Impairing actin filament or syndapin functions promotes accumulation of clathrin-coated vesicles at the apical plasma membrane of acinar epithelial cells |
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| Authors: | Da Costa Silvia R Sou Eunbyul Xie Jiansong Yarber Francie A Okamoto Curtis T Pidgeon Michael Kessels Michael M Mircheff Austin K Schechter Joel E Qualmann Britta Hamm-Alvarez Sarah F |
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| Affiliation: | Department of Pharmaceutical Sciences, University of Southern California, Los Angeles, California 90033, USA. |
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| Abstract: | In this article, we investigate the contributions of actin filaments and accessory proteins to apical clathrin-mediated endocytosis in primary rabbit lacrimal acini. Confocal fluorescence and electron microscopy revealed that cytochalasin D promoted apical accumulation of clathrin, alpha-adaptin, dynamin, and F-actin and increased the amounts of coated pits and vesicles at the apical plasma membrane. Sorbitol density gradient analysis of membrane compartments showed that cytochalasin D increased [14C]dextran association with apical membranes from stimulated acini, consistent with functional inhibition of apical endocytosis. Recombinant syndapin SH3 domains interacted with lacrimal acinar dynamin, neuronal Wiskott-Aldrich Syndrome protein (N-WASP), and synaptojanin; their introduction by electroporation elicited remarkable accumulation of clathrin, accessory proteins, and coated pits at the apical plasma membrane. These SH3 domains also significantly (p
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