title: Specific inhibitors of vacuolar H(+)-ATPase trigger apoptotic cell death of osteoclasts.
abstract: Osteoclasts are multinucleated bone-resorbing cells that play a critical role in bone remodeling. Specific inhibitors of vacuolar H(+)-ATPase (V-ATPase), concanamycin A and bafilomycin A1, abolish bone resorption by osteoclasts. In this study, we examined whether these V-ATPase inhibitors trigger apoptotic cell death in osteoclasts, using murine osteoclast-like multinucleated cells (OCLs) formed in vitro. Acridine orange staining revealed that the treatment of OCLs with concanamycin A resulted in chromatin condensation and alterations in nuclear morphology within a few hours. The TdT-mediated dUTP-nick-end labeling (TUNEL) reaction confirmed the apoptotic features of OCLs treated with concanamycin A. The accelerated apoptotic cell death induced by concanamycin A occurred in OCLs treated with interleukin-1 alpha or macrophage colony-stimulating factor as well, which are known to elongate the survival time of osteoclasts. In contrast, these inhibitors did not induce cell death of osteoblastic cells isolated from mouse calvaria. These results suggest that functional impairment of V-ATPase triggers apoptotic cell death in osteoclasts.
authors.affiliation: Department of Oral Science, National Institute of Infectious Diseases, Tokyo, Japan.
authors.forename: N
authors.initials: N
authors.lastname: Okahashi
authors.forename: I
authors.initials: I
authors.lastname: Nakamura
authors.forename: E
authors.initials: E
authors.lastname: Jimi
authors.forename: M
authors.initials: M
authors.lastname: Koide
authors.forename: T
authors.initials: T
authors.lastname: Suda
authors.forename: T
authors.initials: T
authors.lastname: Nishihara
mesh_terms: D000818:Animals; D000900:Anti-Bacterial Agents / Q000494:pharmacology; D017209:Apoptosis / Q000187:drug effects*; D002478:Cells, Cultured; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D007375:Interleukin-1 / Q000494:pharmacology; D007700:Kinetics; D018942:Macrolides; D016173:Macrophage Colony-Stimulating Factor / Q000494:pharmacology; D051379:Mice; D010006:Osteoblasts / Q000166:cytology / Q000187:drug effects / Q000201:enzymology; D010010:Osteoclasts / Q000166:cytology / Q000187:drug effects* / Q000201:enzymology*; D006180:Proton-Translocating ATPases / Q000037:antagonists & inhibitors*; D025262:Vacuolar Proton-Translocating ATPases
chemical_list: D000900:Anti-Bacterial Agents; D004791:Enzyme Inhibitors; D007375:Interleukin-1; D018942:Macrolides; C036978:concanamycin A; D016173:Macrophage Colony-Stimulating Factor; C040929:bafilomycin A1; D025262:Vacuolar Proton-Translocating ATPases; D006180:Proton-Translocating ATPases