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

• Final Report Year: 2021

Final Report Abstract

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 applications in medicine, the TRR 166 ReceptorLight focused its research on three main goals: 1) Receptor localization was 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 insights into the function of both metabotropic and ionotropic receptors were 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 interactions with other proteins. 3) High-end light-microscopic strategies were 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 was iterative, new results on receptor localization and function pushed the generation of new techniques, which in turn were and will also in the future be used advantageously to study membrane receptors distribution, function, dynamics, signaling, and infection pathways, including infections with corona viruses.

Publications

• (2016) Adhesion G protein-coupled receptors in nervous system development and disease. Nat. Rev. Neurosci. 17, 550
  Langenhan, T., Piao, X., and Monk, K.R.
  (See online at https://doi.org/10.1038/nrn.2016.86)
• (2016) cAMP signals in Drosophila motor neurons are confined to single synaptic boutons. Cell Rep. 17, 1238-1246
  Maiellaro, I., Lohse, M.J., Kittel, R.J., and Calebiro, D.
  (See online at https://doi.org/10.1016/j.celrep.2016.09.090)
• (2016) Deciphering the function of the CNGB1b subunit in olfactory CNG channels. Sci. Rep. 6, 29378
  Nache, V., Wongsamitkul, N., Kusch, J., Zimmer, T., Schwede, F., and Benndorf, K.
  (See online at https://doi.org/10.1038/srep29378)
• (2016) Gephyrin super-binding peptides visualize inhibitory synapses and modulate inhibitory transmission. Nat. Chem. Biol. 13, 153
  Maric, H.M., Dalby, N.O., Hausrat, T.J., Neubert, F., Doose, S., Sauer, M., Kneussel, M., and Strømgaard, K.
  (See online at https://doi.org/10.1038/nchembio.2246)
• (2016) High resolution spectroscopy reveals fibrillation inhibition pathways of insulin. Sci. Rep. 6, 39622
  Deckert-Gaudig, T., and Deckert, V.
  (See online at https://doi.org/10.1038/srep39622)
• (2016) Quantifying the cooperative subunit action in a multimeric membrane receptor. Sci. Rep. 6, 20974
  Wongsamitkul, N., Nache, V., Eick, T., Hummert, S., Schulz, E., Schmauder, R., Schirmeyer, J., Zimmer, T., and Benndorf, K.
  (See online at https://doi.org/10.1038/srep20974)
• (2016) Silent S-Type Anion Channel Subunit SLAH1 Gates SLAH3 Open for Chloride Root-to-Shoot Translocation. Curr. Biol. 26, 2213-2220
  Cubero-Font, P., Maierhofer, T., Jaslan, J., Rosales, M.A., Espartero, J., Díaz-Rueda, P., Müller, H.M., Hürter, A.L., Al-Rasheid, K.A.S., Marten, I., Hedrich, R., Colmenero-Flores, J.M., and Geiger, D.
  (See online at https://doi.org/10.1016/j.cub.2016.06.045)
• (2016) β-Arrestin biosensors reveal a rapid, receptor-dependent activation/deactivation cycle. Nature 531, 661-664
  Nuber, S., Zabel, U., Lorenz, K., Nuber, A., Milligan, G., Tobin, A.B., Lohse, M.J., and Hoffmann, C.
  (See online at https://doi.org/10.1038/nature17198)
• (2017) Autoantibodies to Synaptic Receptors and Neuronal Cell Surface Proteins in Autoimmune Diseases of the Central Nervous System. Physiol. Rev. 97, 839–887
  Dalmau, J., Geis, C., and Graus, F.
  (See online at https://doi.org/10.1152/physrev.00010.2016)
• (2017) Defective synaptic transmission causes disease signs in a mouse model of juvenile neuronal ceroid lipofuscinosis. eLife 6, e28685
  Grünewald, B., Lange, M.D., Werner, C., O'Leary, A., Weishaupt, A., Popp, S., Pearce, D.A., Wiendl, H., Reif, A., Pape, H.C., Toyka, K.V., Sommer, C., and Geis, C.
