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Expression of a Human Cocaine-Sensitive Dopamine Transporter in Xenopus laevis Oocytes
Authors:Michael J. Bannon  Chang-Hong Xue  Kazuhiko Shibata  Ljubisa J. Dragovic  Gregory Kapatos
Affiliation:Center for Cell Biology, Sinai Research Institute, Detroit, Michigan.
Abstract:The human dopamine transporter was expressed in Xenopus laevis oocytes following injection of mRNA isolated from human brain substantia nigra. The specific accumulation of [3H]dopamine into these oocytes was time and Na+ dependent. Furthermore, [3H]dopamine accumulation was prevented by coincubation of oocytes with dopamine (100 microM) or with the dopamine uptake inhibitors GBR 12909 (1 microM) or cocaine (3 microM). In contrast, oocyte injection of mRNA isolated from human globus pallidus, an area devoid of dopamine neuron perikarya, did not elicit expression of the dopamine transporter. Oocyte expression of the human dopamine transporter can be used for the further characterization and cloning of this low-abundance membrane protein.
Keywords:Dopamine transporter    Oocyte    Uptake    Cocaine    mRNA
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