In this study we investigated mechanisms underlying this early Ca2+-sensing response, particularly in reference to the activity of the plasma membrane V-ATPase and to membrane retrieval. Whole cell clamp recordings allowed us to measure the V-ATPase currents and the cell capacitance (C(m)) simultaneously. C(m) is a measure of cell surface. Extracellular Ca2+ (2.5-40 mM) decreased C(m) and the V-ATPase current simultaneously. The decreased C(m), together with the enhanced uptake of a lipophilic dye (FM1-43), indicated that Ca2+ facilitated endocytosis. The endocytosis was blocked by dynamin inhibitors (dynasore and dynamin-inhibitory peptide), by small interfering RNA (siRNA) targeting for dynanmin-2 and also by bafilomycin A(1), a blocker of V-ATPases. The extracellular Ca2+-induced endocytosis and inhibition of the V-ATPase current were diminished by a phospholipase C inhibitor (U73122) and siRNA targeting for phospholipase C gamma2 subunit. Holding the cytosolic Ca2+ at either high (0.5-5 microM) or low levels or inhibiting calmodulin by an inhibitor (W7) or an antibody (anti-CaM) decreased the stimulated endocytosis and the inhibition of the V-ATPase current. These data suggest that extracellular Ca2+ facilitated dynamin- and V-ATPase-dependent endocytosis in association with an inhibition of the plasma membrane V-ATPase. Phospholipase C, cytosolic Ca2+, and calmodulin were involved in the signaling pathways. Membrane retrieval and the plasma membrane V-ATPase activity may cooperate during the early phase of Ca2+-sensing response in osteoclasts. |
| 巻・号 | 299(3) |
| ページ | C570-8 |
| 公開日 | 2010-9-1 |
| DOI | 10.1152/ajpcell.00486.2009 |
| PII | ajpcell.00486.2009 |
| PMID | 20592242 |
| MeSH | Animals Calcium / metabolism* Calmodulin / metabolism Cell Line Cell Membrane / metabolism* Cytosol / metabolism Dynamins / metabolism Electric Capacitance Endocytosis* Extracellular Space / metabolism Mice Osteoclasts / metabolism* Patch-Clamp Techniques Signal Transduction Type C Phospholipases / physiology* Vacuolar Proton-Translocating ATPases / antagonists & inhibitors* Vacuolar Proton-Translocating ATPases / physiology |
| IF | 3.485 |
| 引用数 | 21 |
| WOS 分野 | PHYSIOLOGY CELL BIOLOGY |
| オルトメトリクス指標 |
オルトメトリクス指標項目
|
| 各媒体での言及数の合計 | 0 |
| リソース情報 | |
| ヒト・動物細胞 | RAW 264(RCB0535) |