Detection of cannabinoid receptor type 2 in native cells and zebrafish with a highly potent, cell-permeable fluorescent probe. Issue 19 (22nd April 2022)

• Record Type: Journal Article
• Authors: Gazzi, Thais; Brennecke, Benjamin; Atz, Kenneth; Korn, Claudia; Sykes, David; Forn-Cuni, Gabriel; Pfaff, Patrick; Sarott, Roman C.; Westphal, Matthias V.; Mostinski, Yelena; Mach, Leonard; Wasinska-Kalwa, Malgorzata; Weise, Marie; Hoare, Bradley L.; Miljuš, Tamara; Mexi, Maira; Roth, Nicolas; Koers, Eline J.; Guba, Wolfgang; Alker, André; Rufer, Arne C.; Kusznir, Eric A.; Huber, Sylwia; Raposo, Catarina; Zirwes, Elisabeth A.; Osterwald, Anja; Pavlovic, Anto; Moes, Svenja; Beck, Jennifer; Nettekoven, Matthias; Benito-Cuesta, Irene; Grande, Teresa; Drawnel, Faye; Widmer, Gabriella; Holzer, Daniela; van der Wel, Tom; Mandhair, Harpreet; Honer, Michael; Fingerle, Jürgen; Scheffel, Jörg; Broichhagen, Johannes; Gawrisch, Klaus; Romero, Julián; Hillard, Cecilia J.; Varga, Zoltan V.; van der Stelt, Mario; Pacher, Pal; Gertsch, Jürg; Ullmer, Christoph; McCormick, Peter J.; Oddi, Sergio; Spaink, Herman P.; Maccarrone, Mauro; Veprintsev, Dmitry B.; Carreira, Erick M.; Grether, Uwe; Nazaré, Marc;
• Is Part Of: Chemical science. Volume 13:Issue 19(2022)
• Page Start: 5539
• Page End: 5545
• Publication Date: 2022-04-22
• Subjects: Chemistry -- Periodicals; 540.5
• DOI: 10.1039/d1sc06659e
• Languages: English
• ISSNs: 2041-6520

Abstract

Abstract : Detection and visualization of the cannabinoid receptor type 2 by a cell-permeable high affinity fluorescent probe platform enables tracing receptor trafficking in live cells and in zebrafish. Abstract : Despite its essential role in the (patho)physiology of several diseases, CB2 R tissue expression profiles and signaling mechanisms are not yet fully understood. We report the development of a highly potent, fluorescent CB2 R agonist probe employing structure-based reverse design. It commences with a highly potent, preclinically validated ligand, which is conjugated to a silicon-rhodamine fluorophore, enabling cell permeability. The probe is the first to preserve interspecies affinity and selectivity for both mouse and human CB2 R. Extensive cross-validation (FACS, TR-FRET and confocal microscopy) set the stage for CB2 R detection in endogenously expressing living cells along with zebrafish larvae. Together, these findings will benefit clinical translatability of CB2 R based drugs.