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Adseverin knockdown inhibits osteoclastogenesis in RAW264.7 cells.

Qi W, Gao Y, Tian J, Jiang H - Int. J. Mol. Med. (2014)

Bottom Line: 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.

View Article: PubMed Central - PubMed

Affiliation: Department of Operative Dentistry and Endodontics, Guanghua School and Hospital of Stomatology, Guangdong Provincial Key Laboratory of Stomatology, Sun Yat‑sen University, Guangzhou, Guangdong 510055, P.R. China.

ABSTRACT
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.

No MeSH data available.


Adseverin (Ads) knockdown is associated with impaired RANK signaling. Signaling downstream of RANK was evaluated by nuclear factor-κB (NF-κB) and c-jun immunostaining on day 4 of osteoclastogenesis (OCG). NF-κB was activated in Luc-knockdown osteoclasts as shown by its exclusively nuclear distribution (arrows). The majority Ads-knockdown macrophages did not exhibit active NF-κB, as shown by its cytoplasmic distribution. Luc knockdown and Ads knockdown demonstrated activated nuclear c-jun, a transcription factor involved in RANK survival signaling in osteoclasts. RANK, receptor activator of nuclear factor-κB.
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f6-ijmm-34-06-1483: Adseverin (Ads) knockdown is associated with impaired RANK signaling. Signaling downstream of RANK was evaluated by nuclear factor-κB (NF-κB) and c-jun immunostaining on day 4 of osteoclastogenesis (OCG). NF-κB was activated in Luc-knockdown osteoclasts as shown by its exclusively nuclear distribution (arrows). The majority Ads-knockdown macrophages did not exhibit active NF-κB, as shown by its cytoplasmic distribution. Luc knockdown and Ads knockdown demonstrated activated nuclear c-jun, a transcription factor involved in RANK survival signaling in osteoclasts. RANK, receptor activator of nuclear factor-κB.

Mentions: RANK downstream signaling was evaluated using NF-κB immunostaining on day 4 in Ads-knockdown and Luc-knockdown cells. NF-κB was activated in Luc-knockdown osteoclasts only, as shown by its exclusive nuclear distribution (arrows). Ads-knockdown monocytes did not exhibit active NF-κB as shown by its cytoplasmic distribution. However, monocytes and osteoclasts in the two groups demonstrated activated nuclear c-jun, a transcription factor involved in RANK signaling associated with apoptosis (Fig. 6).


Adseverin knockdown inhibits osteoclastogenesis in RAW264.7 cells.

Qi W, Gao Y, Tian J, Jiang H - Int. J. Mol. Med. (2014)

Adseverin (Ads) knockdown is associated with impaired RANK signaling. Signaling downstream of RANK was evaluated by nuclear factor-κB (NF-κB) and c-jun immunostaining on day 4 of osteoclastogenesis (OCG). NF-κB was activated in Luc-knockdown osteoclasts as shown by its exclusively nuclear distribution (arrows). The majority Ads-knockdown macrophages did not exhibit active NF-κB, as shown by its cytoplasmic distribution. Luc knockdown and Ads knockdown demonstrated activated nuclear c-jun, a transcription factor involved in RANK survival signaling in osteoclasts. RANK, receptor activator of nuclear factor-κB.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4214352&req=5

f6-ijmm-34-06-1483: Adseverin (Ads) knockdown is associated with impaired RANK signaling. Signaling downstream of RANK was evaluated by nuclear factor-κB (NF-κB) and c-jun immunostaining on day 4 of osteoclastogenesis (OCG). NF-κB was activated in Luc-knockdown osteoclasts as shown by its exclusively nuclear distribution (arrows). The majority Ads-knockdown macrophages did not exhibit active NF-κB, as shown by its cytoplasmic distribution. Luc knockdown and Ads knockdown demonstrated activated nuclear c-jun, a transcription factor involved in RANK survival signaling in osteoclasts. RANK, receptor activator of nuclear factor-κB.
Mentions: RANK downstream signaling was evaluated using NF-κB immunostaining on day 4 in Ads-knockdown and Luc-knockdown cells. NF-κB was activated in Luc-knockdown osteoclasts only, as shown by its exclusive nuclear distribution (arrows). Ads-knockdown monocytes did not exhibit active NF-κB as shown by its cytoplasmic distribution. However, monocytes and osteoclasts in the two groups demonstrated activated nuclear c-jun, a transcription factor involved in RANK signaling associated with apoptosis (Fig. 6).

Bottom Line: 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.

View Article: PubMed Central - PubMed

Affiliation: Department of Operative Dentistry and Endodontics, Guanghua School and Hospital of Stomatology, Guangdong Provincial Key Laboratory of Stomatology, Sun Yat‑sen University, Guangzhou, Guangdong 510055, P.R. China.

ABSTRACT
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.

No MeSH data available.