A Microfluidic Platform for Actin-Based Membrane Remodeling Reveals the Stabilizing Role of Branched Actin Networks on Lipid Microdomains

Abstract: Cell shape changes, essential for processes such as motility or division, are controlled by the actomyosin cortex that actively remodels biological membranes. Their mechanisms can be deciphered in_vitro using biomimetic reconstituted systems, such as giant unilamellar vesicles (GUVs) with controlled lipid composition coupled to reconstituted actin networks. These assays allow mimicking cell shape changes in controlled biochemical and biophysical environments. However, studying the dynamics of these shape changes on statistically significant populations of GUVs with the possibility to sequentially modify the protein composition of the assay is a major experimental challenge. To address these issues, a microfluidic approach is used to immobilize several dozens of isolated GUVs and monitor membrane and actin network evolution. The loading of the chamber with GUVs and actin is first characterized. Then, the actin-induced remodeling of populations of homogeneous and phase-separated GUVs is monitored and shows that actin networks prevent the coalescence of lipid microdomains and that, in return, the number of domains affects the actin network structure. This microfluidic-based experimental strategy, thus, allows for studying actin-induced membrane deformation in_vitro and can be adapted to other studies on membrane remodeling processes.