Stimuli-responsive microstructures are critical to create adaptable systems in soft robotics and biosciences. For such applications, the materials must be compatible with aqueous environments and enable the manufacturing of three-dimensional structures. Poly(N-isopropylacrylamide) (pNIPAM) is a well-established polymer, exhibiting a substantial response to changes in temperature close to its lower critical solution temperature. To create complex actuation patterns, materials that react differently with respect to a stimulus are required. Here, we introduce functional three-dimensional hetero-microstructures based on pNIPAM. By variation of the local exposure dose in three-dimensional laser lithography, we demonstrate that the material parameters can be altered on demand in a single resist formulation. We explore this concept for sophisticated three-dimensional architectures with large-amplitude and complex responses. The experimental results are consistent with numerical calculations, able to predict the actuation response. Furthermore, a spatially controlled response is achieved by inducing a local temperature increase by two-photon absorption of focused light.

Myotube formation by fusion of myoblasts and subsequent elongation of the syncytia is essential for skeletal muscle formation. However, molecules that regulate myotube formation remain elusive. Here we identify PIEZO1, a mechanosensitive Ca²⁺ channel, as a key regulator of myotube formation. During myotube formation, phosphatidylserine, a phospholipid that resides in the inner leaflet of the plasma membrane, is transiently exposed to cell surface and promotes myoblast fusion. We show that cell surface phosphatidylserine inhibits PIEZO1 and that the inward translocation of phosphatidylserine, which is driven by the phospholipid flippase complex of ATP11A and CDC50A, is required for PIEZO1 activation. PIEZO1-mediated Ca²⁺ influx promotes RhoA/ROCK-mediated actomyosin assemblies at the lateral cortex of myotubes, thus preventing uncontrolled fusion of myotubes and leading to polarized elongation during myotube formation. These results suggest that cell surface flip-flop of phosphatidylserine acts as a molecular switch for PIEZO1 activation that governs proper morphogenesis during myotube formation.