Stochastic contraction of myosin minifilaments drives evolution of microridge protrusion patterns in epithelial cells

Actin-based protrusions vary in morphology, stability, and arrangement on cell surfaces. Microridges are laterally-elongated protrusions on mucosal epithelial cells, where they are arranged in evenly spaced, maze-like patterns that dynamically remodel by fission and fusion. To characterize how microridges form their highly ordered patterns, and investigate the mechanisms driving fission and fusion, we imaged microridges in the maturing skin of zebrafish larvae. After their initial development, microridge spacing and alignment became increasingly well ordered. Imaging F-actin and Non-Muscle Myosin II (NMII) revealed that microridge fission and fusion were associated with local NMII activity in the apical cortex. Inhibiting NMII blocked fission and fusion rearrangements, reduced microridge density, and altered microridge spacing. High-resolution imaging allowed us to image individual NMII minifilaments in the apical cortex of cells in live animals, revealing that minifilaments are tethered to protrusions and often connected adjacent microridges. NMII minifilaments connecting the ends of microridges fused them together, whereas minifilaments oriented perpendicular to microridges severed them or pulled them closer together. These findings demonstrate that as cells mature, cortical NMII activity orchestrates a microridge remodeling process that creates an increasingly orderly microridge arrangement.