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

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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., Westphal, M., Pfaff, P., Korn, C., Sykes, D.A., Gazzi, T., Brennecke, B., Atz, K., Weise, M., Mostinski, Y., Hompluem, P., Koers, E., Miljus, T., Roth, N.J., Asmelash, H., Vong, M.C., Piovesan, J., Guba, W., Rufer, A., Kusznir, E.A., Huber, S., Raposo, C., Zirwes, E.A., Osterwald, A., Pavlovic, A., Moes, S., Beck, J., Benito-Cuesta, I., Grande, T., Ruiz de Martin, S., Yeliseev, A.A., Drawnel, F., Widmer, G., Holzer, D., van der Wel, T., Mandhair, H., Yuan, C.Y., Drobyski, W., Saroz, Y., Grimsey, N.L., Honer, M., Fingerle, J., Gawrisch, K., Romero, J., Hillard, C., Varga, Z., van der Stelt, M., Pacher, P., Gertsch, J., McCormick, P., Ullmer, C., Oddi, S., Maccarrone, M., Veprintsev, D., Nazaré, M., 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
ISSN:	0002-7863
Publisher:	American Chemical Society
Volume:	142
Number:	40
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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