  (See online at https://doi.org/10.7554/elife.28685)
• (2017) Emerging Paradigms of G Protein-Coupled Receptor Dephosphorylation. Trends Pharmacol. Sci. 38, 621-636
  Kliewer, A., Reinscheid, R.K., and Schulz, S.
  (See online at https://doi.org/10.1016/j.tips.2017.04.002)
• (2017) Evolutionary action and structural basis of the allosteric switch controlling β2AR functional selectivity. Nat. Commun. 8, 2169
  Schönegge, A.M., Gallion, J., Picard, L.P., Wilkins, A.D., Le Gouill, C., Audet, M., Stallaert, W., Lohse, M.J., Kimmel, M., Lichtarge, O., and Bouvier, M.
  (See online at https://doi.org/10.1038/s41467-017-02257-x)
• (2017) Internalized TSH receptors en route to the TGN induce local Gs-protein signaling and gene transcription. Nat. Commun. 8, 443
  Godbole, A., Lyga, S., Lohse, M.J., and Calebiro, D.
  (See online at https://doi.org/10.1038/s41467-017-00357-2)
• (2017) Mechano-dependent signaling by Latrophilin/CIRL quenches cAMP in proprioceptive neurons. eLife 6, e28360
  Scholz, N., Guan, C., Nieberler, M., Grotemeyer, A., Maiellaro, I., Gao, S., Beck, S., Pawlak, M., Sauer, M., Asan, E., Rothemund, S., Winkler, J., Prömel, S., Nagel, G., Langenhan, T., and Kittel, R.J.
  (See online at https://doi.org/10.7554/elife.28360)
• (2017) Single-molecule imaging reveals receptor–G protein interactions at cell surface hot spots. Nature 550, 543
  Sungkaworn, T., Jobin, M.L., Burnecki, K., Weron, A., Lohse, M.J., and Calebiro, D.
  (See online at https://doi.org/10.1038/nature24264)
• (2018) A Tandem Amino Acid Residue Motif in Guard Cell SLAC1 Anion Channel of Grasses Allows for the Control of Stomatal Aperture by Nitrate. Curr. Biol. 28, 1370-1379
  Schäfer, N., Maierhofer, T., Herrmann, J., Jorgensen, M.E., Lind, C., von Meyer, K., Lautner, S., Fromm, J., Felder, M., Hetherington, A.M., Ache, P., Geiger, D., and Hedrich, R.
  (See online at https://doi.org/10.1016/j.cub.2018.03.027)
• (2018) Activation gating in HCN2 channels. PLoS Comput. Biol. 14, e1006045
  Hummert, S., Thon, S., Eick, T., Schmauder, R., Schulz, E., and Benndorf, K.
  (See online at https://doi.org/10.1371/journal.pcbi.1006045)
• (2018) Bioorthogonal click chemistry enables site-specific fluorescence labeling of functional NMDA receptors for super-resolution imaging. Angew. Chem. Int. Ed. 57, 16364-16369
  Neubert, F., Beliu, G., Terpitz, U., Werner, C., Geis, C., Sauer, M., and Doose, S.
  (See online at https://doi.org/10.1002/anie.201808951)
• (2018) Endothelial C-Type Natriuretic Peptide Acts on Pericytes to Regulate Microcirculatory Flow and Blood Pressure. Circulation 138, 494
  Špiranec, K., Chen, W., Werner, F., Nikolaev, V.O., Naruke, T., Koch, F., Werner, A., Eder-Negrin, P., Diéguez-Hurtado, R., Adams, R.H., Baba, H.A., Schmidt, H., Schuh, K., Skryabin, B.V., Movahedi, K., Schweda, F., and Kuhn, M.
  (See online at https://doi.org/10.1161/circulationaha.117.033383)
• (2018) Enhanced fluorescence resonance energy transfer in G protein-coupled receptor probes by nano-coated microscopy coverslips. ACS Photonics 5, 2225-2233
  Schreiber, B., Kauk, M., Heil, H.S., Emmerling, M., Tessmer, I., Kamp, M. Höfling, S., Holzgrabe, U., Hoffmann, C., and Heinze, K.G.
