Control of synaptobrevin retrieval by synaptophysin

Synaptobrevin-2 and synaptophysin are the two most abundant proteins present on a synaptic vesicle (SV). Synaptobrevin-2 is a protein essential for SV exocytosis, whereas until recently synaptophysin was a protein of unknown function. We have recently shown that synaptophysin is essential for the specific retrieval of synaptobrevin-2 from the plasma membrane after exocytosis, however may questions remain unanswered. These include, what controls the retrieval of synaptophysin itself, how does synaptophysin interact with synaptobrevin-2, and does synaptophysin also control the retrieval of non-canonical synaptobrevins (thought to define different modes of exocytosis)? Approaches used will include primary neuronal cultures derived from synaptophysin knockout mice, monitoring of SV recycling using fluorescent genetic and chemical reporters, ultrastructural analysis using transmission electron microscopy, recombinant protein expression, classical biochemistry and molecular biology technologies.