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Updated: May 22, 2026

Measuring Synaptic Vesicle Endocytosis in Cultured Hippocampal Neurons
Published on: September 4, 2017
Adaptor protein complexes 1 and 3 are essential for generation of synaptic vesicles from activity-dependent bulk
Giselle Cheung1, Michael A Cousin
1Centre for Integrative Physiology, University of Edinburgh, Edinburgh, Scotland EH8 9XD, United Kingdom.
Abstract:
Activity-dependent bulk endocytosis is the dominant synaptic vesicle retrieval mode during high intensity stimulation in central nerve terminals. A key event in this endocytosis mode is the generation of new vesicles from bulk endosomes, which replenish the reserve vesicle pool. We have identified an essential requirement for both adaptor protein complexes 1 and 3 in this process by employing morphological and optical tracking of bulk endosome-derived synaptic vesicles in rat primary neuronal cultures. We show that brefeldin A inhibits synaptic vesicle generation from bulk endosomes and that both brefeldin A knockdown and shRNA knockdown of either adaptor protein 1 or 3 subunits inhibit reserve pool replenishment from bulk endosomes. Conversely, no plasma membrane function was found for adaptor protein 1 or 3 in either bulk endosome formation or clathrin-mediated endocytosis. Simultaneous knockdown of both adaptor proteins 1 and 3 indicated that they generated the same population of synaptic vesicles. Thus, adaptor protein complexes 1 and 3 play an essential dual role in generation of synaptic vesicles during activity-dependent bulk endocytosis.
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