Real-time monitoring of cell protrusion dynamics by impedance responses.
Real-time monitoring of cell protrusion dynamics by impedance responses.
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DOI：
10.1038/srep10206
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发表时间：
2015-05-15
影响因子：
4.6
通讯作者：
Primo L
中科院分区：
文献类型：
--
作者：
Gagliardi PA;Puliafito A;di Blasio L;Chianale F;Somale D;Seano G;Bussolino F;Primo L
细胞突起是参与基本过程的高度动态结构，包括细胞迁移和侵袭。对于迁移的细胞，其前缘必须形成突起，然后粘附或缩回。突出和缩回的空间和时间协调尚未完全理解。突起动力学的研究主要依赖于经常与荧光标记相结合的活体显微镜。在这里，我们报告了使用替代的，无标记的，定量和快速的测定，以分析突起动态的细胞群体的基础上，通过电子传感器的实时记录的细胞活性。将细胞接种在覆盖有电极的平板上，它们的形状变化映射到测量的阻抗变化。在生长因子刺激后，阻抗由于扩张活动而增加，并且在收缩后降低。与基于显微镜的方法相比，阻抗测量适用于不同细胞系、生长因子和化合物的高通量研究。我们提供的数据表明，该测定有助于剖析控制粘着突起的生化信号传导途径。事实上，我们表明，突起阶段是持续的肌动蛋白聚合，直接驱动的生长因子刺激。相反，收缩主要依赖于肌球蛋白的作用，指出肌球蛋白在板状伪足收缩中的关键作用。
Cellular protrusions are highly dynamic structures involved in fundamental processes, including cell migration and invasion. For a cell to migrate, its leading edge must form protrusions, and then adhere or retract. The spatial and temporal coordination of protrusions and retraction is yet to be fully understood. The study of protrusion dynamics mainly relies on live-microscopy often coupled to fluorescent labeling. Here we report the use of an alternative, label-free, quantitative and rapid assay to analyze protrusion dynamics in a cell population based on the real-time recording of cell activity by means of electronic sensors. Cells are seeded on a plate covered with electrodes and their shape changes map into measured impedance variations. Upon growth factor stimulation the impedance increases due to protrusive activity and decreases following retraction. Compared to microscopy-based methods, impedance measurements are suitable to high-throughput studies on different cell lines, growth factors and chemical compounds. We present data indicating that this assay lends itself to dissect the biochemical signaling pathways controlling adhesive protrusions. Indeed, we show that the protrusion phase is sustained by actin polymerization, directly driven by growth factor stimulation. Contraction instead mainly relies on myosin action, pointing at a pivotal role of myosin in lamellipodia retraction.
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影响因子：
4.8
作者：
Debnath, J;Muthuswamy, SK;Brugge, JS
通讯作者：
Brugge, JS
影响因子：
3.2
作者：
Heinrich, Doris;Youssef, Simon;Gerisch, Guenther
通讯作者：
Gerisch, Guenther
影响因子：
64.8
作者：
Keren, Kinneret;Pincus, Zachary;Theriot, Julie A.
通讯作者：
Theriot, Julie A.
DOI：
10.1083/jcb.119.5.1219
发表时间：
1992-12
期刊：
The Journal of cell biology
影响因子：
--
作者：
Lewis AK;Bridgman PC
通讯作者：
Bridgman PC
DOI：
10.1007/978-1-61779-108-6_6
发表时间：
2011-01-01
期刊：
MAMMALIAN CELL VIABILITY: METHODS AND PROTOCOLS
影响因子：
--
作者：
Ke, Ning;Wang, Xiaobo;Abassi, Yama A.
通讯作者：
Abassi, Yama A.