title: Clathrin regenerates synaptic vesicles from endosomes.
abstract: Ultrafast endocytosis can retrieve a single, large endocytic vesicle as fast as 50-100 ms after synaptic vesicle fusion. However, the fate of the large endocytic vesicles is not known. Here we demonstrate that these vesicles transition to a synaptic endosome about one second after stimulation. The endosome is resolved into coated vesicles after 3 s, which in turn become small-diameter synaptic vesicles 5-6 s after stimulation. We disrupted clathrin function using RNA interference (RNAi) and found that clathrin is not required for ultrafast endocytosis but is required to generate synaptic vesicles from the endosome. Ultrafast endocytosis fails when actin polymerization is disrupted, or when neurons are stimulated at room temperature instead of physiological temperature. In the absence of ultrafast endocytosis, synaptic vesicles are retrieved directly from the plasma membrane by clathrin-mediated endocytosis. These results may explain discrepancies among published experiments concerning the role of clathrin in synaptic vesicle endocytosis.
authors.affiliation: Department of Biology and Howard Hughes Medical Institute, University of Utah, Salt Lake City, Utah 84112-0840, USA.
authors.forename: Shigeki
authors.initials: S
authors.lastname: Watanabe
authors.affiliation: Neuroscience Research Center Charité Universitätsmedizin Berlin, Berlin 10117, Germany.
authors.forename: Thorsten
authors.initials: T
authors.lastname: Trimbuch
authors.affiliation: Neuroscience Research Center Charité Universitätsmedizin Berlin, Berlin 10117, Germany.
authors.forename: Marcial
authors.initials: M
authors.lastname: Camacho-Pérez
authors.affiliation: 1] Neuroscience Research Center Charité Universitätsmedizin Berlin, Berlin 10117, Germany [2] German Center for Neurodegenerative Diseases (DZNE), 10117 Berlin, Germany.
authors.forename: Benjamin R
authors.initials: BR
authors.lastname: Rost
authors.affiliation: Neuroscience Research Center Charité Universitätsmedizin Berlin, Berlin 10117, Germany.
authors.forename: Bettina
authors.initials: B
authors.lastname: Brokowski
authors.affiliation: Neuroscience Research Center Charité Universitätsmedizin Berlin, Berlin 10117, Germany.
authors.forename: Berit
authors.initials: B
authors.lastname: Söhl-Kielczynski
authors.affiliation: Neuroscience Research Center Charité Universitätsmedizin Berlin, Berlin 10117, Germany.
authors.forename: Annegret
authors.initials: A
authors.lastname: Felies
authors.affiliation: Department of Biology and Howard Hughes Medical Institute, University of Utah, Salt Lake City, Utah 84112-0840, USA.
authors.forename: M Wayne
authors.initials: MW
authors.lastname: Davis
authors.affiliation: Neuroscience Research Center Charité Universitätsmedizin Berlin, Berlin 10117, Germany.
authors.forename: Christian
authors.initials: C
authors.lastname: Rosenmund
authors.affiliation: Department of Biology and Howard Hughes Medical Institute, University of Utah, Salt Lake City, Utah 84112-0840, USA.
authors.forename: Erik M
authors.initials: EM
authors.lastname: Jorgensen
mesh_terms: D000818:Animals; D002462:Cell Membrane / Q000378:metabolism; D002966:Clathrin / Q000378:metabolism*; D004705:Endocytosis; D011992:Endosomes / Q000378:metabolism*; D006801:Humans; D051379:Mice; D013572:Synaptic Vesicles / Q000378:metabolism*; D013696:Temperature
chemical_list: D002966:Clathrin