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All experiments were conducted in accordance with NIH guidelines for the care and use of animals and with an approved animal protocol from the Duke University Animal Care and Use Committee. Wildtype and homozygous DAT-KO mice were derived from crossing (over seven to eight generations) heterozygous DAT C57BL6/129SvJ animals. Mice were housed four or five to a cage, maintained under standard lab conditions (12-hour light/dark cycle) with food and water provided ad libitum, and tested at 12 to 15 weeks of age. Locomotion was evaluated in an automated Omnitech Digiscan apparatus (AccuScan Instruments, Columbus, OH) under illuminated conditions. Animals were observed individually at 5-min intervals for a maximum of 4 hours. Locomotor activity was measured in terms of the total distance covered (horizontal activity), rearings were expressed in terms of the number of vertical beam breaks (vertical activity), and the stereotypy time refers to the total time that stereotypic behaviors (repetitive beam breaks of a given beam or beams with intervals less than 1 s) were observed. In the "home" cage condition, mice were housed individually in cages for 24 hours, and then the cage and mouse were placed in the Omnitech Digiscan apparatus and activity was monitored over 1 hour. The data are expressed as the total distance covered in 1 hour for the novel and home cage environments. For drug studies, mice were initially placed into the open field for 30 min, then injected with saline (10 ml/kg, ip) or the drug, and returned to the open-field apparatus. All data are presented as means and SEMs. Statistical significance of all data presented in this paper was analyzed by two-way analysis of variance followed by Duncan's tests. A P < 0.05 was considered significant.
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Microdialysate samples were collected from the right striatum 24 hours after surgery, separated, and quantitated by high-performance liquid chromatography as described earlier for freely moving mice (2, 5). In accordance with previous reports (2, 5), basal extracellular DA concentrations in the DAT-KO mice were higher than those in the wild-type controls: Concentrations of DA in diatysates were 57.3 ± 13.4 fmol/ 20 μl in the wild-type mice (n = 12) and 354.5 ± 45.9 fmol/20 μl in the DAT-KO mice (n = 15). The data are presented as means and SEMs of the percentage of change from baseline (100%) from a mean of three samples from each mouse before exposure to the novel environment or before drug or saline administration.
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This work was supported in part by grant MH-40159 from NIH and unrestricted gifts from Bristol Myers Squibb and Zeneca Pharmaceutical (to M.G.C.). M.G.C. is an Investigator of the Howard Hughes Medical Institute. R.R.G. was a recipient of a fellowship from the Tourette Syndrome Association and is a visiting researcher from the Institute of Pharmacology, Russian Academy of Medical Sciences, Baltiyskaya 8, 125315 Moscow, Russia.
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