Trafficking of the Dopamine Transporter

Berelashvili, Lali

Student Work

Trafficking of the Dopamine Transporter

Público

Dysregulation of dopamine is indicated in multiple neurological disorders, including ADHD, addiction, and Parkinson’s. The dopamine transporter (DAT) is one of the most important components of regulating extracellular dopamine levels. Typically, DAT is trafficked at a steady state. Previous research has shown that insertion of DAT into the cell membrane is stimulated by D2 activation. However, mutants such as DAT R615C are less stable on the surface and may facilitate dysregulated reuptake of dopamine into the presynaptic neuron. This may suggest that arginine at location DAT 615 is necessary for insertion of DAT into the cell membrane. To determine whether arginine is required for steady DAT trafficking, the mutant mDAT R615C was engineered and cultivated in human embryonic kidney cells. Immunoblotting was used to determine expression of cells with mDAT R615C, compared to wild-type mDAT. Trafficking was analyzed through biotinylation and immunoblotting. It was found that DAT R615C expresses 44% as much as wild-type DAT. Further, a significant 44% decrease in surface DAT was observed in DAT R615C compared to the wild-type. This suggests that DAT R615C is an unstable mutant of the dopamine transporter and may affect dopamine dependent behaviors. Findings support that arginine at DAT 615 is required for steady trafficking.

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