title: Ca2+ or phorbol ester but not inflammatory stimuli elevate inducible nitric oxide synthase messenger ribonucleic acid and nitric oxide (NO) release in avian osteoclasts: autocrine NO mediates Ca2+-inhibited bone resorption.
abstract: Osteoclast bone resorption is essential for normal calcium homeostasis and is therefore tightly controlled by calciotropic hormones and local modulatory cytokines and factors. Among these is nitric oxide (NO), a multifunctional free radical that potently inhibits osteoclast bone resorption in vitro and in vivo. Previous findings led us to propose that NO might serve as an autocrine, as well as paracrine, regulator of osteoclast function. This premise was investigated using isolated bone-resorptive avian osteoclasts and focusing on the inducible isoform of NO synthase (iNOS) responsible for inflammatory stimulated high-level NO synthesis in other cells. Avian osteoclasts expressed both iNOS messenger RNA (mRNA) and protein. However, inflammatory cytokines that induce iNOS mRNA, protein, and NO in other cells did not do so in avian osteoclasts, consistent with the known role of inflammatory stimuli in promoting osteoclast resorption and localized bone loss. In searching for potential modulators of osteoclast iNOS, protein kinase C activation [by phorbol 12-myristate 13-acetate (PMA)] and intracellular Ca2+ rises (A23187) were each found to elevate osteoclast iNOS mRNA and protein levels, while increasing NO release and reducing osteoclast bone resorption. The iNOS selective inhibitor aminoguanidine suppressed stimulated osteoclast NO production elicited by either signal, but reversed only the resorption inhibition due to raised Ca2+. Thus, whereas additional inhibitory signals are presumably coproduced in osteoclasts treated with PMA, osteoclast iNOS-derived NO may act as an autocrine signal to mediate Ca2+-inhibited bone resorption. These findings document for the first time an iNOS whose mRNA levels are regulated by Ca2+ or PMA, but not inflammatory stimuli, and the autocrine production of NO as a Ca2+ sensing signal to suppress osteoclast bone resorption. The unusual regulation of osteoclast iNOS makes it a potentially attractive target for designing novel therapeutic agents to alleviate excessive bone loss.
authors.affiliation: Department of Biology, Washington University, St. Louis, Missouri 63130, USA.
authors.forename: T
authors.initials: T
authors.lastname: Sunyer
authors.forename: L
authors.initials: L
authors.lastname: Rothe
authors.forename: D
authors.initials: D
authors.lastname: Kirsch
authors.forename: X
authors.initials: X
authors.lastname: Jiang
authors.forename: F
authors.initials: F
authors.lastname: Anderson
authors.forename: P
authors.initials: P
authors.lastname: Osdoby
authors.forename: P
authors.initials: P
authors.lastname: Collin-Osdoby
mesh_terms: D000818:Animals; D001862:Bone Resorption; D001842:Bone and Bones; D000001:Calcimycin / Q000494:pharmacology; D002118:Calcium / Q000494:pharmacology*; D002478:Cells, Cultured; D002645:Chickens; D003470:Culture Media; D004789:Enzyme Activation; D015870:Gene Expression; D018836:Inflammation Mediators / Q000494:pharmacology; D007527:Isoenzymes / Q000235:genetics; D009569:Nitric Oxide / Q000378:metabolism*; D019001:Nitric Oxide Synthase / Q000235:genetics*; D009573:Nitrites / Q000378:metabolism; D010010:Osteoclasts / Q000378:metabolism*; D011493:Protein Kinase C / Q000378:metabolism; D012333:RNA, Messenger / Q000096:biosynthesis; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology*
chemical_list: D003470:Culture Media; D018836:Inflammation Mediators; D007527:Isoenzymes; D009573:Nitrites; D012333:RNA, Messenger; D009569:Nitric Oxide; D000001:Calcimycin; D019001:Nitric Oxide Synthase; D011493:Protein Kinase C; D013755:Tetradecanoylphorbol Acetate; D002118:Calcium