A new automated high-throughput approach to studying PIEZO channels
The discovery of PIEZO channels in 2010 marked a turning point in mechanobiology research, leading numerous teams to study the role of these channels in various physiological and pathological processes. Since then, numerous publications have confirmed the key role of PIEZO channels in mechanosensitivity across species. Dysfunctions in Piezo1 and Piezo2 have been found to be linked to various severe pathological conditions, from generalized lymphatic dysplasia and varicose vein disease to hereditary xerocytosis—a condition that affects red blood cells.
Studying PIEZO’s mechanical activation has traditionally been restricted to conventional manual patch clamp techniques combined with different mechanical stimulation methods, such as hitting cells with a stylus, applying fluid flow to cause shear stress, or applying pressure pulses to the patch pipette. Although these approaches have revealed much about the properties of PIEZO channels, they do not easily lend themselves to the rapid screening required for drug discovery. Thus, the lack of patch clamp systems capable of applying mechanical stimulation in an automated, high-throughput manner has been a bottleneck in drug development targeting PIEZO channels. Until now.
A recent study marks a breakthrough by presenting a scalable technique that allows for the investigation of PIEZO’s mechanical response using automated patch clamp (APC) technology. Researchers have optimized the SyncroPatch 384, a high-throughput APC system from Nanion, for high-speed solution application to cells, creating a robust method for studying PIEZO activity under mechanical stress. They showed that applying solutions on top of the cells at elevated pipetting flows (110 µl/s) effectively activates PIEZO1 channels on the SyncroPatch 384. The authors used this approach to explore differences in the responses of mouse and human PIEZO1 channels to mechanical and/or chemical (Yoda1) stimuli. Consistent with previous findings, hPIEZO1 exhibited faster activation and decay time courses of the currents compared to mPIEZO1, indicating that this new automated approach allows for the comparison and differentiation between two different PIEZO1 clones.
This new possibility of comparing and combining mechanical and chemical stimulation in a high-throughput automated patch clamp assay now greatly facilitates investigations into mechanically activated PIEZO channels and provides an important experimental tool both for fundamental studies and drug development.
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