Clathrin regenerates synaptic vesicles from endosomes

Shigeki Watanabe, Thorsten Trimbuch, Marcial Camacho-Pérez, Benjamin R. Rost, Bettina Brokowski, Berit Söhl-Kielczynski, Annegret Felies, M. Wayne Davis, Christian Rosenmund () and Erik M. Jorgensen ()
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Shigeki Watanabe: University of Utah
Thorsten Trimbuch: Neuroscience Research Center Charité Universitätsmedizin Berlin, Berlin 10117, Germany
Marcial Camacho-Pérez: Neuroscience Research Center Charité Universitätsmedizin Berlin, Berlin 10117, Germany
Benjamin R. Rost: Neuroscience Research Center Charité Universitätsmedizin Berlin, Berlin 10117, Germany
Bettina Brokowski: Neuroscience Research Center Charité Universitätsmedizin Berlin, Berlin 10117, Germany
Berit Söhl-Kielczynski: Neuroscience Research Center Charité Universitätsmedizin Berlin, Berlin 10117, Germany
Annegret Felies: Neuroscience Research Center Charité Universitätsmedizin Berlin, Berlin 10117, Germany
M. Wayne Davis: University of Utah
Christian Rosenmund: Neuroscience Research Center Charité Universitätsmedizin Berlin, Berlin 10117, Germany
Erik M. Jorgensen: University of Utah

Nature, 2014, vol. 515, issue 7526, 228-233

Abstract: 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.

Date: 2014
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DOI: 10.1038/nature13846

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