Micropatterning as a tool to identify regulatory triggers and kinetics of actin-mediated endothelial mechanosensing

Abstract

Developmental processes such as angiogenesis are associated to a constant remodeling of the actin cytoskeleton in response to different mechanical stimuli. The mechanosensitive transcription factors MRTF and YAP are important mediators of this challenging adaptation process. However, it is yet unknown whether both pathways respond identical or divergent to a given microenvironmental guidance cue. Here, we use a micropatterning approach to dissect single aspects of cellular behavior in a spatiotemporally controllable setting. Using the exemplary process of angiogenesis, we show that cell-cell contacts and adhesive surface area are shared regulatory parameters of MRTF and YAP on rigid 2D surfaces. By analyzing MRTF and YAP under laminar flow conditions and during cell migration on dumbbell-shaped microstructures, we demonstrate that they exhibit different translocation kinetics. In conclusion, our work promotes the application of micropatterning techniques as a cell-biological tool to study mechanosensitive signaling in the context of angiogenesis.