Zyxin is involved in regulation of mechanotransduction in arteriole smooth muscle cells.

Zyxin is involved in regulation of mechanotransduction in arteriole smooth muscle cells.
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DOI:
10.3389/fphys.2012.00472
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发表时间:
2012
影响因子:
4
通讯作者:
Meininger GA
Meininger GA
中科院分区:
医学2区
文献类型:
--
作者:
Sun Z;Huang S;Li Z;Meininger GA

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Zyxin 是一种粘附斑蛋白,与细胞粘附和运动的调节有关,并被假设为整合素介导的机械力反应中的机械传感器。为了测试 zyxin 在微血管平滑肌细胞 (VSMC) 机械转导中的功能作用,我们利用原子力显微镜 (AFM) 通过细胞表面涂有 FN 的珠子诱导的纤连蛋白 (FN) 粘着斑向 VSMC 施加局部拉力。使用 AFM 施加力会导致 FN 珠粘着斑部位的 zyxin 积累增加,并伴随 VSMC 收缩反应。然而,通过使用 zyxin-shRNA 构建体减少 zyxin 表达消除了 VSMC 对 AFM 拉力的收缩反应,尽管 zyxin 沉默的 VSMC 在 AFM 粘附测定和细胞粘附测定中均表现出对 FN 的粘附增加。 zyxin 表达的减少显着损害了细胞扩散和肌动蛋白细胞骨架的重组,这可能表明机械力收缩反应丧失的可能潜在原因。与这些观察结果一致,在 zyxin 沉默的 VSMC 中,我们还观察到 Rac1 的表达减少,Rac1 在 VSMC 的肌动蛋白重组中发挥重要作用,但甲状腺受体相互作用蛋白 (TRIP6) 和 FAK 的表达增加,后者是促进细胞粘附的主要蛋白。总之,这些数据支持 zyxin 在 VSMC 对所施加的力进行机械响应的能力中发挥重要的促进作用。
Zyxin is a focal adhesion protein that has been implicated in the modulation of cell adhesion and motility, and is hypothesized to be a mechano-sensor in integrin-mediated responses to mechanical force. To test the functional role of zyxin in the mechanotransduction of microvascular smooth muscle cells (VSMC), we utilized atomic force microscopy (AFM) to apply localized pulling forces to VSMC through a fibronectin (FN) focal adhesion induced by a FN-coated bead on cell surface. Application of force with the AFM induced an increase of zyxin accumulation at the site of the FN-bead focal adhesion that accompanied the VSMC contractile response. Whereas, reduction of zyxin expression by using a zyxin-shRNA construct abolished the VSMC contractile response to AFM pulling forces, even though the zyxin-silenced VSMCs displayed increased adhesion to FN in both AFM adhesion assays and cell adhesion assays. The reduced zyxin expression significantly impaired cell spreading and reorganization of the actin cytoskeleton that could indicate a possible underlying reason for the loss of a contractile response to mechanical force. Consistent with these observations, in zyxin-silenced VSMC, we also observed a reduced expression of Rac1, which plays an important role in the actin reorganization in VSMC, but increased thyroid receptor-interacting proteins (TRIP6) and FAK expression, the latter being a major protein that promote cell adhesion. In conclusion, these data support an important enabling role for zyxin in VSMCs ability to mechanically respond to applied force.
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