Abstract
Due to the long time required for de novo formation of deeply invaginated clathrin-coated pits that mostly appear at sites different from exocytosis before fission, clathrin-dependent vesicle recycling in secretory cells is usually regarded as a slow process. Here we show that regulated exocytosis in pancreatic beta cells is associated with glucose-dependent recruitment of clathrin and dynamin-1 puncta at the release sites shortly or long after fusion pore opens. The subsequent disassembly of these clathrin puncta mediates internalization of synaptotagmin VII clusters in the plasma membrane. The fast clathrin-dependent endocytosis contributes significantly to the total vesicle recycling process, and involves microtubule-dependent linear movement of preformed clathrin-coated pits to vesicle release sites prior to fission. These results have revealed an unexpected close spatio-temporal coupling of clathrin-dependent endocytosis to vesicle fusion in pancreatic beta cells, and highlighted a novel pathway to replenish the “readily retrieval pool” of clathrin that sustains fast clathrin-dependent endocytosis under intense stimulation.
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