Inhibition of RANKL-dependent cellular fusion in pre-osteoclasts by amiloride and a NHE10-specific monoclonal antibody

Yuichi Mine, Takahiro Shuto, Hiroki Nikawa, Toshihisa Kawai, Masaru Ohara, Kazuko Kawahara, Kouji Ohta, Toshio Kukita, Yoshihiro Terada, Seicho Makihira

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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.

Original languageEnglish
Pages (from-to)696-709
Number of pages14
JournalCell Biology International
Volume39
Issue number6
DOIs
Publication statusPublished - Jun 1 2015

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Amiloride
Osteoclasts
Sodium-Hydrogen Antiporter
Monoclonal Antibodies
Cell Differentiation
Bone Density Conservation Agents
Messenger RNA
Bone Diseases
Differentiation Antigens
Clone Cells
Cell Count

All Science Journal Classification (ASJC) codes

  • Cell Biology

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Inhibition of RANKL-dependent cellular fusion in pre-osteoclasts by amiloride and a NHE10-specific monoclonal antibody. / Mine, Yuichi; Shuto, Takahiro; Nikawa, Hiroki; Kawai, Toshihisa; Ohara, Masaru; Kawahara, Kazuko; Ohta, Kouji; Kukita, Toshio; Terada, Yoshihiro; Makihira, Seicho.

In: Cell Biology International, Vol. 39, No. 6, 01.06.2015, p. 696-709.

Research output: Contribution to journalArticle

Mine, Y, Shuto, T, Nikawa, H, Kawai, T, Ohara, M, Kawahara, K, Ohta, K, Kukita, T, Terada, Y & Makihira, S 2015, 'Inhibition of RANKL-dependent cellular fusion in pre-osteoclasts by amiloride and a NHE10-specific monoclonal antibody', Cell Biology International, vol. 39, no. 6, pp. 696-709. https://doi.org/10.1002/cbin.10447
Mine, Yuichi ; Shuto, Takahiro ; Nikawa, Hiroki ; Kawai, Toshihisa ; Ohara, Masaru ; Kawahara, Kazuko ; Ohta, Kouji ; Kukita, Toshio ; Terada, Yoshihiro ; Makihira, Seicho. / Inhibition of RANKL-dependent cellular fusion in pre-osteoclasts by amiloride and a NHE10-specific monoclonal antibody. In: Cell Biology International. 2015 ; Vol. 39, No. 6. pp. 696-709.
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