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Development of high-specificity fluorescent probes to enable cannabinoid type 2 receptor studies in living cells

Item Type:Article
Title:Development of high-specificity fluorescent probes to enable cannabinoid type 2 receptor studies in living cells
Creators Name:Sarott, R.C. and Westphal, M. and Pfaff, P. and Korn, C. and Sykes, D.A. and Gazzi, T. and Brennecke, B. and Atz, K. and Weise, M. and Mostinski, Y. and Hompluem, P. and Koers, E. and Miljus, T. and Roth, N.J. and Asmelash, H. and Vong, M.C. and Piovesan, J. and Guba, W. and Rufer, A. and Kusznir, E.A. and Huber, S. and Raposo, C. and Zirwes, E.A. and Osterwald, A. and Pavlovic, A. and Moes, S. and Beck, J. and Benito-Cuesta, I. and Grande, T. and Ruiz de Martin, S. and Yeliseev, A.A. and Drawnel, F. and Widmer, G. and Holzer, D. and van der Wel, T. and Mandhair, H. and Yuan, C.Y. and Drobyski, W. and Saroz, Y. and Grimsey, N.L. and Honer, M. and Fingerle, J. and Gawrisch, K. and Romero, J. and Hillard, C. and Varga, Z. and van der Stelt, M. and Pacher, P. and Gertsch, J. and McCormick, P. and Ullmer, C. and Oddi, S. and Maccarrone, M. and Veprintsev, D. and Nazaré, M. and Grether, U. and Carreira, E.M.
Abstract:Pharmacological modulation of cannabinoid type 2 receptor (CB2R) holds promise for the treatment of numerous con-ditions, including inflammatory diseases, autoimmune disorders, pain, and cancer. Despite the significance of this re-ceptor, researchers lack reliable tools to address questions concerning the expression and complex mechanism of CB2R signaling, especially in cell-type and tissue-dependent context. Herein, we report for the first time a versatile ligand platform for the modular design of a collection of highly specific CB2R fluorescent probes, used successfully across ap-plications, species and cell types. These include flow cytometry of endogenously expressing cells, real-time confocal microscopy of mouse splenocytes and human macrophages, as well as FRET-based kinetic and equilibrium binding assays. High CB2R specificity was demonstrated by competition experiments in living cells expressing CB2R at native levels. The probes were effectively applied to FACS analysis of microglial cells derived from a mouse model relevant to Alzheimer's disease.
Keywords:Alzheimer Disease, Animal Disease Models, CB2 Cannabinoid Receptor, CHO Cells, Flow Cytometry, Fluorescence Resonance Energy Transfer, Fluorescent Dyes, Ligands, Microglia, Molecular Docking Simulation, Molecular Probes, Optical Imaging, Sensitivity and Specificity, Signal Transduction, Animals, Cricetulus, Mice
Source:Journal of the American Chemical Society
Publisher:American Chemical Society
Page Range:16953-16964
Date:7 October 2020
Official Publication:https://doi.org/10.1021/jacs.0c05587
PubMed:View item in PubMed

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