TRR 166:  High-end light microscopy elucidates membrane receptor function - ReceptorLight

• Subject Area: Biology ; Chemistry ; Medicine ; Physics
• Term: from 2015 to 2020
• Project identifier: Deutsche Forschungsgemeinschaft (DFG) - Project number 258780946

The total of processes enabling life of a cell is unimaginable without inclusion of functional membrane receptors. Besides the role of receptors in keeping the homeostasis of life, they also gained outstanding relevance for pharmacology and curative medicine. Technologically, in the past decade enormous progress has been made in high-end light microscopic techniques. Combining such techniques with recent relevant developments in the fields of fluorescent dyes and probes as well as adequate functional assays has become exceptionally powerful on the way to reach a new level of understanding the function of membrane receptors.In an overarching strategy, ranging from technical research in microscopy over biophysics, receptor physiology and pharmacology to application in medicine, the TRR 166 ReceptorLight has three main goals:1) Receptor localization will be studied at the highest possible resolution in space and time using all presently available methods of super-resolution microscopy, thereby developing novel optical techniques, refined data analyses, fluorescent dyes, probes and optogenetic tools.2) Novel insight into the function of both metabotropic and ionotropic receptors will be gained by high-end light microscopic techniques on elementary processes such as ligand binding, conformational changes of the binding domains and subunits, cooperative interactions of the subunits, and conformational changes of whole receptors as well as their interaction with other proteins. Based on known structures, MD simulations in combination with mutagenesis will be employed to understand the dynamics of selected receptors. 3) High-end light-microscopic strategies will be developed and used to improve the understanding of the role of receptors in selected diseases. It is expected that these studies become paradigms for how high-end microscopic approaches can be used advantageously in diagnosis and the development of more efficient and safer therapies.Overall, the operating principle in the TRR ReceptorLight is iterative, new results on receptor localization and function will drive the generation of new techniques which in turn will generate new results on the receptors and so on.

• DFG Programme: CRC/Transregios

Completed projects

• A01 - Black-lipid-membrane platform with diffusion suppression for confocal single-molecule FLIM-FRET analysis of gated membrane receptors and transporters (Project Head Börsch, Michael )
• A02 - Label-free investigation of structure-dynamic relations of single membrane receptors using a tip-enhanced Raman scattering approach (Project Head Deckert, Volker )
• A03 - Optogenetic control of ionotropic receptor function by cyclic nucleotides (Project Heads Kittel, Robert J. ;  Nagel, Georg )
• A04 - Genetic code expansion and bioorthogonal click labeling for super-resolution imaging of receptors with molecular resolution (Project Heads Heintzmann, Rainer ;  Kollmannsberger, Philip ;  Sauer, Markus )
• A05 - Relating the binding of single ligands to activation in single HCN and CNG channels (Project Heads Benndorf, Klaus ;  Schmauder, Ralf )
• A09 - Deciphering fast intra- and intermolecular dynamics of G protein-coupled receptors by Fluorescence (Cross) Correlation Spectroscopy (Project Heads Heinze, Katrin G. ;  Lohse, Martin J. )
• B01 - Elucidating assembly, ligand binding and gating of heterotetrameric CNG channels by FRET (Project Heads Biskup, Christoph ;  Nache, Vasilica )
• B03 - Interaction of GABAA-receptor function and network activity in the developing hippocampus (Project Heads Holthoff, Knut ;  Kirmse, Knut )
• B04 - Activity-induced ionotropic glutamate receptor dynamics at super-resolution in vivo (Project Heads Kittel, Robert J. ;  Sauer, Markus )
• B05 - Enlightening inhibitory neurotransmitter receptor organization and dynamics applying high end microscopy (Project Heads Heintzmann, Rainer ;  Maric, Hans Michael ;  Qualmann, Britta )
• B06 - Functional plasticity of glutamate receptor channels on hippocampal mossy fiber terminals (Project Heads Heckmann, Manfred ;  Sirén, Anna-Leena K. )
• B07 - Relating ligand binding and activation gating in nicotinic acetylcholine receptors (Project Head Kusch, Jana )
• B08 - Plasticity and dynamic interactions of chitin-induced receptor-mediated innate immunity (Project Heads Geiger, Dietmar ;  Hedrich, Rainer )
• C01 - Spatial organization and dynamics of GPCR signaling as revealed by single-molecule and superresolution microscopy (Project Head Calebiro, Davide )
• C02 - The interplay of receptor dynamics and ligand residence time upon ligand efficacy at adenosine receptors (Project Head Hoffmann, Carsten )
• C03 - FRET-based monitoring of Adhesion class G protein-coupled receptor activity (Project Head Langenhan, Tobias )
• C04 - Kinetics of metabotropic glutamate receptors (Project Heads Benndorf, Klaus ;  Lohse, Martin J. )
• C05 - Real-time imaging of μ-opioid receptor signaling, phosphorylation and desensitization (Project Head Schulz, Stefan )
• C07 - Signaling mechanism, subcellular localization and regulation of guanylyl cyclases A and B, the receptors for atrial and C-type natriuretic peptides (Project Head Kuhn, Michaela )
• D02 - Investigating humoral autoimmunity against the NMDA-receptor NR1 subunit by super-resolution fluorescence microscopy: effects on synapse integrity and function (Project Heads Doose, Sören ;  Geis, Christian )
• Z02 - Integrative data management and processing (Project Heads Bücker, H. Martin ;  Henkel, Andreas G. ;  König-Ries, Birgitta )
• Z03 - Central administration and project coordination (Project Head Benndorf, Klaus )

• Applicant Institution: Friedrich-Schiller-Universität Jena
• Co-Applicant Institution: Julius-Maximilians-Universität Würzburg
• Participating Institution: Leibniz-Institut für Photonische Technologien e.V. (IPHT)
• Participating University: Heinrich-Heine-Universität Düsseldorf; Universität Leipzig
• Spokesperson: Professor Dr. Klaus Benndorf