Molecular cloning and functional expression of the mouse dopamine transporter.

The full coding region of the murine dopamine transporter (mDAT) cDNA was cloned by PCR with a sense primer derived from the partial mDAT gene sequence and an antisense primer deduced from the rat dopamine transporter cDNA. The mDAT cDNA encodes a typical member of the family of Na+- and Cl--dependent neurotransmitter transporters with 99.2; 93.4 and 85.4% amino acid identity to the rat, human and bovine DATs, respectively. Functional expression of the mDAT cDNA in transiently transfected human embryonic kidney (HEK293) cells exhibited the typical pharmacological features of a dopamine transporter. [3H]dopamine uptake through the mDAT was inhibited with high potency by GBR12909 (IC50 = 5.2 nM) and not significantly affected by 100 nM desipramine. [3H]dopamine uptake also was inhibited through increasing concentrations of dopamine (IC50 = 0.93 microM) or 1-methyl-4-phenylpyridinium (MPP+; IC50 = 13.2microM).