BPS2026 – Engineering compliant mechanical environments to probe crosslinker behavior in cytoskeletal ensembles

The cytoskeleton is a dynamic, mechanoresponsive network composed of actin filaments and microtubules and is vital for structural support, cell motility, force generation, and adhesion. Anillin is a diffusive crosslinker that connects actin filaments directly to microtubules, generates force through entropic expansion and adapts to different mechanical environments. Anillin’s behavior and ability to crosslink actin filaments and microtubules is influenced by the mechanical properties of the surrounding cytoskeleton environment, particularly surface compliance. However, the extent to which extracellular and intracellular mechanical cues influence anillin’s behavior at the molecular level remains poorly defined. Our study aims to develop a controlled mechanical environment through layer-by-layer (LbL) thin film assembly to construct a tunable mechanical platform composed of collagen/heparin bilayers, supporting a more compliant microenvironment. We characterize the properties of the films using a quartz crystal microbalance with dissipation monitoring (QCM-D) to assess nanoscale changes in mass and viscoelasticity and use optical tweezers to probe the force-generating behavior of anillin crosslinkers in actin-microtubule networks working on films. Results suggest that a compliant environment will impact anillin’s ability to generate force through entropic expansion. Systematic investigation of engineered mechanical environments and their effects on crosslinker behavior could yield new insight into force feedback loops that regulate cytoskeletal dynamics.