The goal of this project is to advance knowledge of mechanisms in synaptic transmission. More specifically, the project will investigate pathways of synaptic vesicle endocytosis and recycling following the collapse of a fused synaptic vesicle into the plasma membrane. While it is well established that clathrin and dynamin- dependent endocytosis plays a major role in this process, key mechanistic aspects of this process remain elusive. In addition, there is evidence for clathrin and dynamin-independent bulk endocytic pathways at the synapse, but such pathways remain poorly understood. The role of endosomal compartments in the vesicle cycle also remains controversial. This grant application plans first to characterize endocytic traffic that operates in parallel to the classic clathrin-dependent endocytic pathway and to test the hypothesis that bulk endocytosis leads to the generation of new synaptic vesicles independently of clathrin and of the GTPase dynamin. Second, the project will investigate the properties, potential heterogeneity and interrelationships of endocytic compartments of nerve terminals, determine the function of 3-phosphorylated phosphoinositides in their dynamics and test the hypothesis that most intermediates in synaptic vesicle recycling are not classical endosomes. Third, mechanistic aspects of the clathrin and dynamin-dependent endocytic reaction will be investigated. These studies will involve mouse genetics, studies in cultured neurons and in cell-free systems. This work is of specific relevance in neuroscience, but given the fundamental nature of endocytosis and post- endosomal sorting, it will have broad implications for a variety of fields of biology and medicine.

Public Health Relevance

Synaptic vesicles are the secretory organelles that store and secrete neurotransmitters at neuronal synapses. It is well established that following their fusion with the plasma membrane during neurotransmitter release, synaptic vesicle membranes must be recycled for the generation of new synaptic vesicles, but the precise mechanisms underlying this process remain a matter of debate. The goal of this proposal is to capitalize on recent advances in imaging methods and on perturbations of synaptic vesicle recycling in genetically modified mice and in cells derived from such mice to advance understanding of this fundamental process that supports communication between neurons.

Agency
National Institute of Health (NIH)
Institute
National Institute of Neurological Disorders and Stroke (NINDS)
Type
Method to Extend Research in Time (MERIT) Award (R37)
Project #
5R37NS036251-20
Application #
9064220
Study Section
Special Emphasis Panel (ZRG1)
Program Officer
Talley, Edmund M
Project Start
1997-05-01
Project End
2017-04-30
Budget Start
2016-05-01
Budget End
2017-04-30
Support Year
20
Fiscal Year
2016
Total Cost
Indirect Cost
Name
Yale University
Department
Anatomy/Cell Biology
Type
Schools of Medicine
DUNS #
043207562
City
New Haven
State
CT
Country
United States
Zip Code
Dong, Rui; Zhu, Ting; Benedetti, Lorena et al. (2018) The inositol 5-phosphatase INPP5K participates in the fine control of ER organization. J Cell Biol 217:3577-3592
Kumar, Nikit; Leonzino, Marianna; Hancock-Cerutti, William et al. (2018) VPS13A and VPS13C are lipid transport proteins differentially localized at ER contact sites. J Cell Biol 217:3625-3639
Milovanovic, Dragomir; Wu, Yumei; Bian, Xin et al. (2018) A liquid phase of synapsin and lipid vesicles. Science 361:604-607
Benedetti, Lorena; Barentine, Andrew E S; Messa, Mirko et al. (2018) Light-activated protein interaction with high spatial subcellular confinement. Proc Natl Acad Sci U S A 115:E2238-E2245
Bian, Xin; Saheki, Yasunori; De Camilli, Pietro (2018) Ca2+ releases E-Syt1 autoinhibition to couple ER-plasma membrane tethering with lipid transport. EMBO J 37:219-234
Lees, Joshua A; Zhang, Yixiao; Oh, Michael S et al. (2017) Architecture of the human PI4KIII? lipid kinase complex. Proc Natl Acad Sci U S A 114:13720-13725
Gowrishankar, Swetha; Wu, Yumei; Ferguson, Shawn M (2017) Impaired JIP3-dependent axonal lysosome transport promotes amyloid plaque pathology. J Cell Biol 216:3291-3305
Ritter, Brigitte; Ferguson, Shawn M; De Camilli, Pietro et al. (2017) A lentiviral system for efficient knockdown of proteins in neuronal cultures [version 1; referees: 2 approved]. MNI Open Res 1:
Saheki, Yasunori; De Camilli, Pietro (2017) Endoplasmic Reticulum-Plasma Membrane Contact Sites. Annu Rev Biochem 86:659-684
Cao, Mian; Wu, Yumei; Ashrafi, Ghazaleh et al. (2017) Parkinson Sac Domain Mutation in Synaptojanin 1 Impairs Clathrin Uncoating at Synapses and Triggers Dystrophic Changes in Dopaminergic Axons. Neuron 93:882-896.e5

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