In vitro and in vivo characterization of the agonist-dependent D3 dopamine receptor tolerance property

The D3 dopamine receptor has the highest affinity for dopamine, many antipsychotics as well as agonists used to treat Parkinson's disease and related disorders. We and others have reported that the D3 receptor exhibits a tolerance property wherein repeated agonist stimulation of the receptor results in a progressive loss of agonist-induced signaling response. Recently we reported that the D3 receptor tolerance property is agonist dependent and identified a novel agonist, ES609, which does not elicit D3 receptor tolerance. Here, we used the classical tolerance-inducing D3 receptor agonist, PD128907, and the novel agonist, ES609, to demonstrate that the D3 receptor tolerance property is exhibited not only in cellular signaling in vitro and in vivo, but also manifests at the behavior level. Using AtT-20 cells stably expressing D3 receptors we show that PD128907, but not ES609, induces tolerance in the D3 receptor-mitogen activated protein kinase (MAPK) pathway. Using the novel drd3-EGFP reporter mice, we demonstrate that 0.05 mg/kg PD128907 and 10 mg/kg ES609 selectively activate the D3 receptor-MAPK signaling pathway in vivo; however, only PD128907 induces tolerance. Locomotor behavior assessment showed that both PD128907 and ES609 decreased locomotor activity of the drd3-EGFP mice. While the agonist-induced decrease in locomotor activity was attenuated in drd3-EGFP mice administered two sequential doses of tolerance-inducing agonist PD128907, this attenuation was not seen in mice repeatedly administered the novel agonist, ES609. Together the results suggest that the D3 receptor tolerance property is exhibited in MAPK signaling in vitro and in vivo and also affects agonist-induced locomotor behavior.