Adseverin knockdown inhibits osteoclastogenesis in RAW264.7 cells

Osteoclastogenesis is a complex process that is highly dependent on the dynamic regulation of the actin cytoskeleton. Adseverin (Ads), a member of the gelsolin superfamily of actin-binding proteins, regulates actin remodeling by severing and capping actin filaments. The objective of the present study was to characterize the role of Ads during osteoclastogenesis by assessing Ads expression and using a knockdown strategy. Immunoblot analyses were used to examine Ads expression during osteoclastogenesis. A stable Ads knockdown macrophage cell line was generated using a retroviral shRNA construct. Osteoclast differentiation was morphologically examined via cell staining with osteoclast specific markers and light microscopy. The results showed that Ads expression was significantly increased in response to receptor activator of nuclear factor-κB ligand during osteoclastogenesis, and Ads was highly expressed in mature osteoclasts. Ads-knockdown macrophages showed major osteoclastogenesis defects, most likely caused by a pre-osteoclast fusion defect. These results indicate that Ads deficiency in monocytes inhibits osteoclastogenesis. Thus, in future studies it could be noteworthy to investigate the function of Ads in bone marrow monocytes during osteoclastogenesis.