Cytoskeleton rearrangements during calcitonin-induced changes in osteoclast motility in vitro.
Cytoskeleton rearrangements during calcitonin-induced changes in osteoclast motility in vitro.
复制标题
降钙素诱导的体外破骨细胞运动变化过程中的细胞骨架重排。
DOI：
10.1016/8756-3282(85)90051-1
复制
发表时间：
1985
期刊：
影响因子：
4.1
通讯作者：
J. Heersche
中科院分区：
文献类型：
--
作者：
B. Warshafsky;Jane E. Aubin;J. Heersche
尽管对破骨细胞降解骨的了解相对较少，但体内和体外组织培养的观察表明破骨细胞运动是必需的。最近从大鼠股骨中分离出具有丰富运动活性和对骨调节激素降钙素有反应性的破骨细胞，并在培养物中短期保存。使用微管蛋白和肌球蛋白抗体的间接免疫荧光以及 F-肌动蛋白的 NBD-鬼笔酸染色，我们研究了未经处理或用降钙素处理的破骨细胞中的细胞骨架分布。未经处理和无反应的细胞在基质上很好地铺展，并在延时摄影中表现出惊人的运动性。在这些细胞中，可以看到精细的微丝网络，但看不到应力纤维。细胞外围和背侧细胞表面的褶皱均被 F-肌动蛋白强烈染色。相反，肌球蛋白染色以相对分散的点状图案出现，向细胞外围逐渐减弱，并且在褶皱边缘不存在。微管的标记模式与各种单核细胞中常见的模式类似，微管从核周区域向细胞外围辐射。在通过停止运动活动和收缩细胞质来响应降钙素的细胞中，微管没有发生可检测到的改变。然而，肌球蛋白和肌动蛋白标记都发生了巨大变化，回缩纤维对肌动蛋白进行了明亮的标记，但对肌球蛋白则没有标记。在细胞质回缩的早期，肌球蛋白在回缩纤维基部的细胞外围呈环状染色最强烈。这些数据表明，细胞骨架的特定成分会因刺激而改变，例如降钙素，肌动球蛋白相互作用可能参与细胞质回缩的早期阶段，并且这些成分可能在细胞响应该激素的分子机制中发挥积极作用。
Although relatively little is known about osteoclastic degradation of bone, observations both in vivo and in tissue culture in vitro suggest that osteoclast motility is required. Osteoclasts with abundant motile activity and responsiveness to a bone-regulating hormone, calcitonin, have recently been isolated from rat femur and maintained in culture for short periods of time. Using indirect immunofluorescence with antibodies to tubulin and myosin and NBD-phallacidin staining for F-actin, we have studied cytoskeleton distributions in such osteoclasts, either untreated or treated with calcitonin. Untreated and unresponsive cells were well spread on the substratum and displayed striking motility in time-lapse cinematography. In these cells a fine network of microfilaments but no stress fibers could be seen. Ruffles, both at the cell periphery and on the dorsal cell surface, stained intensely for F-actin. In contrast, myosin staining appeared in a relatively diffuse dotty pattern that diminished toward the cell periphery and was absent from the ruffled borders. Microtubules labeled in a pattern similar to that typically seen in a variety of mononuclear cells with microtubules radiating from the perinuclear region toward the cell periphery. In cells that responded to calcitonin by ceasing motile activity and retracting cytoplasm, microtubules were not detectably altered. Both myosin and actin labeling, however, changed dramatically, with retraction fibers labeled brightly for actin but not for myosin. Early in cytoplasmic retraction, myosin stained most intensely as a ring at the cell periphery at the base of retraction fibers. These data indicate that specific components of the cytoskeleton are altered in response to stimuli, such as calcitonin, that an actomyosin interaction may be involved in the early stages of cytoplasmic retraction, and that these components may play an active role in the molecular mechanisms by which cells respond to this hormone.
DOI：
10.1073/pnas.77.11.6624
发表时间：
1980-01-01
期刊：
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA-BIOLOGICAL SCIENCES
影响因子：
--
作者：
VERDERAME, M;ALCORTA, D;POLLACK, R
通讯作者：
POLLACK, R