Lüchtefeld, Ines and Pivkin, Igor V. and Gardini, Lucia and Zare-Eelanjegh, Elaheh and Gäbelein, Christoph and Ihle, Stephan J. and Reichmuth, Andreas M. and Capitanio, Marco and Martinac, Boris and Zambelli, Tomaso and Vassalli, Massimo (2024) Dissecting cell membrane tension dynamics and its effect on Piezo1-mediated cellular mechanosensitivity using force-controlled nanopipettes. Nature Methods, 21 (6). pp.1063-1073. ISSN 1548-7091
Full text not available from this repository.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.
| Item Type: | Article |
|---|---|
| Subjects: | R Medicine > R Medicine (General) |
| Depositing User: | Repository Administrator |
| Date Deposited: | 23 Dec 2024 04:20 |
| Last Modified: | 23 Dec 2024 04:20 |
| URI: | https://eprints.victorchang.edu.au/id/eprint/1563 |
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