Background and purpose
The CB(1) cannabinoid receptor and the beta(2)-adrenoceptor are G protein-coupled receptors (GPCRs) co-expressed in many tissues. The present study examined physical and functional interactions between these receptors in a heterologous expression system and in primary human ocular cells. Experimental approach: Physical interactions between CB(1) receptors and beta(2)-adrenoceptors were assessed using bioluminescence resonance energy transfer (BRET). Functional interactions between these receptors were evaluated by examining receptor trafficking, as well as extracellular signal-regulated kinase (ERK) and cyclic AMP response element binding protein (CREB) signalling. Key
Purpose
The CB(1) cannabinoid receptor and the beta(2)-adrenoceptor are G protein-coupled receptors (GPCRs) co-expressed in many tissues. The present study examined physical and functional interactions between these receptors in a heterologous expression system and in primary human ocular cells. Experimental approach: Physical interactions between CB(1) receptors and beta(2)-adrenoceptors were assessed using bioluminescence resonance energy transfer (BRET). Functional interactions between these receptors were evaluated by examining receptor trafficking, as well as extracellular signal-regulated kinase (ERK) and cyclic AMP response element binding protein (CREB) signalling. Key
Results
Physical interactions between CB(1) receptors and beta(2)-adrenoceptors were demonstrated using BRET. In human embryonic kidney (HEK) 293H cells, co-expression of beta(2)-adrenoceptors tempered the constitutive activity and increased cell surface expression of CB(1) receptors. Co-expression altered the signalling properties of CB(1 )receptors, resulting in increased Galpha(i)-dependent ERK phosphorylation, but decreased non-Galpha(i)-mediated CREB phosphorylation. The CB(1) receptor inverse agonist AM251 (N-(piperidin-1-yl)-5-(4-iodophenyl)-1-(2,4-dichlorophenyl)-4-methyl-1H-pyrazole-3-carboxamide) attenuated beta(2)-adrenoceptor-pERK signalling in cells expressing both receptors, while the CB(1) receptor neutral antagonist O-2050 ((6aR,10aR)-3-(1-methanesulfonylamino-4-hexyn-6-yl)-6a,7,10,10a-tetrahydro-6,6,9-trimethyl-6H-dibenzo[b,d]pyran) did not. The actions of AM251 and O-2050 were further examined in primary human trabecular meshwork (HTM) cells, which are ocular cells endogenously co-expressing CB(1) receptors and beta(2)-adrenoceptors. In HTM cells, as in HEK 293H cells, AM251 but not O-2050, altered the beta(2)-adrenoceptor-pERK response.