Internalization of D1 dopamine receptor in striatal neurons in vivo as evidence of activation by dopamine agonists.

To investigate how dopamine influences the subcellular localization of the dopamine receptors in the striatal dopaminoceptive neurons, we have used immunohistochemistry to detect D1 dopamine receptors (D1R) after modifications of the dopamine environment. In normal rats, D1R are located mostly extrasynaptically at the plasma membrane of the cell bodies, dendrites, and spines. The intrastriatal injection of the full D1R agonist SKF-82958 and the intraperitoneal injection of the same molecule or of amphetamine (which induces a massive release of dopamine in the striatum) induce modifications of the pattern of D1R immunoreactivity in the dorsal and ventral striatum. Whereas normal rats display homogenous staining of the neuropile with staining of the plasma membrane of the cell bodies, either treatment provokes the appearance of an intense immunoreactivity in the cytoplasm and the proximal dendrites. The labeling pattern is heterogeneous and more intense in the striosomes than in the matrix. Analysis of semithin sections and electron microscopy studies demonstrates a translocation of the labeling from the plasma membrane to endocytic vesicles and endosomes bearing D1R immunoreactivity in the cytoplasm of cell bodies and dendrites. Injection of D1R antagonist (SCH-23390) alone or injection of D1R antagonist, together with amphetamine or SKF-82958, do not provoke modification of the immunoreactivity, as compared with normal rat. Our results demonstrate that, in vivo, the acute activation of dopamine receptors by direct agonists or endogenously released dopamine provokes dramatic modifications of their subcellular distribution in neurons, including internalization in the endosomal compartment in the cytoplasm. This suggests that modifications of the localization of neurotransmitter receptors, including extrasynaptic ones, may be a critical event that contributes to the postsynaptic response in vivo.