The role of cytoskeleton on the transaction of mechanical stress to biological response in osteocytes -Transfection of GFP gene to cytoskeleton

细胞骨架在骨细胞机械应力与生物反应的转换中的作用-GFP基因转染到细胞骨架

• 批准号: 12671998
• 负责人: KAMIOKA Hiroshi
• 金额: $ 0.38万
• 依托单位: OKAYAMA UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2001
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-12671998/
• 关键词: mechanical stress; osteocytes; osteoblasts; cytoskeleton; connective tissue growth factor; fluid shear stress; connective tissue growth factor

Morphological changes of cytoskeleton in osteoblasts and osteocytes were investigated after subjection to fluid shear stress. It was observed that osteoblastic cell line MC3T3-E1 react to fluid shear stress and changes its cytoskeleton (especially, actin) after application of fluid shear stress of 12 dynes/cm2 for 1 hour. Hence, other cytoskeletal protein, such as vimentin and tubulin did not - change their morphology after adaptation of fluid shear stress. When the cells were microinjected with fluoresncence-labeled vinculin and subjected to fluid shear stress, it was confirmed that the assembly of vinculin was performed within 10 minutes after application of fluid shear stress. So, it was suggested that morphological changes after fluid shear stress happens very rapidly. However, when osteocytes were subjected to fluid shear stress, the cytoskeletal changes was not observed. Based on these results, the cytoskeleton in osteocytes was very stable comparing to osteoblasts. Next, we focused on the stability of osteocyte morphology and observed the three dimensional distribution of osteocyte processes in bone using confocal microscopy and differential interference contrast microscopy.Furthermore, we found that connective tissue growth factor is a candidate of marker of cytoskeletal changes in bone cells.

研究了受到流体剪切应力后成骨细胞和骨细胞的细胞骨架的形态变化。观察到成骨细胞系MC3T3-E1在施加12达因/cm2的流体剪切应力1小时后对流体剪切应力做出反应并改变其细胞骨架（特别是肌动蛋白）。因此，其他细胞骨架蛋白，例如波形蛋白和微管蛋白，在适应流体剪切应力后不会改变其形态。当向细胞显微注射荧光标记的纽蛋白并施加流体剪切应力时，证实了在施加流体剪切应力后10分钟内进行了纽蛋白的组装。因此，有人认为流体剪切应力后的形态变化发生得非常迅速。然而，当骨细胞受到流体剪切应力时，没有观察到细胞骨架的变化。基于这些结果，与成骨细胞相比，骨细胞的细胞骨架非常稳定。接下来，我们关注骨细胞形态的稳定性，并利用共聚焦显微镜和微分干涉显微镜观察骨中骨细胞突起的三维分布。此外，我们发现结缔组织生长因子是骨细胞中细胞骨架变化的候选标志物。

项目成果

T. YAMASHIRO, K. FUJIYAMA, Y FUJIYOSHl, N. INAGAKI, T. TAKANO-YAMAMOTO: "INFERIOR ALVEOLAR NERVE TRANSFECTION INHIBITS INCREASE IN OSTEOCLAST APPEARANCE"BONE. 26. 663-669 (2001)

T. YAMASHIRO、K. FUJIYAMA、Y FUJIYOSHl、N. INAGAKI、T. TAKANO-YAMAMOTO：“下牙槽神经转染抑制破骨细胞出现的增加”骨。

Tusboi, Y.: "Mitogenic effect of neurotrophins of periodontal ligament cell line"Journal of Dental Research. 80・3. 181-186 (2001)

Tusboi，Y.：“牙周膜细胞系的神经营养素的促有丝分裂作用”牙科研究杂志80・3（2001）。

H. KAMIOKA, T. HONJO, T. TAKANO-YAMAMOTO: "A THREE DIMENSIONAL DISTRIBUTION OF OSTEOCYTE PROCESSES REVEALED BY THE COMBINATION OF CONFOCAL LASER SCANNING MICROSCOPY AND DIFFERENTIAL INTERFERENCE CONTRAST MICROS COPY."BONE. 28. 145-149 (2001)

H. KAMIOKA、T. HONJO、T. TAKANO-YAMAMOTO：“通过共焦激光扫描显微镜和微分干涉对比显微镜相结合揭示骨细胞过程的三维分布。”骨头。

T. YAMASHlRO, H. KABUTO, T. FUKUNAGA, N. OGAWA, T. TAKANO-YAMAMOTO: "MEDULLARY MONOAMINE LEVELS DURING EXPERIMENTAL TOOTH MOVEMENT."BRAIN RESEARCH. 878. 199-203 (2000)

T. YAMASHIRO、H. KABUTO、T. FUKUNAGA、N. OGAWA、T. TAKANO-YAMAMOTO：“实验性牙齿移动期间的髓质单胺水平。”大脑研究。

Yamashiro,T.: "Inferior alveolar nerve transection inhibits increase in osteoclast appearance during experimental tooth movement"Bone. 26・6. 663-669 (2000)

Yamashiro，T.：“下牙槽神经横断抑制实验性牙齿运动期间破骨细胞出现的增加”Bone 663-669（2000）。