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Updated: Apr 28, 2026

Measuring Synaptic Vesicle Endocytosis in Cultured Hippocampal Neurons
Published on: September 4, 2017
Clathrin/AP-2 mediate synaptic vesicle reformation from endosome-like vacuoles but are not essential for membrane
Natalia L Kononenko1, Dmytro Puchkov2, Gala A Classen1
1Leibniz Institut für Molekulare Pharmakologie (FMP) & Freie Universität Berlin, Robert-Roessle-Straße 10, 13125 Berlin, Germany; Charite Universitätsmedizin, NeuroCure Cluster of Excellence, Virchowweg 6, 10117 Berlin, Germany.
Abstract:
Neurotransmission depends on presynaptic membrane retrieval and local reformation of synaptic vesicles (SVs) at nerve terminals. The mechanisms involved in these processes are highly controversial with evidence being presented for SV membranes being retrieved exclusively via clathrin-mediated endocytosis (CME) from the plasma membrane or via ultrafast endocytosis independent of clathrin. Here we show that clathrin and its major adaptor protein 2 (AP-2) in addition to the plasma membrane operate at internal endosome-like vacuoles to regenerate SVs but are not essential for membrane retrieval. Depletion of clathrin or conditional knockout of AP-2 result in defects in SV reformation and an accumulation of endosome-like vacuoles generated by clathrin-independent endocytosis (CIE) via dynamin 1/3 and endophilin. These results together with theoretical modeling provide a conceptual framework for how synapses capitalize on clathrin-independent membrane retrieval and clathrin/AP-2-mediated SV reformation from endosome-like vacuoles to maintain excitability over a broad range of stimulation frequencies.
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