title: Synaptophysin 1 Clears Synaptobrevin 2 from the Presynaptic Active Zone to Prevent Short-Term Depression.
abstract: Release site clearance is an important process during synaptic vesicle (SV) recycling. However, little is known about its molecular mechanism. Here we identify self-assembly of exocytosed Synaptobrevin 2 (Syb2) and Synaptophysin 1 (Syp1) by homo- and hetero-oligomerization into clusters as key mechanisms mediating release site clearance for preventing cis-SNARE complex formation at the active zone (AZ). In hippocampal neurons from Syp1 knockout mice, neurons expressing a monomeric Syb2 mutant, or after acute block of the ATPase N-ethylmaleimide-sensitive factor (NSF), responsible for cis-SNARE complex disassembly, we found strong frequency-dependent short-term depression (STD), whereas retrieval of Syb2 by compensatory endocytosis was only affected weakly. Defects in Syb2 endocytosis were stimulus- and frequency-dependent, indicating that Syp1 is not essential for Syb2 retrieval, but for its efficient clearance upstream of endocytosis. Our findings identify an SV protein as a release site clearance factor.
authors: Department of Cellular Biophysics, Institute of Medical Physics and Biophysics, University of Münster, 48149 Münster, Germany; Cells in Motion, International Max Planck Research School Graduate Program, Schlossplatz 5, 48149 Münster, Germany.
authors: Rajit
authors: R
authors: Rajappa
authors: Department of Cellular Biophysics, Institute of Medical Physics and Biophysics, University of Münster, 48149 Münster, Germany.
authors: Anne
authors: A
authors: Gauthier-Kemper
authors: Department of Cellular Biophysics, Institute of Medical Physics and Biophysics, University of Münster, 48149 Münster, Germany.
authors: Daniel
authors: D
authors: Böning
authors: Department of Cellular Biophysics, Institute of Medical Physics and Biophysics, University of Münster, 48149 Münster, Germany; Fluorescence Microscopy Facility Münster, Center for Nanotechnology, 48149 Münster, Germany.
authors: Jana
authors: J
authors: Hüve
authors: Department of Cellular Biophysics, Institute of Medical Physics and Biophysics, University of Münster, 48149 Münster, Germany; Fluorescence Microscopy Facility Münster, Center for Nanotechnology, 48149 Münster, Germany; Cluster of Excellence EXC 1003, Cells in Motion, 48149 Münster, Germany. Electronic address: klingauf@uni-muenster.de.
authors: Jürgen
authors: J
authors: Klingauf
mesh_terms: D000818:Animals; D000831:Animals, Newborn; D004705:Endocytosis / Q000502:physiology; D005089:Exocytosis / Q000502:physiology; D005786:Gene Expression Regulation; D057809:HEK293 Cells; D006624:Hippocampus / Q000166:cytology / Q000378:metabolism; D006801:Humans; D051379:Mice; D018345:Mice, Knockout; D052066:N-Ethylmaleimide-Sensitive Proteins / Q000235:genetics / Q000378:metabolism; D009433:Neural Inhibition / Q000502:physiology*; D009474:Neurons / Q000166:cytology / Q000378:metabolism*; D016716:PC12 Cells; D017729:Presynaptic Terminals / Q000378:metabolism* / Q000648:ultrastructure; D061251:Primary Cell Culture; D055503:Protein Multimerization; D051381:Rats; D050600:SNARE Proteins / Q000235:genetics / Q000378:metabolism; D013572:Synaptic Vesicles / Q000378:metabolism* / Q000648:ultrastructure; D016708:Synaptophysin / Q000235:genetics* / Q000378:metabolism; D050988:Vesicle-Associated Membrane Protein 2 / Q000235:genetics* / Q000378:metabolism
chemical_list: D050600:SNARE Proteins; D016708:Synaptophysin; C569016:Syp protein, rat; C494706:Vamp2 protein, rat; D050988:Vesicle-Associated Membrane Protein 2; D052066:N-Ethylmaleimide-Sensitive Proteins