Objective Stimulated Emission Depletion (STED) microscopy is one of the most important recent developments in light microscopy (Willig et al., 2006, Nature 440:935-9). STED allows for imaging cellular elements with diffraction-unlimited resolution; in practical terms, the resolution (normally limited to ~200-300 nm) is improved down to 30-60 nm. Together with the development of two-color STED microscopy (Donnert et al., 2007, Biophys J. 92:L67-9), this technique allows experimenters to pinpoint the position of various cellular elements with nanometer precision. Obtaining a cellular nanomap is not feasible with conventional light microscopy, due to its low resolution. Electron microscopy cannot be applied, as its labeling efficiency it too low. I propose here to use STED microscopy to characterize the positions of the major components of the synapse. The preparation will be cultured hippocampal neurons, which have numerous small (about one micron in diameter) synaptic nerve terminals. I will determine the locations of synaptic proteins involved in neurotransmitter release, in membrane retrieval and in pre- and post-synaptic active zone structure. Less specialized elements such as the cytoskeleton, mitochondria and endosomes of the synapse will also be investigated. The work will provide answers for a number of questions in the neuroscience field, such as how and where the synaptic vesicles get retrieved, how pre- and post-synaptic active zone elements correlate, and what the role of cytoskeletal elements is in synaptic transmission. The small size and relatively low complexity (compared to whole cells) of the synaptic boutons will allows the work to be completed within a reasonable timeframe. Successful completion of the project will encourage researchers to perform larger scale cellular nano-maps, which would eventually replace the largely erroneous cellular fractionation techniques currently used nowadays to determine the location of various proteins. Fields of science natural sciencesbiological sciencesneurobiologynatural sciencesbiological sciencesbiochemistrybiomoleculesproteinsnatural sciencesphysical sciencesopticsmicroscopyelectron microscopy Keywords (STED) Depletion Emission Microscopy Resolution Stimulated Stimulated Emission Depletion (STED) Super Programme(s) FP7-IDEAS-ERC - Specific programme: "Ideas" implementing the Seventh Framework Programme of the European Community for research, technological development and demonstration activities (2007 to 2013) Topic(s) ERC-SG-LS4 - ERC Starting Grant - Physiology, Pathophysiology and Endocrinology Call for proposal ERC-2007-StG See other projects for this call Funding Scheme ERC-SG - ERC Starting Grant Coordinator UNIVERSITAETSMEDIZIN GOETTINGEN - GEORG-AUGUST-UNIVERSITAET GOETTINGEN - STIFTUNG OEFFENTLICHEN RECHTS Address Robert-koch-strasse 40 37075 Goettingen Germany See on map Region Niedersachsen Braunschweig Göttingen Activity type Higher or Secondary Education Establishments Administrative Contact Christiane Hennecke (Ms.) Principal investigator Silvio Olivier Rizzoli (Dr.) Links Contact the organisation Opens in new window Website Opens in new window EU contribution € 1 670 000,00 Beneficiaries (1) Sort alphabetically Sort by EU Contribution Expand all Collapse all UNIVERSITAETSMEDIZIN GOETTINGEN - GEORG-AUGUST-UNIVERSITAET GOETTINGEN - STIFTUNG OEFFENTLICHEN RECHTS Germany EU contribution € 1 670 000,00 Address Robert-koch-strasse 40 37075 Goettingen See on map Region Niedersachsen Braunschweig Göttingen Activity type Higher or Secondary Education Establishments Administrative Contact Christiane Hennecke (Ms.) Principal investigator Silvio Olivier Rizzoli (Dr.) Links Contact the organisation Opens in new window Website Opens in new window
