Förster Resonance Energy Transfer-Based Single-Cell Imaging Reveals Piezo1-Induced Ca2+ Flux Mediates Membrane Ruffling and Cell Survival

Heon-Su Kim; Jung-Soo Suh; Yoon-Kwan Jang; Sang-Hyun Ahn; Gyu-Ho Choi; Jin-Young Yang; Gah-Hyun Lim; Youngmi Jung; Jie Jiang; Jie Sun; Myungeun Suk; Yingxiao Wang; Tae-Jin Kim

doi:10.3389/fcell.2022.865056

Förster Resonance Energy Transfer-Based Single-Cell Imaging Reveals Piezo1-Induced Ca²⁺ Flux Mediates Membrane Ruffling and Cell Survival

Front Cell Dev Biol. 2022 May 13:10:865056. doi: 10.3389/fcell.2022.865056. eCollection 2022.

Authors

Heon-Su Kim^{1

2}, Jung-Soo Suh¹, Yoon-Kwan Jang¹, Sang-Hyun Ahn¹, Gyu-Ho Choi¹, Jin-Young Yang¹, Gah-Hyun Lim¹, Youngmi Jung¹, Jie Jiang³, Jie Sun³, Myungeun Suk⁴, Yingxiao Wang⁵, Tae-Jin Kim^{1

2

6}

Affiliations

¹ Department of Integrated Biological Science, Pusan National University, Pusan, South Korea.
² Institute of Systems Biology, Pusan National University, Pusan, South Korea.
³ Department of Cell Biology, School of Medicine, Zhejiang University, Hangzhou, China.
⁴ Department of Mechanical Engineering, Dong-Eui University, Pusan, South Korea.
⁵ Department of Bioengineering, Institute of Engineering in Medicine, University of California, San Diego, La Jolla, CA, United States.
⁶ Department of Biological Sciences, Pusan National University, Pusan, South Korea.

Abstract

A mechanosensitive ion channel, Piezo1 induces non-selective cation flux in response to various mechanical stresses. However, the biological interpretation and underlying mechanisms of cells resulting from Piezo1 activation remain elusive. This study elucidates Piezo1-mediated Ca²⁺ influx driven by channel activation and cellular behavior using novel Förster Resonance Energy Transfer (FRET)-based biosensors and single-cell imaging analysis. Results reveal that extracellular Ca²⁺ influx via Piezo1 requires intact caveolin, cholesterol, and cytoskeletal support. Increased cytoplasmic Ca²⁺ levels enhance PKA, ERK, Rac1, and ROCK activity, which have the potential to promote cancer cell survival and migration. Furthermore, we demonstrate that Piezo1-mediated Ca²⁺ influx upregulates membrane ruffling, a characteristic feature of cancer cell metastasis, using spatiotemporal image correlation spectroscopy. Thus, our findings provide new insights into the function of Piezo1, suggesting that Piezo1 plays a significant role in the behavior of cancer cells.

Keywords: FRET; Piezo1; biosensors; calcium; membrane ruffling.

Abstract

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