Patterning, prestress, and peeling dynamics of myocytes
Patterning, prestress, and peeling dynamics of myocytes
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DOI：
10.1016/s0006-3495(04)74195-8
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发表时间：
2004-02-01
影响因子：
3.4
通讯作者：
Discher, DE
中科院分区：
文献类型：
--
作者：
Griffin, MA;Engler, AJ;Discher, DE
作为典型的锚定依赖性细胞，肌细胞必须平衡收缩性和足够的粘附性。从成肌细胞中分离出的骨骼肌管在微图纹玻璃上生长，在机械分离肌管一端后，肌管表现出自发剥离。这些结果表明在细胞中产生了一种预应力。为了评估这种预应力并研究单个肌细胞的动态粘附强度，然后使用大口径微管吸入液体的剪切应力强制剥离肌管。细胞从表面剥离的速度（V-peel）被测量为施加张力（T-peel）的连续增加函数，其范围从；0 ~ 50 nN/mum。对于每个细胞，剥离被证明是高度异质性的，V-peel在0 mum/s（类似于80%的时间）和10 mum/s之间波动。对表达GFP-paxillin的平滑肌细胞的平行研究也显示出不连续的剥离，在强附着部位的局部粘连断裂（当使用小孔移液管进行压力剥离时）。本文描述的剥离方法有助于深入了解收缩-粘附平衡，并可通过物理检测和使用GFP标记来研究应力粘附的实时动态；这些方法在比较正常和营养不良的肌肉细胞方面应该是有用的。
As typical anchorage-dependent cells myocytes must balance contractility against adequate adhesion. Skeletal myotubes grown as isolated strips from myoblasts on micropatterned glass exhibited spontaneous peeling after one end of the myotube was mechanically detached. Such results indicate the development of a prestress in the cells. To assess this prestress and study the dynamic adhesion strength of single myocytes, the shear stress of fluid aspirated into a large-bore micropipette was then used to forcibly peel myotubes. The velocity at which cells peeled from the surface, V-peel, was measured as a continuously increasing function of the imposed tension, T-peel, which ranges from; 0 to 50 nN/mum. For each cell, peeling proved highly heterogeneous, with V-peel fluctuating between 0 mum/s (similar to80% of time) and similar to10 mum/s. Parallel studies of smooth muscle cells expressing GFP-paxillin also exhibited a discontinuous peeling in which focal adhesions fractured above sites of strong attachment (when pressure peeled using a small-bore pipette). The peeling approaches described here lend insight into the contractile-adhesion balance and can be used to study the real-time dynamics of stressed adhesions through both physical detection and the use of GFP markers; the methods should prove useful in comparing normal versus dystrophic muscle cells.