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A RANKL-based Osteoclast Culture Assay of Mouse Bone Marrow to Investigate the Role of mTORC1 in Osteoclast Formation
Published on: March 15, 2018
Inhibition of microtubule dynamics affects podosome belt formation during osteoclast induction
Yunfan Ti1, Lingjun Zhou, Rui Wang
1Department of Orthopedics of Jinling Hospital, School of Medicine, Nanjing University, 305 Zhongshan East Rd, Nanjing, 210002, China.
Abstract:
Osteoclast is the only cell that can degrade bone tissue in vivo. Recent studies have shown the important role of cytoskeleton dynamics in osteolysis and the formation of podosome belt in osteoclasts. This process is regulated by the dynamic microtubule (MT) network. We treated osteoclast precursor cells Raw264.7 with low concentration of nocodazole (10 nM) and antineoplastic drug taxol (10 nM) to block MT turnover, and used end binding protein 1 fused to GFP to track the movement of microtubules in induced osteoclasts. We show that low concentrations of nocodazole and taxol interfere with the formation of podosome belt, and reduce TRAP activity of induced osteoclasts. These results suggest that the effect of taxol on MT dynamics may be used clinically to reduce osteoclast activity and potentially prevent development of osteoporosis and other metabolic bone diseases.
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