Identifying nonaddictive opioid medications is a high priority in medical science, but μ-opioid receptors (MORs) mediate both the analgesic and addictive effects of opioids. We found a significant pharmacodynamic difference between morphine and methadone that is determined entirely by heteromerization of MORs with galanin Gal1 receptors (Gal1Rs), rendering a profound decrease in the potency of methadone. This finding was explained by the weaker proficiency of methadone in activating the dopaminergic system as compared with morphine and predicted a dissociation of the therapeutic and euphoric effects of methadone, which was corroborated by a significantly lower incidence of self-reports of feeling “high” in methadone-medicated patients. These results suggest that μ-opioid–Gal1R heteromers mediate the dopaminergic effects of opioids. The results further suggest a lower addictive liability of some opioids, such as methadone, due to their selective low potency for the μ-opioid–Gal1R heteromer.

The poor norepinephrine innervation and high density of Gi/o-coupled α_2A- and α_2C-adrenoceptors in the striatum and the dense striatal dopamine innervation have prompted the possibility that dopamine could be an effective adrenoceptor ligand. Nevertheless, the reported adrenoceptor agonistic properties of dopamine are still inconclusive. In this study, we analyzed the binding of norepinephrine, dopamine, and several compounds reported as selective dopamine D₂-like receptor ligands, such as the D₃ receptor agonist 7-OH-PIPAT and the D₄ receptor agonist RO-105824, to α₂-adrenoceptors in cortical and striatal tissue, which express α_2A-adrenoceptors and both α_2A- and α_2C-adrenoceptors, respectively. The affinity of dopamine for α₂-adrenoceptors was found to be similar to that for D₁-like and D₂-like receptors. Moreover, the exogenous dopamine receptor ligands also showed high affinity for α_2A- and α_2C-adrenoceptors. Their ability to activate Gi/o proteins through α_2A- and α_2C-adrenoceptors was also analyzed in transfected cells with bioluminescent resonance energy transfer techniques. The relative ligand potencies and efficacies were dependent on the Gi/o protein subtype. Furthermore, dopamine binding to α₂-adrenoceptors was functional, inducing changes in dynamic mass redistribution, adenylyl cyclase activity, and ERK1/2 phosphorylation. Binding events were further studied with computer modeling of ligand docking. Docking of dopamine at α_2A- and α_2C-adrenoceptors was nearly identical to its binding to the crystallized D₃ receptor. Therefore, we provide conclusive evidence that α_2A- and α_2C-adrenoceptors are functional receptors for norepinephrine, dopamine, and other previously assumed selective D₂-like receptor ligands, which calls for revisiting previous studies with those ligands.

Polymorphic alleles of the human dopamine D₄ receptor gene (DRD4) have been consistently associated with individual differences in personality traits and neuropsychiatric disorders, particularly between the gene encoding dopamine D_4.7 receptor variant and attention deficit hyperactivity disorder (ADHD). The α_2A adrenoceptor gene has also been associated with ADHD. In fact, drugs targeting the α_2A adrenoceptor (α_2AR), such as guanfacine, are commonly used in ADHD treatment. In view of the involvement of dopamine D₄ receptor (D₄R) and α_2AR in ADHD and impulsivity, their concurrent localization in cortical pyramidal neurons and the demonstrated ability of D₄R to form functional heteromers with other G protein-coupled receptors, in this study we evaluate whether the α_2AR forms functional heteromers with D₄R and weather these heteromers show different properties depending on the D₄R variant involved. Using cortical brain slices from hD_4.7R knock-in and wild-type mice, here, we demonstrate that α_2AR and D₄R heteromerize and constitute a significant functional population of cortical α_2AR and D₄R. Moreover, in cortical slices from wild-type mice and in cells transfected with α_2AR and D_4.4R, we detect a negative crosstalk within the heteromer. This negative crosstalk is lost in cortex from hD_4.7R knock-in mice and in cells expressing the D_4.7R polymorphic variant. We also show a lack of efficacy of D₄R ligands to promote G protein activation and signaling only within the α_2AR-D_4.7R heteromer. Taken together, our results suggest that α_2AR-D₄R heteromers play a pivotal role in catecholaminergic signaling in the brain cortex and are likely targets for ADHD pharmacotherapy.

The functional role of the dopamine D₄ receptor (D₄R) and its main polymorphic variants has become more evident with the demonstration of heteromers of D₄R that control the function of frontal cortico-striatal neurons. Those include heteromers with the α_2A adrenoceptor (α_2AR) and with the D₂R, localized in their cortical somato-dendritic region and striatal nerve terminals, respectively. By using biophysical and cell-signaling methods and heteromer-disrupting peptides in mammalian transfected cells and rat brain slice preparations, here we provide evidence for a new functionally relevant D₄R heteromer, the α_1AR-D₄R heteromer, which is also preferentially localized in cortico-striatal glutamatergic terminals. Significant differences in allosteric modulations between heteromers of α_1AR with the D_4.4R and D_4.7R polymorphic variants could be evidenced with the analysis of G protein-dependent and independent signaling. Similar negative allosteric modulations between α_1AR and D₄R ligands could be demonstrated for both α_1AR-D_4.4R and α_1AR-D_4.7R heteromers on G protein-independent signaling, but only for α_1AR-D_4.4R on G protein-dependent signaling. From these functional differences, it is proposed that the D_4.4R variant provides a gain of function of the α_1AR-mediated noradrenergic stimulatory control of cortico-striatal glutamatergic neurotransmission, which could result in a decrease in the vulnerability for impulse control-related neuropsychiatric disorders and increase in the vulnerability for posttraumatic stress disorder.