%0 Journal Article
%T Microscale geometrical modulation of PIEZO1 mediated mechanosensing through cytoskeletal redistribution.
%A Wang HJ
%A Wang Y
%A Mirjavadi SS
%A Andersen T
%A Moldovan L
%A Vatankhah P
%A Russell B
%A Jin J
%A Zhou Z
%A Li Q
%A Cox CD
%A Su QP
%A Ju LA
%J Nat Commun
%V 15
%N 1
%D 2024 Jun 29
%M 38951553
%F 17.694
%R 10.1038/s41467-024-49833-6
%X 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.