Faculty Collaboration Database

Development of High-Specificity Fluorescent Probes to Enable Cannabinoid Type 2 Receptor Studies in Living Cells. J Am Chem Soc 2020 Oct 07;142(40):16953-16964

Date

09/10/2020

Pubmed ID

32902974

DOI

10.1021/jacs.0c05587

Scopus ID

2-s2.0-85092682450 (requires institutional sign-in at Scopus site)   31 Citations

Abstract

Pharmacological modulation of cannabinoid type 2 receptor (CB₂R) holds promise for the treatment of numerous conditions, including inflammatory diseases, autoimmune disorders, pain, and cancer. Despite the significance of this receptor, researchers lack reliable tools to address questions concerning the expression and complex mechanism of CB₂R signaling, especially in cell-type and tissue-dependent contexts. 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 applications, 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.

Author List

Sarott RC, Westphal MV, Pfaff P, Korn C, Sykes DA, Gazzi T, Brennecke B, Atz K, Weise M, Mostinski Y, Hompluem P, Koers E, Miljuš T, Roth NJ, Asmelash H, Vong MC, Piovesan J, Guba W, Rufer AC, Kusznir EA, Huber S, Raposo C, Zirwes EA, Osterwald A, Pavlovic A, Moes S, Beck J, Benito-Cuesta I, Grande T, Ruiz de Martı N Esteban S, Yeliseev A, Drawnel F, Widmer G, Holzer D, van der Wel T, Mandhair H, Yuan CY, Drobyski WR, Saroz Y, Grimsey N, Honer M, Fingerle J, Gawrisch K, Romero J, Hillard CJ, Varga ZV, van der Stelt M, Pacher P, Gertsch J, McCormick PJ, Ullmer C, Oddi S, Maccarrone M, Veprintsev DB, Nazaré M, Grether U, Carreira EM

Authors

William R. Drobyski MD Professor in the Medicine department at Medical College of Wisconsin
Cecilia J. Hillard PhD Associate Dean, Center Director, Professor in the Pharmacology and Toxicology department at Medical College of Wisconsin

MESH terms used to index this publication - Major topics in bold

Alzheimer Disease
Animals
CHO Cells
Cricetulus
Disease Models, Animal
Flow Cytometry
Fluorescence Resonance Energy Transfer
Fluorescent Dyes
Humans
Ligands
Mice
Microglia
Molecular Docking Simulation
Molecular Probes
Optical Imaging
Receptor, Cannabinoid, CB2
Sensitivity and Specificity
Signal Transduction