STED Konfokalmikroskop

• Subject Area: Basic Research in Biology and Medicine
• Term: Funded in 2008
• Project identifier: Deutsche Forschungsgemeinschaft (DFG) - Project number 70427742

• Final Report Year: 2012

Final Report Abstract

AG Stephan Sigrist (Fouquet, Andlauer): The majority of rapid cell-to-cell communication mechanisms and information processing within the nervous system makes use of chemical synapses. Thereby, the molecular organization of presynaptic active zones, the places where neurotransmitter filled synaptic vesicles get released, is a focus of intense investigation. We recently showed that Bruchpilot (BRP) of Drosophila melanogaster, which features homologies to the mammalian CAST/ERC family, is essential for structural organization and efficient neurotransmitter release at active zones. The light microscopic inspection of synaptic organization is often restricted by the limited resolution of conventional light microcopy due to diffraction. Thus, we stimulated emission depletion microscopy, STED for the analysis of synapse substructures. STED breaks the diffraction barrier and allows localization of proteins well below 100 nm. Using STED, we showed that BRP shapes the presynaptic active zone architecture by adopting an extended conformation, shining first light on the underlying macromolecular organization of active zones. AG Nieswandt (Gupta): STED analysis of the platelet cytoskeleton using different transgenic mouse models to study dynamics, subcellular localization of different signaling molecules and actin cytoskeleton in spreading platelets. AG Harms (Friedrich): Here, STED was combined with selective plane illumination microscopy (SPIM). Using this method, an up to 60% improvements in axial resolution with lateral resolution enhancements in control samples and zebrafish embryos coudl be reached. The integrated STED-SPIM method combines the advantages of SPIM with the resolution enhancement of STED, and thus provides a method for fast, high-resolution imaging with >100 µm deep penetration into biological tissue.

Publications

• Maturation of active zone assembly by Drosophila Bruchpilot. J Cell Biol 2009, 186(1):129-45
  W Fouquet, D Owald, C Wichmann, S Mertel, H Depner, M Dyba, S Hallermann, RJ Kittel, S Eimer, SJ Sigrist
  
• A Syd-1 homologue regulates pre- and postsynaptic maturation in Drosophila J Cell Biol 2010, 188(4):565-79
  D Owald, W Fouquet, M Schmidt, C Wichmann, S Mertel, H Depner, F Christiansen, C Zube, C Quentin, J Körner, H Urlaub, K Mechtler, SJ Sigrist
  
• ADF/n-cofilin-dependent actin turnover determines platelet formation and sizing. Blood 2010, 116(10):1767-75
  M Bender, A Eckly, JH Hartwig, M Elvers, I Pleines, S Gupta, G Krohne, E Jeanclos, A Gohla, C Gurniak, C Gachet, W Witke, B Nieswandt
  
• Naked Dense Bodies Provoke Depression. J Neuroscience 2010, 30(43):14340-45
  S Hallermann, RJ Kittel, C Wichmann, A Weyhersmüller, W Fouquet, S Mertel, D Owald, S Eimer, H Depner, M Schwärzel, SJ Sigrist, M Heckmann
  
• Stiff person syndrome-associated autoantibodies to amphiphysin mediate reduced GABAergic inhibition. Brain 2010, 133(11):3166-80
  C Geis, A Weishaupt, S Hallermann, B Grünewald, C Wessig, T Wultsch, A Reif, N Byts, M Beck, S Jablonka, MK Boettger, N Uçeyler, W Fouquet, M Gerlach, HM Meinck, AL Sirén, SJ Sigrist, KV Toyka, M Heckmann, C Sommer
  
• Piccolo Regulates the Dynamic Assembly of Presynaptic F-Actin. J Neuroscience 2011, 31(40):14250-63
  CL Waites, SA Leal-Ortiz, TFM Andlauer, SJ Sigrist,CC Garner
  
• Rapid active zone remodeling during synaptic plasticity. J Neuroscience 2011, 31(16):6041-52
  A Weyhersmüller, S Hallermann, N Wagner, J Eilers
  
• STED-SPIM: Stimulated emission depletion improves sheet illumination microscopy resolution. Biophys J 2011, 100(8):L43-45
  M Friedrich, Q Gan, V Ermolayev, GS Harms
  
• Megakaryocyte-specific RhoA deficiency causes macrothrombocytopenia and defective platelet activation in hemostasis and thrombosis. Blood 2012, 119(4):1054-63
  I Pleines, I Hagedorn, S Gupta, F May, L Chakarova, J van Hengel, S Offermanns, G Krohne, C Kleinschnitz, C Brakebusch, B Nieswandt
  
• Role of Nav1.9 in activity-dependent axon growth in motoneurons. Hum Mol Genet 2012, 21(16):3655-67
  N Subramanian, A Wetzel, B Dombert, P Yadav, S Havlicek, S Jablonka, MA Nassar, R Blum, M Sendtner