Research On The Role Of Tubulin In Osteoclast

With extended life expectancy and an increasing number of elderly population, as one of the senile degenerative disease of bone has become a serious social problem and has attracted much attention. Osteoporosis is a systemic metabolic bone disease, in recent years we using a variety of methods to research the osteoporosis etiology, and tries to elucidate the pathophysiological mechanisms. To maintain normal bone tissue based on bone resorption of osteoclast and bone formation of osteoblast in the dynamic balance, osteoclasts (OC) is considered to be the only cell which have the effect of bone absorption in the body, the activity of osteoclast change is closely related to the occurrence of a variety of metabolic bone disease, osteoclasts has long been a research focus in the bone metabolic disease.The cytoskeleton is a highly dynamic change of cell structure. At different times in the cell cycle, cell skeleton have completely different distribution state. Cytoskeleton plays a very important role in mechanical support and space location in cells, at the same time, the cytoskeleton is important eukaryotic cell structure and function of the organizer. Cytoskeleton is not only associated with the morphogenesis of cells, but also involved in almost all cells movement. In recent years studies have shown that cytoskeleton play an important role of in bone dissolution, it was found that microtubules and actin filaments have partly colocalization in cells, and Microtubules mediated podosome belt formation which can Influence osteoclast differentiation and maturation, bone dissolution have Close relationship with podosome belt formation. Microtubules in constant update changes, Microtubule dynamics may be associated with osteoclast activation which closely related to the formation of podosome belt. The purpose of our experiment is to study the microtubules mediated podosome formation mechanism, and find the feasible method intervention osteoclast dissolve bone.Our experiment established a stable transfection of osteoclasts precursor cells Raw264.7 cell line by liposome transfection method, and prove the stable transfection of Raw264.7 cells and normal Raw264.7 cells can be stable induced into osteoclasts. We used to live cell imaging techniques to observe protein EB1 in Raw264.7 cell and osteoclast intracellular distribution and movement, the results showed Raw264.7 cells and osteoclasts microtubules are constantly in the dynamic changes and the osteoclast microtubules dynamic rate slower than Raw264.7’. the part of a preliminary study on the experiment of microtubules in osteoclast function distribution and movement, This part of the experiment preliminary study of microtubules in osteoclast function of distribution and movement.Many studies have shown that the dynamic of the osteoclasts microtubules is associated with the formation of podosome belt. Osteoclast dynamic observed previous experiments with nocodazole and taxol to depolymerized microtubules and stabilizes microtubules respectively. However, the low concentration of nocodazole and taxol on osteoclast microtubules have not been reported. Low concentration of nocodazole and taxol has similarities to the role of microtubules, they can lower the dynamic of microtubules, the microtubules in the condition of no synthetic and no depolymerization. We use low concentration nocodazole(10nM) and Taxol(10nM) before osteoclast fusion to reduce osteoclasts precursor cells microtubule dynamics, experimental results show that low concentration of nocodazole and Taxol Influence the podosome belt formation in osteoclast fusion. Osteoclast formation with lower microtubule dynamics can reduce the number of osteoclast formation and lower tartrate resistant acid phosphatase activity by identifying tartrate resistant acid phosphatase (TRAP) staining and tartrate resistant acid phosphatase activity tests. Reduce osteoclasts microtubule activity is likely to be affected podosome precursor biosynthesis process, and form a disordered arrangement of podosome belt. These findings prompted us that osteoclast activity can be suppressed by the dose of a drug regulatory which affect osteoclasts microscopic protein, thereby reducing or delaying the occurrence of osteoporosis, there are important significance to bone metabolic diseases such as Osteoporosis treatment and prosthetic aseptic loosening after joint replacement.