Detection of cannabinoid receptor type 2 in native cells and zebrafish with a highly potent, cell-permeable fluorescent probe.
Thais GazziBenjamin BrenneckeKenneth AtzClaudia KornDavid SykesGabriel Forn-CuniPatrick PfaffRoman C SarottMatthias V WestphalYelena MostinskiLeonard MachMalgorzata Wasinska-KalwaMarie WeiseBradley L HoareTamara MiljušMaira MexiNicolas RothEline J KoersWolfgang GubaAndré AlkerArne C RuferEric A KusznirSylwia HuberCatarina RaposoElisabeth A ZirwesAnja OsterwaldAnto PavlovicSvenja MoesJennifer BeckMatthias NettekovenIrene Benito-CuestaTeresa GrandeFaye DrawnelGabriella WidmerDaniela HolzerTom van der WelHarpreet K MandhairMichael HonerJürgen FingerleJörg ScheffelJohannes BroichhagenKlaus GawrischJulián RomeroCecilia J HillardZoltan V VargaMario van der SteltPal PacherJürg GertschChristoph UllmerPeter J McCormickSergio OddiHerman P SpainkMauro MaccarroneDmitry B VeprintsevErick M CarreiraUwe GretherMarc NazarePublished in: Chemical science (2022)
Despite its essential role in the (patho)physiology of several diseases, CB 2 R tissue expression profiles and signaling mechanisms are not yet fully understood. We report the development of a highly potent, fluorescent CB 2 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 CB 2 R. Extensive cross-validation (FACS, TR-FRET and confocal microscopy) set the stage for CB 2 R detection in endogenously expressing living cells along with zebrafish larvae. Together, these findings will benefit clinical translatability of CB 2 R based drugs.