Abstract
Epithelial cells from diverse systems assemble apical microvilli to mediate biochemical and physical interactions with the external environment. Despite the central importance of these actin-based protrusions in epithelial physiology, how microvilli grow during differentiation remains poorly understood. Here, we report that an epithelial cell's potential for growing microvilli of normal size is limited by an adjacent actin-dependent process: clathrin-mediated endocytosis. Time-lapse imaging of individual microvillar growth events revealed tight spatial and temporal coupling to sites of apical clathrin-mediated endocytosis. Moreover, ultrastructural characterization of differentiating epithelia showed that most nascent microvilli are in contact with an endocytic pit. Finally, inhibition of the Arp2/3 complex, which drives actin polymerization on coated pits, significantly reduced the accumulation of new microvilli on the cell surface. We conclude that clathrin-mediated endocytosis and its associated Arp2/3-based actin nucleation activity control the timing and location of microvillar growth as well as the dimensions of the resulting protrusions.