Abstract
Piezo1 is the stretch activated Ca2+ channel in red blood cells that mediates homeostatic volume control. Here, we study the organization of Piezo1 in red blood cells using a combination of super-resolution microscopy techniques and electron microscopy. Piezo1 adopts a non-uniform distribution on the red blood cell surface, with a bias toward the biconcave ‘dimple’. Trajectories of diffusing Piezo1 molecules, which exhibit confined Brownian diffusion on short timescales and hopping on long timescales, also reflect a bias toward the dimple. This bias can be explained by ‘curvature coupling’ between the intrinsic curvature of the Piezo dome and the curvature of the red blood cell membrane. Piezo1 does not form clusters with itself, nor does it colocalize with F-actin, Spectrin, or the Gardos channel. Thus, Piezo1 exhibits the properties of a force-through-membrane sensor of curvature and lateral tension in the red blood cell.
Statistical Analyses
For all comparative analyses n > 10 and the mean and standard deviation is given.
The one-way analysis of variance (ANOVA) with Tukeys post hoc analysis is used to test statistical significance.
Ethics
All animal procedures were reviewed and approved by the Institutional Animal Care and Use Committee at The Rockefeller University, protocol # 22056.
