Prolonged treatment with pramipexole promotes physical interaction of striatal dopamine D3 autoreceptors with dopamine transporters to reduce dopamine uptake

• Prolonged PPX treatment induces physical interaction between D3 autoreceptor and DAT.
• PPX also promotes DAT dimer formation and DAT interaction with D2R and α-syn.
• Changes in DAT interactions are associated with a decrease in DAT affinity for DA.
• The changes induced by PPX in DAT interactions and function are D3R-dependent.

The dopamine (DA) transporter (DAT), a membrane glycoprotein expressed in dopaminergic neurons, clears DA from extracellular space and is regulated by diverse presynaptic proteins like protein kinases, α-synuclein, D2 and D3 autoreceptors. DAT dysfunction is implicated in Parkinson's disease and depression, which are therapeutically treated by dopaminergic D2/D3 receptor (D2/D3R) agonists. It is, then, important to improve our understanding of interactions between D3R and DAT. We show that prolonged administration of pramipexole (0.1 mg/kg/day, 6 to 21 days), a preferential D3R agonist, leads to a decrease in DA uptake in mouse striatum that reflects a reduction in DAT affinity for DA in the absence of any change in DAT density or subcellular distribution. The effect of pramipexole was absent in mice with genetically-deleted D3R (D3R−/−), yet unaffected in mice genetically deprived of D2R (D2R−/−). Pramipexole treatment induced a physical interaction between D3R and DAT, as assessed by co-immunoprecipitation and in situ proximity ligation assay. Furthermore, it promoted the formation of DAT dimers and DAT association with both D2R and α-synuclein, effects that were abolished in D3R−/− mice, yet unaffected in D2R−/− mice, indicating dependence upon D3R. Collectively, these data suggest that prolonged treatment with dopaminergic D3 agonists provokes a reduction in DA reuptake by dopaminergic neurons related to a hitherto-unsuspected modification of the DAT interactome. These observations provide novel insights into the long-term antiparkinson, antidepressant and additional clinical actions of pramipexole and other D3R agonists.

Schematic drawing summarizing the effects of prolonged PPX treatment (0.1 mg/kg/day; 6 days) on DAT at dopaminergic terminals in mouse striatum. PPX promotes the formation of DAT dimers, and the physical interaction of DAT with its protein partners α-synuclein, D2 and D3 autoreceptors. These changes are associated with a decline in dopamine uptake, resulting in an increase in extracellular dopamine and a decrease in intracellular dopamine. Extra, extracellular space; Intra, intracellular space.Figure optionsDownload as PowerPoint slide