英国格拉斯哥大学Massimo Vassalli等研究人员合作利用力控纳米吸管剖析细胞膜张力动态及其对Piezo1介导的细胞机械敏感性的影响。相关论文于2024年5月27日在线发表在《自然—方法学》杂志上。

研究人员报告了一种基于力控纳米吸管的方法，该方法结合了流体力显微镜和荧光成像技术，可精确操纵细胞膜张力，同时监测其对单细胞机械敏感性的影响。力控纳米吸管可以控制施加在细胞皮层上的压痕力以及施加在质膜上的吸力。研究表明，这种装置可用于通过钙成像同时监测Piezo1机械敏感离子通道的激活。

此外，研究人员还利用荧光膜张力探针Flipper-TR评估了张力传播的时空行为，并利用分子动力学建模对其进行了进一步剖析。最后，研究人员证明了吸气和压痕对细胞机械生物学机制的独立作用，压痕在膜中诱导局部预张力，而膜张力通过与细胞骨架的连接保持局限。

据了解，细胞膜张力是细胞对物理刺激做出反应的能力，而细胞膜张力的动态及其在机械传感中的作用仍不完全清楚，这主要是由于缺乏适当的工具。

附：英文原文

Title: Dissecting cell membrane tension dynamics and its effect on Piezo1-mediated cellular mechanosensitivity using force-controlled nanopipettes

Author: Lchtefeld, Ines, Pivkin, Igor V., Gardini, Lucia, Zare-Eelanjegh, Elaheh, Gbelein, Christoph, Ihle, Stephan J., Reichmuth, Andreas M., Capitanio, Marco, Martinac, Boris, Zambelli, Tomaso, Vassalli, Massimo

Issue&Volume: 2024-05-27

Abstract: The dynamics of cellular membrane tension and its role in mechanosensing, which is the ability of cells to respond to physical stimuli, remain incompletely understood, mainly due to the lack of appropriate tools. Here, we report a force-controlled nanopipette-based method that combines fluidic force microscopy with fluorescence imaging for precise manipulation of the cellular membrane tension while monitoring the impact on single-cell mechanosensitivity. The force-controlled nanopipette enables control of the indentation force imposed on the cell cortex as well as of the aspiration pressure applied to the plasma membrane. We show that this setup can be used to concurrently monitor the activation of Piezo1 mechanosensitive ion channels via calcium imaging. Moreover, the spatiotemporal behavior of the tension propagation is assessed with the fluorescent membrane tension probe Flipper-TR, and further dissected using molecular dynamics modeling. Finally, we demonstrate that aspiration and indentation act independently on the cellular mechanobiological machinery, that indentation induces a local pre-tension in the membrane, and that membrane tension stays confined by links to the cytoskeleton.

DOI: 10.1038/s41592-024-02277-8

Source: https://www.nature.com/articles/s41592-024-02277-8