Molecular cloning of the mouse dopamine transporter and pharmacological comparison with the human homologue

Drug abuse is a serious problem in the United States and in the world. Cocaine and amphetamines, widely used drugs of abuse, bind to dopamine (DA), serotonin, and norepinephrine transporters with high affinity and block their functions. It is believed that the dopamine transporter plays a key role in the mechanism of cocaine addiction. Because a good portion of our knowledge about drug addiction is derived from studying mouse as an animal model, it is essential to compare the properties of dopamine transporter from human and mouse. We report here the cloning of the mouse dopamine transporter (mDAT) cDNA and its expression and comparison with the human DAT. The 3.4 kilobase (kb) cDNA encodes a polypeptide that is 93.5% identical to the hDAT, with 619 amino acid residues and a calculated molecular weight of 68.8kDa. Dopamine transporters from mouse and human were stably expressed in the same parental MDCK cells and their properties were compared. The Michaelis-Menten constant Km values are 2.0 microM for mDAT and 2.4 microM for hDAT. Mouse and human DAT were also compared for drug inhibition profiles. Dopamine transporters from the two species have the same sensitivity to amphetamine (Kd: 0.75 microM) and bupropion (Kd: 1.5 microM). However, hDAT is more sensitive than mDAT to cocaine (Kd: 0.14 microM and 0. 29 microM respectively) and to ritalin (Kd: 0.038 microM and 0. 12 microM respectively). The cloning of mDAT cDNA provides an important tool for further study of the mechanism of drug addiction using mouse as an animal model.