Synaptic vesicle cycle at nano-resolution

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Publication Type dissertation
School or College College of Science
Department Biology
Author Watanabe, Shigeki
Title Synaptic vesicle cycle at nano-resolution
Date 2013-05
Description Electron microscopy can visualize synapses at nanometer resolution, and can thereby capture the fine structure of these contacts. However, this imaging method lacks three key elements: temporal information, protein visualization, and large volume reconstruction. For my dissertation, I developed three methods in electron microscopy that overcame these limitations. First, I developed a method to freeze neurons at any desired time point after a stimulus to study synaptic vesicle cycle. Second, I developed a method to couple super-resolution fluorescence microscopy and electron microscopy to pinpoint the location of proteins in electron micrographs at nanometer resolution. Third, I collaborated with computer scientists to develop methods for semi-automated reconstruction of nervous system. I applied these techniques to answer two fundamental questions in synaptic biology. Which vesicles fuse in response to a stimulus? How are synaptic vesicles recovered at synapses after fusion? Only vesicles that are in direct contact with plasma membrane fuse upon stimulation. The active zone in C. elegans is broad, but primed vesicles are concentrated around the dense projection. Following exocytosis of synaptic vesicles, synaptic vesicle membrane was recovered rapidly at two distinct locations at a synapse: the dense projection and adherens junctions. These studies suggest that there may be a novel form of ultrafast endocytosis.
Type Text
Publisher University of Utah
Subject advanced microscopy methods; correlative microscopy; neurotransmission; super-resolution fluorescence microscopy; synaptic vesicle cycle; time-resolved electron microscopy
Dissertation Name Doctor of Philosophy
Language eng
Rights Management Copyright © Shigeki Watanabe 2013
Format Medium application/pdf
Format Extent 7,097,113 bytes
ARK ark:/87278/s6sn0qtn
Setname ir_etd
Date Created 2013-05-17
Date Modified 2018-04-17
ID 195892
Reference URL