Antipsychotic-Like Efficacy of Dopamine D₂ Receptor-Biased Ligands is Dependent on Adenosine A_2A Receptor Expression

Dopamine D₂ receptor (D₂R) activation triggers both G protein- and β-arrestin-dependent signaling. Biased D₂R ligands activating β-arrestin pathway have been proposed as potential antipsychotics. The ability of D₂R to heteromerize with adenosine A_2A receptor (A_2AR) has been associated to D₂R agonist-induced β-arrestin recruitment. Accordingly, here we aimed to demonstrate the A_2AR dependence of D₂R/β-arrestin signaling. By combining bioluminescence resonance energy transfer (BRET) between β-arrestin-2 tagged with yellow fluorescent protein and bimolecular luminescence complementation (BiLC) of D₂R/A_2AR homomers and heteromers, we demonstrated that the D₂R agonists quinpirole and UNC9994 could promote β-arrestin-2 recruitment only when A_2AR/D₂R heteromers were expressed. Subsequently, the role of A_2AR in the antipsychotic-like activity of UNC9994 was assessed in wild-type and A_2AR^-/- mice administered with phencyclidine (PCP) or amphetamine (AMPH). Interestingly, while UNC9994 reduced hyperlocomotion in wild-type animals treated either with PCP or AMPH, in A_2AR^-/- mice, it failed to reduce PCP-induced hyperlocomotion or produced only a moderate reduction of AMPH-mediated hyperlocomotion. Overall, the results presented here reinforce the notion that D₂R/A_2AR heteromerization facilitates D₂R β-arrestin recruitment, and furthermore, reveal a pivotal role for A_2AR in the antipsychotic-like activity of the β-arrestin-biased D₂R ligand, UNC9994.