Single cell mechanotransduction and its modulation analyzed by atomic force microscope indentation
Single cell mechanotransduction and its modulation analyzed by atomic force microscope indentation
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DOI：
10.1016/s0006-3495(02)75638-5
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发表时间：
2002-06-01
影响因子：
3.4
通讯作者：
Horton, MA
中科院分区：
文献类型：
--
作者：
Charras, GT;Horton, MA
骨骼适应其机械使用，尽管在细胞水平上，产生的应变的分布和大小及其检测还不清楚。使用原子力显微镜（AFM）作为微压痕仪，研究了细胞响应的应变的大小和性质。与该装置连接的共聚焦显微镜能够分析细胞反应。确定了两种不同的细胞反应途径：一种是接触后，依赖于拉伸激活离子通道的激活;第二种是应力松弛后，需要一个完整的微管细胞骨架。通过选择性地破坏细胞骨架成分来调节细胞反应，这些细胞骨架成分被认为参与机械刺激的转导。F-肌动蛋白细胞骨架是不需要的机械应变的反应，而微管和波形蛋白网络。降低膜张力或其传递的处理选择性地减少了接触反应。细胞骨架的免疫染色用于解释细胞骨架破坏研究的结果。我们提供了引起细胞内钙反应所需的细胞应变幅度的估计，并提出了一个模型，将单细胞反应与整个骨适应联系起来。这项技术可能有助于理解其他器官对机械使用的适应。
The skeleton adapts to its mechanical usage, although at the cellular level, the distribution and magnitude of strains generated and their detection are ill-understood. The magnitude and nature of the strains to which cells respond were investigated using an atomic force microscope (AFM) as a microindentor. A confocal microscope linked to the setup enabled analysis of cellular responses. Two different cell response pathways were identified: one, consequent upon contact, depended on activation of stretch-activated ion channels; the second, following stress relaxation, required an intact microtubular cytoskeleton. The cellular responses could be modulated by selectively disrupting cytoskeletal components thought to be involved in the transduction of mechanical stimuli. The F-actin cytoskeleton was not required for responses to mechanical strain, whereas the microtubular and vimentin networks were. Treatments that reduced membrane tension, or its transmission, selectively reduced contact reactions. Immunostaining of the cell cytoskeleton was used to interpret the results of the cytoskeletal disruption studies. We provide an estimate of the cellular strain magnitude needed to elicit intracellular calcium responses and propose a model that links single cell responses to whole bone adaptation. This technique may help to understand adaptation to mechanical usage in other organs.