Adherent cell remodeling on micropatterns is modulated by Piezo1 channels

Mohammad Reza Bahrani Fard,Frederick Sachs,Susan Z Hua,Katie Munechika,Deekshitha Jetta

doi:10.1038/s41598-021-84427-y

Mohammad Reza Bahrani Fard, Frederick Sachs + Show 3 more

Open Access

https://doi.org/10.1038/s41598-021-84427-y

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Abstract

Adherent cells utilize local environmental cues to make decisions on their growth and movement. We have previously shown that HEK293 cells grown on the fibronectin stripe patterns were elongated. Here we show that Piezo1 function is involved in cell spreading. Piezo1 expressing HEK cells plated on fibronectin stripes elongated, while a knockout of Piezo1 eliminated elongation. Inhibiting Piezo1 conductance using GsMTx4 or Gd3+ blocked cell spreading, but the cells grew thin tail-like extensions along the patterns. Images of GFP-tagged Piezo1 showed plaques of Piezo1 moving to the extrusion edges, co-localized with focal adhesions. Surprisingly, in non-spreading cells Piezo1 was located primarily on the nuclear envelope. Inhibiting the Rho-ROCK pathway also reversibly inhibited cell extension indicating that myosin contractility is involved. The growth of thin extrusion tails did not occur in Piezo1 knockout cells suggesting that Piezo1 may have functions besides acting as a cation channel.

Highlights

Adherent cells utilize local environmental cues to make decisions on their growth and movement
We show that Piezo[1] channels are essential for cell spreading on micro-patterns and knockout of Piezo[1] eliminates cell expansion
Expressing HEK cell line, HEK-hP1, we show that Piezo[1] proteins co-locate with focal adhesion complexes at the tips of spreading cells and this process involves myosin-II contractility via a Rho-ROCK pathway

Summary

Introduction

Adherent cells utilize local environmental cues to make decisions on their growth and movement. The growth of thin extrusion tails did not occur in Piezo[1] knockout cells suggesting that Piezo[1] may have functions besides acting as a cation channel. Cells detect and respond to mechanical cues with changes in shape, morphology, proliferation, and motility. This process is commonly thought to be mediated via adhesion proteins such as integrins at the adhesion complex[1,2,3]. Piezo[1] utilizes local environmental mechanical cues to regulate stem cell d ifferentiation[14]. Inhibiting Piezo[1] conductance with drugs leaves the cells with extended thin tail-like features, suggesting the Piezo[1] may have additional roles other than mediating Ca2+ signaling. Using a cloned Piezo[1]

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