  (See online at https://doi.org/10.1021/acsphotonics.8b00072)
• (2018) FZD5 is a Gαq-coupled receptor exhibiting the functional hallmarks of prototypical GPCRs. Sci. Signal. 11, eaar5536
  Wright, S.C., Alonso Canizal, M.C., Benkel, T., Simon, K., Le Gouill, C., Matricon, P., Namkung, Y., Lukasheva, V., König, G.M., Laporte, S.A., Carlsson, J., Kostenis, E., Bouvier, M., Schulte, G., and Hoffmann, C.
  (See online at https://doi.org/10.1126/scisignal.aar5536)
• (2018) Human autoantibodies against the AMPA receptor subunit GluA2 induce receptor reorganisation and memory dysfunction. Neuron 100, 91-105
  Haselmann, H., Mannara, F., Werner, C., Planaguma, J., Miguez-Cabalo, F., Schmidl, L., Grünewald, B., Petit-Pedrol, M., Kirmse, K., Classen, J., Demir, F., Klöcker, N., Soto, D., Doose, S., Dalmau, J., Hallermann, S., and Geis, C.
  (See online at https://doi.org/10.1016/j.neuron.2018.07.048)
• (2018) Hydrophobic alkyl chains substituted to the 8-position of cyclic nucleotides enhance activation of CNG and HCN channels by an intricate enthalpy - entropy compensation. Sci. Rep. 8, 14960
  Otte, M., Schweinitz, A., Bonus, M., Enke, U., Schumann, C., Gohlke, H., and Benndorf, K.
  (See online at https://doi.org/10.1038/s41598-018-33050-5)
• (2018) Multisite phosphorylation is required for sustained interaction with GRKs and arrestins during rapid mu-opioid receptor desensitization. Sci. Signal. 11, aas9609
  Miess, E., Gondin, A.B., Yousuf, A., Steinborn, R., Mosslein, N., Yang, Y., Goldner, M., Ruland, J.G., Bunemann, M., Krasel, C., Christie, M.J., Halls, M.L., Schulz, S., and Canals, M.
  (See online at https://doi.org/10.1126/scisignal.aas9609)
• (2019) Agonist-selective NOP receptor phosphorylation correlates in vitro and in vivo and reveals differential post-activation signaling by chemically diverse agonists. Sci. Signal. 12, eaau8072
  Mann, A., Moulédous, L., Froment, C., O'Neill, P.R., Dasgupta, P., Günther, T., Brunori, G., Kieffer, B.L., Toll, L., Bruchas, M.R., Zaveri, N.T., and Schulz, S.
  (See online at https://doi.org/10.1126/scisignal.aau8072)
• (2019) Anion channel SLAH3 is a regulatory target of chitin receptor-associated kinase PBL27 in microbial stomatal closure. eLife 8, e44474
  Liu, Y., Maierhofer, T., Rybak, K., Sklenar, J., Breakspear, A., Johnston, M.G., Fliegmann, J., Huang, S., Roelfsema, M.R.G., Felix, G., Faulkner, C., Menke, F.L., Geiger, D., Hedrich, R., and Robatzek, S.
  (See online at https://doi.org/10.7554/elife.44474)
• (2019) Bioorthogonal labeling with tetrazine-dyes for super-resolution microscopy. Commun. Biol. 2, 261
  Beliu, G., Kurz, A.J., Kuhlemann, A.C., Behringer-Pliess, L., Meub, M., Wolf, N., Seibel, J., Shi, Z.D., Schnermann, M., Grimm, J.B., Lavis, L.D., Doose, S., and Sauer, M.
  (See online at https://doi.org/10.1038/s42003-019-0518-z)
• (2019) Detection of Chlamydia Developmental Forms and Secreted Effectors by Expansion Microscopy. Front. Cell. Infect. Microbiol. 9, 276
  Kunz, T.C., Götz, R., Sauer, M., and Rudel, T.
  (See online at https://doi.org/10.3389/fcimb.2019.00276)
• (2019) Helix 8 is the essential structural motif of mechanosensitive GPCRs. Nat. Commun. 10, 5784
  Erdogmus, S., Storch, U., Danner, L., Becker, J., Winter, M., Ziegler, N., Wirth, A., Offermanns, S., Hoffmann, C., Gudermann, T., and Schnitzler, M.
