Journal Article

Microscale geometrical modulation of PIEZO1 mediated mechanosensing through cytoskeletal redistribution

Li, Qing, Andersen, Tomas, Cox, Charles D., Zhou, Zijing, Vatankhah, Parham, Su, Qian Peter, Wang, Yao, Ju, Lining Arnold, Russell, Blake, Wang, Haoqing Jerry, Jin, Jasmine, Moldovan, Laura, Mirjavadi, Seyed Sajad

2024

Overview

The microgeometry of the cellular microenvironment profoundly impacts cellular behaviors, yet the link between it and the ubiquitously expressed mechanosensitive ion channel PIEZO1 remains unclear. Herein, we describe a fluorescent micropipette aspiration assay that allows for simultaneous visualization of intracellular calcium dynamics and cytoskeletal architecture in real-time, under varied micropipette geometries. By integrating elastic shell finite element analysis with fluorescent lifetime imaging microscopy and employing PIEZO1-specific transgenic red blood cells and HEK cell lines, we demonstrate a direct correlation between the microscale geometry of aspiration and PIEZO1-mediated calcium signaling. We reveal that increased micropipette tip angles and physical constrictions lead to a significant reorganization of F-actin, accumulation at the aspirated cell neck, and subsequently amplify the tension stress at the dome of the cell to induce more PIEZO1’s activity. Disruption of the F-actin network or inhibition of its mobility leads to a notable decline in PIEZO1 mediated calcium influx, underscoring its critical role in cellular mechanosensing amidst geometrical constraints. Cells sense mechanical force through PIEZO1 channels. Here, authors show that both microgeometry and aspiration force mediate the PIEZO1 activity, and also alter F-actin organization, further amplifying PIEZO1 activity along membrane.

• Publisher: Nature Publishing Group UK,Nature Publishing Group,Nature Portfolio
• Subject: 13/106 / 14/19 / 14/34 / 14/63 / 631/57/2270/1140 / 631/80/128 / 631/80/79/2066 / 639/166/985 / 64/86 / 9/74 / 96/95 / Actin / Actins - metabolism / Amplification / Animals / Biomechanics / Biomedical engineering / Blood / Calcium / Calcium (blood) / Calcium (intracellular) / Calcium - metabolism / Calcium channels / Calcium imaging / Calcium influx / Calcium Signaling - physiology / Calcium signalling / Cell lines / Cytoskeleton / Cytoskeleton - metabolism / Elastic shells / Engineering / Erythrocytes / Finite Element Analysis / Finite element method / Fluorescence / HEK293 Cells / Humanities and Social Sciences / Humans / Intracellular signalling / Ion channels / Ion Channels - metabolism / Mechanotransduction, Cellular / Microenvironments / Microscopy, Fluorescence - methods / multidisciplinary / Real time / Science / Science (multidisciplinary)