Molecular cloning and functional expression of the mouse dopamine transporter - Rapid communication

The full coding region of the murine dopamine transporter (mDAT) cDNA was c loned by PCR with a sense primer derived from the partial mDAT gene sequenc e and an antisense primer deduced from the rat dopamine transporter cDNA. T he mDAT cDNA encodes a typical member of the family of Na+- and Cl-- depend ent neurotransmitter transporters with 99.2; 93.4 and 85.4% amino acid iden tity 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 transp orter. [H-3]dopamine uptake through the mDAT was inhibited with high potenc y by GBR12909 (IC50 = 5.2 nM) and not significantly affected by 100nM desip ramine. [H-3]dopamine uptake also was inhibited through increasing concentr ations of dopamine (IC50 = 0.93 mu M) or 1-methyl-4-phenylpyridinium (MPP+; IC50 = 13.2 mu M).