To investigate the influence of neurotransmitter on G-protein-coupled receptor trafficking and compartimentalization in neurons, we have developed a model of primary neuronal cultures from fetal rat striatum on which we have studied the cellular and subcellular distribution and trafficking of the D₁ dopaminergic receptor. This receptor is known to be somatodendritic and axonal targeted in vivo, mostly to extrasynaptic locations. Immunohistochemical studies at the light and electron microscopic levels showed that, in cultures, the D₁ dopaminergic receptor is expressed in the absence of dopamine stimulation. The pattern of D₁ dopaminergic receptor immunostaining after stimulation by the D₁ dopaminergic receptor agonist SKF 82958 (1μM) is dramatically modified with a decrease of the number of labeled D₁ dopaminergic receptor puncta (−40%) and an increase of their size in both dendrites (+120%) and axons (+240%). Seven hours after removal of the agonist, return to normal pattern was observed. The D₁ dopaminergic receptor antagonist SCH 23390 (2μM) abolishes the effect of SKF 82958. Electron microscopy demonstrated, in dendrites, a translocation of the labeling from the plasma membrane to endosomes. Axonal D₁ dopaminergic receptor redistribution after acute stimulation indicates that the D₁ dopaminergic receptor is membrane targeted and responsive to stimulation. These results validate primary culture of striatal neurons to study subcellular localization and intraneuronal trafficking of G-protein-coupled receptors. This preparation will be useful to address various questions concerning the behavior and the trafficking of these receptors in neurons in relation to the neurotransmitter environment.