Ali Salahpour^*, Amy J. Ramsey^*, Ivan O. Medvedev^*, Brian Kile, Tatyana D. Sotnikova^*, Ericka Holmstrand, Valentina Ghisi^*, Peter J. Nicholls^*, Ling Wong, Karen Murphy^*, Susan R. Sesack, R. Mark Wightman, Raul R. Gainetdinov^*, and Marc G. Caron^*,

*Department of Cell Biology, Duke University Medical Center, Durham, NC 27710; Department of Chemistry and Neuroscience Center, University of North Carolina, Chapel Hill, NC 27599-3290; and Departments of Neuroscience and Psychiatry, University of Pittsburgh, Pittsburgh, PA 15260

Edited by Susan G. Amara, University of Pittsburgh School of Medicine, Pittsburgh, PA, and approved January 22, 2008 (received for review August 13, 2007)

Abstract

The dopamine transporter (DAT) plays a key role in the regulation of dopaminergic signaling wherein it controls both the spatial and temporal actions of dopamine. Here we evaluated the behavioral and neuRochemical consequences of increased DAT function by generating DAT transgenic mice (DAT-tg) that overexpress the transporter. These mice were generated by pronuclear injection of a bacterial artificial chromosome containing the mouse DAT locus, yielding an anatomical expression pattern of DAT-tg identical to WT. In DAT-tg mice there is a 3-fold increase in the levels of total and membrane-expressed DAT, but synaptic plasma membrane fractions of DAT-tg mice show only a 30% increase in transporter levels. Functional studies reveal that in the DAT-tg animals there is a 50% increase in the rate of dopamine (DA) uptake resulting in extracellular levels of DA that are decreased by 40%. Behaviorally, DAT-tg animals display similar locomotor stimulation when treated with DAT blockers such as GBR12909, methylphenidate, and cocaine. However, these mice demonstrate markedly increased locomotor responses to amphetamine compared with WT animals. Furthermore, compared with controls, there is a 3-fold greater increase in the amount of DA released by amphetamine in DAT-tg mice that correlates with the 3-fold increase in protein expression. Finally, DAT-tg animals show reduced operant responding for natural reward while displaying preference for amphetamine at much lower doses (0.2 and 0.5 mg/kg) than WT mice (2 mg/kg). These results suggest that overexpression of DAT leads to a marked increase in sensitivity to psychomotor and rewarding properties of amphetamine.