  (See online at https://doi.org/10.1038/s41467-019-13722-0)
• (2019) Phosphorylation-deficient G-protein-biased μ-opioid receptors improve analgesia and diminish tolerance but worsen opioid side effects. Nat. Commun. 10, 367
  Kliewer, A., Schmiedel, F., Sianati, S., Bailey, A., Bateman, J.T., Levitt, E.S., Williams, J.T., Christie, M.J., and Schulz, S.
  (See online at https://doi.org/10.1038/s41467-018-08162-1)
• (2019) Stepwise activation of a class C GPCR begins with millisecond dimer rearrangement. Proc. Natl. Acad. Sci. U S A 116, 10150-10155
  Grushevskyi, E.O., Kukaj, T., Schmauder, R., Bock, A., Zabel, U., Schwabe, T., Benndorf, K., and Lohse, M.J.
  (See online at https://doi.org/10.1073/pnas.1900261116)
• (2019) Video-rate multi-color structured illumination microscopy with simultaneous real-time reconstruction. Nat. Commun. 10, 4315
  Markwirth, A., Lachetta, M., Mönkemöller, V., Heintzmann, R., Hübner, W., Huser, T., Müller, M.
  (See online at https://doi.org/10.1038/s41467-019-12165-x)
• (2020) G Protein-Coupled Receptor Pharmacology at the Single-Molecule Level. Annu. Rev. Pharmacol. Toxicol. 60, 73-87
  Calebiro, D., and Grimes, J.
  (See online at https://doi.org/10.1146/annurev-pharmtox-010919-023348)
• (2020) Single-molecule analysis reveals agonist-specific dimer formation of µ-opioid receptors. Nat. Chem. Biol. 16, 946-954
  Möller, J., Isbilir, A., Sungkaworn, T., Osberg, B., Karathanasis, C., Sunkara, V., Grushevskyi, E.O., Bock, A., Annibale, P., Heilemann, M., Schütte, C., and Lohse, M.J.
  (See online at https://doi.org/10.1038/s41589-020-0566-1)
• (2020) Super-resolution imaging reveals the nanoscale organization of metabotropic glutamate receptors at presynaptic active zones. Sci. Adv. 6, eaay7193
  Siddig, S., Aufmkolk, S., Doose, S., Jobin, M.L., Werner, C., Sauer, M., and Calebiro, D.
  (See online at https://doi.org/10.1126/sciadv.aay7193)
• (2020) Syndapin I Loss-of-Function in Mice Leads to Schizophrenia-Like Symptoms. Cereb. Cortex 30, 4306-4324
  Koch, N., Koch, D., Krueger, S., Tröger, J., Sabanov, V., Ahmed, T., McMillan, L.E., Wolf, D., Montag, D., Kessels, M.M., Balschun, D., and Qualmann, B.
  (See online at https://doi.org/10.1093/cercor/bhaa013)
• (2020) The Janus face of GABAergic synaptic transmission during brain development. J. Physiol. 598, 1801-1802
  Holthoff K.
  (See online at https://doi.org/10.1113/jp279623)
• (2020) Tracking down the molecular architecture of the synaptonemal complex by expansion microscopy. Nat. Commun. 11, 3222
  Zwettler, F.U., Spindler, M.C., Reinhard, S., Klein, T., Kurz, A., Benavente, R., and Sauer, M.
  (See online at https://doi.org/10.1038/s41467-020-17017-7)
• (2020) Whole-cell imaging of plasma membrane receptors by 3D lattice light-sheet dSTORM. Nat. Commun. 11, 887
  Wäldchen, F., Schlegel, J., Götz, R., Luciano, M., Schnermann, M., Doose, S., and Sauer M.
  (See online at https://doi.org/10.1038/s41467-020-14731-0)
• Antinociceptive modulation by the adhesion GPCR CIRL promotes mechanosensory signal discrimination. eLife 9, e56738
  Dannhäuser, S., Lux, T.J., Hu, C., Selcho, M., Chen, J.T., Ehmann, N., Sachidanandan, D., Stopp, S., Pauls, D., Pawlak, M., Langenhan, T., Soba, P., Rittner, H.L., and Kittel, R.J.
  (See online at https://doi.org/10.7554/eLife.56738)