Probing Single-Cell Mechanical Allostasis Using Ultrasound Tweezers
Probing Single-Cell Mechanical Allostasis Using Ultrasound Tweezers
复制标题
使用超声镊子探测单细胞机械动态平衡
DOI：
10.1007/s12195-019-00578-z
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发表时间：
2019
影响因子：
2.8
通讯作者：
Chen, Weiqiang
中科院分区：
文献类型：
--
作者：
Qian, Weiyi;Chen, Weiqiang
细胞对外界应激的反应是通过改变细胞的形态、代谢活动和功能，以机械地适应动态的局部环境。为了探索细胞变稳态中的机械转导，我们应用了一种集成的微机械系统，该系统结合了基于“超声镊子”的机械应力源和基于Förster共振能量转移（FRET）的分子力生物传感器，称为“辅肌动蛋白-sstFRET”，以监测原位单细胞对真实的瞬时刺激的变稳态。10-s瞬态超声脉冲，以声学激发与细胞膜结合的脂质包封的微泡，并通过RGD-整联蛋白连接向细胞施加皮科至纳牛顿范围的力。肌动蛋白sstFRET分子传感器，它从事肌动蛋白应力纤维在活细胞中，是用来映射实时肌动球蛋白力随时间的动态。然后，通过分析Ca 2+内流、肌动球蛋白细胞骨架（CSK）活性和GTdR RhoA信号传导的动态来检查机械敏感性行为，以定义单细胞机械别稳态。单个血管平滑肌细胞观察到在30分钟内以双相机械非稳态方式重塑自身，导致它们调整其收缩性和肌动球蛋白活性。强调CSK平衡在细胞机械别稳态中的重要作用的细胞机制包括Ca 2+内流、肌动球蛋白CSK和收缩的重塑以及GT3 RhoA信号传导。机械allostasis被观察到损害VSMCs与II型糖尿病（T2 DM），这可能会加强allostatic适应不良。ConclusionsBy整合工具，同时允许本地化的机械扰动和地图肌动球蛋白力，我们揭示了不同的细胞机械allostasis配置文件在我们的微机械系统。我们对细胞机械变稳态和适应不良的研究结果为机械表型细胞提供了潜力，以揭示其致病背景及其生物物理介质，这些介质是糖尿病，高血压或衰老等多病因疾病的基础。
IntroductionIn response to external stress, cells alter their morphology, metabolic activity, and functions to mechanically adapt to the dynamic, local environment through cell allostasis. To explore mechanotransduction in cellular allostasis, we applied an integrated micromechanical system that combines an ‘ultrasound tweezers’-based mechanical stressor and a Förster resonance energy transfer (FRET)-based molecular force biosensor, termed “actinin-sstFRET,” to monitorin situsingle-cell allostasis in response to transient stimulation in real time.MethodsThe ultrasound tweezers utilize 1 Hz, 10-s transient ultrasound pulses to acoustically excite a lipid-encapsulated microbubble, which is bound to the cell membrane, and apply a pico- to nano-Newton range of forces to cells through an RGD-integrin linkage. The actinin-sstFRET molecular sensor, which engages the actin stress fibers in live cells, is used to map real-time actomyosin force dynamics over time. Then, the mechanosensitive behaviors were examined by profiling the dynamics in Ca2+influx, actomyosin cytoskeleton (CSK) activity, and GTPase RhoA signaling to define a single-cell mechanical allostasis.ResultsBy subjecting a 1 Hz, 10-s physical stress, single vascular smooth muscle cells (VSMCs) were observed to remodeled themselves in a biphasic mechanical allostatic manner within 30 min that caused them to adjust their contractility and actomyosin activities. The cellular machinery that underscores the vital role of CSK equilibrium in cellular mechanical allostasis, includes Ca2+influx, remodeling of actomyosin CSK and contraction, and GTPase RhoA signaling. Mechanical allostasis was observed to be compromised in VSMCs from patients with type II diabetes mellitus (T2DM), which could potentiate an allostatic maladaptation.ConclusionsBy integrating tools that simultaneously permit localized mechanical perturbation and map actomyosin forces, we revealed distinct cellular mechanical allostasis profiles in our micromechanical system. Our findings of cell mechanical allostasis and maladaptation provide the potential for mechanophenotyping cells to reveal their pathogenic contexts and their biophysical mediators that underlie multi-etiological diseases such as diabetes, hypertension, or aging.
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影响因子：
3.4
作者：
Chen, Di;Sun, Yubing;Fu, Jianping
通讯作者：
Fu, Jianping
溶血磷脂酸在破坏内皮细胞-细胞粘附中的新作用
DOI：
--
发表时间：
2007
期刊：
影响因子：
--
作者：
Tsuda S.;et al.;田中 将之
通讯作者：
田中 将之
影响因子：
6.1
作者：
Mann JM;Lam RH;Weng S;Sun Y;Fu J
通讯作者：
Fu J
DOI：
--
发表时间：
--
期刊：
--
影响因子：
--
作者：
F. Chowdhury;S. Na;Dong Li;Y. Poh;Tetsuya S. Tanaka;Fei Wang;Ningwang
通讯作者：
F. Chowdhury;S. Na;Dong Li;Y. Poh;Tetsuya S. Tanaka;Fei Wang;Ningwang
影响因子：
13.3
作者：
Chen, Weiqiang;Allen, Steven G.;Fu, Jianping
通讯作者：
Fu, Jianping