Microscale geometrical modulation of PIEZO1 mediated mechanosensing through cytoskeletal redistribution

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

doi:10.1038/s41467-024-49833-6

Microscale geometrical modulation of PIEZO1 mediated mechanosensing through cytoskeletal redistribution

Nat Commun. 2024 Jun 29;15(1):5521. doi: 10.1038/s41467-024-49833-6.

Authors

Haoqing Jerry Wang^{1

2

3}, Yao Wang¹, Seyed Sajad Mirjavadi⁴, Tomas Andersen¹, Laura Moldovan^{1

2

3}, Parham Vatankhah¹, Blake Russell¹, Jasmine Jin¹, Zijing Zhou⁵, Qing Li⁴, Charles D Cox^{5

6}, Qian Peter Su^{7

8}, Lining Arnold Ju^{9

10

11

12}

Affiliations

¹ School of Biomedical Engineering, The University of Sydney, Darlington, NSW, 2008, Australia.
² Charles Perkins Centre, The University of Sydney, Camperdown, NSW, 2006, Australia.
³ Heart Research Institute, Camperdown, Newtown, NSW, 2042, Australia.
⁴ School of Aerospace, Mechanical and Mechatronic Engineering, The University of Sydney, Darlington, NSW, 2008, Australia.
⁵ Molecular Cardiology and Biophysics Division, Victor Chang Cardiac Research Institute, Sydney, NSW, 2010, Australia.
⁶ Faculty of Medicine, St. Vincent's Clinical School, University of New South Wale, Sydney, NSW, 2010, Australia.
⁷ Heart Research Institute, Camperdown, Newtown, NSW, 2042, Australia. [email protected].
⁸ School of Biomedical Engineering, University of Technology Sydney, Sydney, NSW, 2007, Australia. [email protected].
⁹ School of Biomedical Engineering, The University of Sydney, Darlington, NSW, 2008, Australia. [email protected].
¹⁰ Charles Perkins Centre, The University of Sydney, Camperdown, NSW, 2006, Australia. [email protected].
¹¹ Heart Research Institute, Camperdown, Newtown, NSW, 2042, Australia. [email protected].
¹² The University of Sydney Nano Institute (Sydney Nano), The University of Sydney, Camperdown, NSW, 2006, Australia. [email protected].

Abstract

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.

MeSH terms

Actins* / metabolism
Animals
Calcium Signaling / physiology
Calcium* / metabolism
Cytoskeleton* / metabolism
Finite Element Analysis
HEK293 Cells
Humans
Ion Channels* / metabolism
Mechanotransduction, Cellular*
Microscopy, Fluorescence / methods

Substances

Ion Channels
PIEZO1 protein, human
Actins
Calcium
Piezo1 protein, mouse

Abstract

MeSH terms

Substances

Grants and funding