Synaptic vesicle (SV) endocytosis at the presynapse is essential for the maintenance of neurotransmission, particularly during intense neuronal activity. Under these conditions the dominant SV endocytosis mode is activity-dependent bulk endocytosis (ADBE), suggesting it should perform a central role in brain function. However determining the physiological role of ADBE in neurotransmission has been hindered by a limited understanding of its molecular mechanism. This project will address this challenge by discovering presynaptic molecules with a specific and selective role in ADBE. This will be achieved by exploiting the recently generated ADBE proteome – which has revealed a series of presynaptic molecules with no currently identified function. The role of these candidate molecules in ADBE and other endocytosis modes will be determined using a palette of live fluorescent imaging assays in primary neuronal cultures. Molecules that are specific to ADBE will to taken forward to determine their molecular mechanism using a series of biochemical interaction studies. In addition to this there is the opportunity to generate transgenic animals lacking specific molecules to determine their function in vivo. This project will therefore provide training in a series of state of the art technologies addressing a key biological problem.
Nicholson-Fish J.C., Kokotos, A.C., Gillingwater T.G., Smillie K.J.* and Cousin M.A.* (2015) VAMP4 is an essential cargo molecule for activity-dependent bulk endocytosis. Neuron 88: 973-984.
Smillie K.J., Pawson J, Perkins E.M., Jackson M. and Cousin M.A. (2013) Control of synaptic vesicle endocytosis by an extracellular signalling molecule. Nature Commun. 4: 2394.
Clayton E.L., Sue N., Smillie K.J., O’Leary T., Bache N., Cheung G., Cole, A.R., Wyllie D.J, Sutherland C., Robinson P.J. and Cousin M.A. (2010) Dynamin I phosphorylation by GSK3 controls activity-dependent bulk endocytosis of synaptic vesicles. Nature Neurosci. 13: 845-851.