Single-particle tracking reveals heterogeneous PIEZO1 diffusion

Area of Science:

• Cell Biology
• Biophysics
• Membrane Protein Dynamics

Background:

• The mechanically activated ion channel PIEZO1 plays a crucial role in various physiological processes.
• PIEZO1 is activated by mechanical cues and its mobility in the plasma membrane is essential for its function.

Purpose of the Study:

• To investigate the diffusion dynamics of endogenous PIEZO1 in live cells.
• To determine how plasma membrane composition and channel activation affect PIEZO1 mobility.

Main Methods:

• Single-particle tracking (SPT) of tdTomato-tagged PIEZO1 using total internal reflection fluorescence microscopy.
• Manipulation of membrane cholesterol levels and addition of margaric acid.
• Treatment with PIEZO1 agonist Yoda1 and inhibitor GsMTx-4.

Main Results:

• PIEZO1 displayed heterogeneous diffusion, categorized as mobile and immobile subpopulations.
• Cholesterol depletion and Yoda1 treatment increased PIEZO1 mobility.
• GsMTx-4 treatment and cholesterol supplementation decreased PIEZO1 mobility.
• Mobile PIEZO1 puncta exhibited anomalous subdiffusion.

Conclusions:

• Plasma membrane composition and PIEZO1 activation state critically regulate its diffusion.
• Understanding PIEZO1 mobility is key to elucidating its role in cellular mechanotransduction.
• These findings provide a foundation for studying how cellular processes influence PIEZO1 activity.