Tomographie Elektronen-Mikroskop
Zusammenfassung der Projektergebnisse
The TEM is used by a number of groups located in different institutes of the University of Göttingen, in the faculties of Biology, Geology, Physics and Medicine. It is also used by collaboration partners from non-university institutes in Göttingen. Finally, several users collaborate with researchers from other sites in Germany and abroad. We indicate here only the most important findings, of the main users. Rizzoli group (Medicine Faculty). The TEM (JEM-1011) was used to generate threedimensional views of synaptic boutons. The project involved the generation of a map of an average synaptic bouton, in molecular detail, based on a combination of electron microscopy, biochemistry, super-resolution light microscopy and modeling. The TEM was crucial in this process, enabling the group to show with precision the shape, volume and number of synaptic elements. In addition, vesicle trafficking in sensory cells was characterized by using fluorescence photo-oxidation electron microscopy. Here the technique of transforming fluorescent probes into electron microscopy labels was applied for the first time to a sensory system. Moser group (Medicine Faculty). The JEM-1011 was especially useful in the determination of the morphology of membranes participating in synaptic vesicle recycling in sensory cells of the auditory pathway (in the inner hair cells). The organization of the active zone, where synaptic vesicles fuse with the plasma membrane to release their neurotransmitter contents, was analyzed as well. These works were performed in collaboration with the Wichmann group. Wichmann group (Medicine Faculty). The group published several articles in collaboration with the Moser group (see previous paragraph). The group also collaborated with the group of Stephan Sigrist (Free University, Berlin), in investigating the roles of mutants of the active zone protein Bruchpilot in the Drosophila neuromuscular junction. This protein is central for the formation of active zones in this preparation. The work demonstrates that several different isoforms participate in the active zone formation. Hoppert group (Biology Faculty). The group uses transmission electron microscopy to study prokaryotes in microbial biofilms, to investigate the formation of biominerals, and to study the relation between microbial symbionts and their hosts (such as molluscs, crustaceans or sponges). Eimer group (European Neuroscience Institute, Göttingen). The Eimer group used the TEM to study the involvement of a novel protein, TFG-1, in the export of cargoes from the endoplasmic reticulum (ER). The work was performed in Caenorhabditis elegans, and it demonstrates the role of this molecule in protein exit from the ER. Varoqueaux and Brose groups (Max Planck Institute for Experimental Medicine, Göttingen). They studied the organization of synapses in the mouse retina, focusing on the involvement of the protein Munc-13. Electron tomography was used to determine whether Munc-13 is involved in vesicle priming. Sigrist group (Free University, Berlin). The group used the JEM-2100 device, in collaboration with C. Wichmann, to investigate the involvement of the protein RBP (RIM-binding protein) in the formation and function of the Drosophila active zone. This was the first demonstration of the involvement of RBP in the generation of the so-called T-bar at the Drosophila active zone.
Projektbezogene Publikationen (Auswahl)
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TFG-1 function in protein secretion and oncogenesis. Nat Cell Biol. 2011 May;13(5):550-8
Witte K1, Schuh AL, Hegermann J, Sarkeshik A, Mayers JR, Schwarze K, Yates JR 3rd, Eimer S, Audhya A
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(2012). Munc13-independent vesicle priming at mouse photoreceptor ribbon synapses. J Neurosci. 2012 June 6;32(23):8040-52
Cooper B, Hemmerlein M, Ammermüller J, Imig C, Reim K, Lipstein N, Kalla S, Kawabe H, Brose N, Brandstätter JH, Varoqueaux F
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(2013). Box RNA helicases in Bacillus subtilis have multiple functions and act independently from each other. J Bacteriol 195, 534-544
Lehnik-Habrink M, Rempeters L, Kovács ÁT, Wrede C, Baierlein C, Krebber H, Kuipers OP, Stülke J
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(2013). Localization of methyl-coenzyme M reductase as metabolic marker for diverse methanogenic Archaea. Archaea 2013, ID 920241
Wrede C, Walbaum U, Ducki A, Heieren I, Hoppert M
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(2013). The Bruchpilot cytomatrix regulates the readilyreleasable pool of synaptic vesicles. JCB 202, 667-83
Matkovic T, Siebert M, Knoche E, Depner H, Mertel S, David Owald, Schmidt M, Thomas U, Sickmann A, Kamin D, Hell SW, Bürger J, Hollmann C, Mielke T, Wichmann C, Sigrist SJ
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(2011). RIM-Binding Protein, a Central Part of the Active Zone, Is Essential for Neurotransmitter Release. Science 2014 334, 1565-1569
Liu KSY, Siebert M, Mertel S, Knoche E, Wegener S, Wichmann C, Matkovic T, Muhammad K, Depner H, Mettke C, Bückers J, Hell SW, Müller M, Davis GW, Schmitz D, Sigrist SJ
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(2014) Characterization and phylogeny of a novel methanotroph, Methyloglobulus morosus gen. nov., spec. nov. Syst Appl Microbiol 37, 165-169
Deutzmann JS, Hoppert M, Schink B
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(2014). Developmental refinement of hair cell synapses tightens the coupling of Ca2+ influx to exocytosis. EMBO 33, 247-64
Wong AB, Rutherford MA, Gabrielaitis M, Pangršič T, Göttfert F, Frank T, Michanski S, Hell S, Wolf F, Carolin Wichmann C, Moser T
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(2014). Modes and regulation of endocytic membrane retrieval in mouse auditory hair cells. J Neurosci 34, 705-16
Neef J, Jung SY, Wong AB, Reuter K, Pangršič T, Chakrabarti R, Kügler S, Lenz C, Nouvian R, RM Boumil, Frankel WN, Wichmann C, Moser T
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Bassoon-disruption slows vesicle replenishment and induces homeostatic plasticity at a CNS synapse. EMBO J. 2014 Mar 3;33(5):512-27
Mendoza-Schulz A, Jing Z, Sánchez Caro JM, Wetzel F, Dresbach T, Strenzke N, Wichmann C, Moser T
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Composition of isolated synaptic boutons reveals the amounts of vesicle trafficking proteins. Science. 2014 May 30;344(6187):1023-8
Wilhelm BG, Mandad S, Truckenbrodt S, Kröhnert K, Schäfer C, Rammner B, Koo SJ, Claßen GA, Krauss M, Haucke V, Urlaub H, Rizzoli SO
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FM dye photo-oxidation as a ool for monitoring membrane recycling in inner hair cells. PLoS One. 2014 Feb 5;9(2):e88353
Kamin D, Revelo NH, Rizzoli SO
