Piezo1 is a mechanically activated ion channel involved in sensing forces in various cell types and tissues. Cryo-electron microscopy has revealed that the Piezo1 structure is bowl-shaped and capable of inducing membrane curvature via its extended footprint, which indirectly suggests that Piezo1 ion channels may bias each other's spatial distribution and interact functionally. Here, we use cell-attached patch-clamp electrophysiology and pressure-clamp stimulation to functionally examine large numbers of membrane patches from cells expressing Piezo1 endogenously at low levels and cells overexpressing Piezo1 at high levels. Our data, together with stochastic simulations of Piezo1 spatial distributions, show that both at endogenous densities (1-2 channels/μm2), and at non-physiological densities (10-100 channels/μm2) predicted to cause substantial footprint overlap, Piezo1 density has no effect on its pressure sensitivity or open probability in the nominal absence of membrane tension. The results suggest that Piezo channels, at densities likely to be physiologically relevant, inherently behave as independent mechanotransducers. We propose that this property is essential for cells to transduce forces homogeneously across the entire cell membrane.
Keywords: Piezo1; clustering; cooperativity; force-gated ion channel; mechanotransduction; molecular biophysics; neuroscience; none; structural biology.
Cells can sense a range of mechanical forces both inside and outside the body, such as the stroke of a fingertip or the filling of a lung. Pores on the surface of the cell called Piezo channels open up in response to this pressure. This allows ions to flood in to the cell and trigger a series of biochemical reactions that alter the cell’s behavior. Piezo channels have a unique bowl-like structure that transforms the shape of the cell surface around them, potentially affecting how nearby proteins behave. Previous research had suggested that these channels might be unevenly distributed across the cell surface, and were predicted to modify each other’s behaviors when tightly packed together. This cooperative response would have a significant impact on how cells sense mechanical force. To investigate if this was the case, Lewis and Grandl studied a mouse cell called Neuro2A which naturally produces Piezo ion channels. In the experiment, pressure was applied to different parts of the cell and the electric current generated by ions moving across the surface was recorded: the higher the electrical activity, the more ion channels present. This showed that Piezo channels are randomly distributed across the cell surface and do not tend to cluster together. The same Neuro2A cells were then engineered to produce up to one hundred times more Piezo proteins. Despite the channels being more densely packed together, how they responded to mechanical force remained the same. These results suggest that Piezo channels act independently and are not influenced by close proximity to one another. Lewis and Grandl propose that this property may ensure that all parts of the cell surface react to mechanical force in the same way. Further work is needed to see if this finding applies to other cell types that produce Piezo proteins.
© 2021, Lewis and Grandl.