title: Inhibition of RANKL-dependent cellular fusion in pre-osteoclasts by amiloride and a NHE10-specific monoclonal antibody.
abstract: The functions of Na(+) /H(+) exchangers (NHEs) during osteoclastic differentiation were investigated using the NHE inhibitor amiloride and a monoclonal antibody (MAb). Compared with sRANKL-stimulated control cells, amiloride decreased the number of large TRAP-positive osteoclast cells (OCs) with ≥10 nuclei and increased the number of small TRAP-positive OCs with ≤10 nuclei during sRANKL-dependent osteoclastic differentiation of RAW264.7 cells. NHE10 mRNA expression and OC differentiation markers were increased by sRANKL stimulation in dose- and time-dependent manners. NHEs 1-9 mRNA expression was not increased by sRANKL stimulation. Similar to amiloride, a rat anti-mouse NHE10 MAb (clone 6B11) decreased the number of large TRAP-positive OCs, but increased the number of small TRAP-positive OCs. These findings suggested that inhibition of NHEs by amiloride or an anti-NHE10 MAb prevented sRANKL-promoted cellular fusion. The anti-NHE10 MAb has the potential for use as an effective inhibitor of bone resorption for targeted bone disease therapy.
authors.affiliation: Department of Oral Biology and Engineering, Integrated Health Sciences, Institute of Biomedical and Health Sciences, Hiroshima University, 1-2-3 Kasumi Minami-ku, Hiroshima, 734-8553, Japan.
authors.forename: Yuichi
authors.initials: Y
authors.lastname: Mine
authors.affiliation: Section of Fixed Prosthodontics, Division of Oral Rehabilitation, Faculty of Dental Science, Kyushu University, 3-1-1 Maidashi Higashi-ku, Fukuoka, 812-8582, Japan.
authors.forename: Takahiro
authors.initials: T
authors.lastname: Shuto
authors.affiliation: Department of Oral Biology and Engineering, Integrated Health Sciences, Institute of Biomedical and Health Sciences, Hiroshima University, 1-2-3 Kasumi Minami-ku, Hiroshima, 734-8553, Japan.
authors.forename: Hiroki
authors.initials: H
authors.lastname: Nikawa
authors.affiliation: Department of Immunology and Infectious Diseases, The Forsyth Institute, 245 1st St., Cambridge, MA, 02142, USA.|Department of Oral Medicine, Infection and Immunity, Harvard School of Dental Medicine, 188 Longwood Ave., Boston, MA, 02115,, USA.
authors.forename: Toshihisa
authors.initials: T
authors.lastname: Kawai
authors.affiliation: Hiroshima University Hospital, Dental Clinic, 1-1-2 Kagamiyama, Higashihiroshima, 739-0046, Japan.
authors.forename: Masaru
authors.initials: M
authors.lastname: Ohara
authors.affiliation: Department of Oral Biology and Engineering, Integrated Health Sciences, Institute of Biomedical and Health Sciences, Hiroshima University, 1-2-3 Kasumi Minami-ku, Hiroshima, 734-8553, Japan.
authors.forename: Kazuko
authors.initials: K
authors.lastname: Kawahara
authors.affiliation: Department of Immunology and Infectious Diseases, The Forsyth Institute, 245 1st St., Cambridge, MA, 02142, USA.|Department of Oral Medicine, Infection and Immunity, Harvard School of Dental Medicine, 188 Longwood Ave., Boston, MA, 02115,, USA.
authors.forename: Kouji
authors.initials: K
authors.lastname: Ohta
authors.affiliation: Department of Molecular Cell Biology and Oral Anatomy, Faculty of Dental Science, Kyushu University, 3-1-1 Maidashi Higashi-ku, Fukuoka, 812-8582, Japan.
authors.forename: Toshio
authors.initials: T
authors.lastname: Kukita
authors.affiliation: Section of Fixed Prosthodontics, Division of Oral Rehabilitation, Faculty of Dental Science, Kyushu University, 3-1-1 Maidashi Higashi-ku, Fukuoka, 812-8582, Japan.
authors.forename: Yoshihiro
authors.initials: Y
authors.lastname: Terada
authors.affiliation: Section of Fixed Prosthodontics, Division of Oral Rehabilitation, Faculty of Dental Science, Kyushu University, 3-1-1 Maidashi Higashi-ku, Fukuoka, 812-8582, Japan.
authors.forename: Seicho
authors.initials: S
authors.lastname: Makihira
mesh_terms: D000135:Acid Phosphatase / Q000378:metabolism; D000584:Amiloride / Q000494:pharmacology*; D000818:Animals; D000911:Antibodies, Monoclonal / Q000494:pharmacology*; D015415:Biomarkers / Q000378:metabolism; D002454:Cell Differentiation / Q000187:drug effects; D002459:Cell Fusion; D005786:Gene Expression Regulation / Q000187:drug effects; D007527:Isoenzymes / Q000378:metabolism; D008070:Lipopolysaccharides / Q000494:pharmacology; D051379:Mice; D008810:Mice, Inbred C57BL; D010010:Osteoclasts / Q000187:drug effects / Q000378:metabolism*; D010012:Osteogenesis / Q000187:drug effects; D053245:RANK Ligand / Q000494:pharmacology*; D000067996:RAW 264.7 Cells; D034622:RNA Interference / Q000187:drug effects; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D017208:Rats, Wistar; D060888:Real-Time Polymerase Chain Reaction; D017923:Sodium-Hydrogen Exchangers / Q000276:immunology*; D012995:Solubility; D000071681:Tartrate-Resistant Acid Phosphatase
chemical_list: D000911:Antibodies, Monoclonal; D015415:Biomarkers; D007527:Isoenzymes; D008070:Lipopolysaccharides; D053245:RANK Ligand; D012333:RNA, Messenger; D017923:Sodium-Hydrogen Exchangers; D000584:Amiloride; D000135:Acid Phosphatase; C000605511:Acp5 protein, mouse; D000071681:Tartrate-Resistant Acid Phosphatase
keywords: NHE10; RANKL; amiloride; cell-cell fusion; monoclonal antibody; osteoclast