D amphetamine hemisulfate salt

Animals were trained on the risk-discounting task for 15 days before the effects of d-amphetamine and flupenthixol on risk discounting were assessed. Different doses of d-amphetamine hemisulfate salt (Sigma Aldrich) and cis-(Z)-Flupenthixol dihydrochloride (Sigma Aldrich) were investigated in two subsequent experiments. In the first experiment, higher drug doses (amphetamine 2mg/kg, flupenthixol 0.4mg/kg vs. saline 1ml/kg) were tested; in the second experiment, lower drug doses (amphetamine 0.5mg/kg, flupenthixol 0.25mg/kg vs. saline 1ml/kg) were tested. Drugs were dissolved in physiological saline and administered intraperitoneally at a volume of 1ml/kg. After administration rats were returned to their home cages for 30min (amphetamine), 45min (saline), or 60min (flupenthixol) until behavioral testing. A within-subject design was used in each experiment, with the order of drug administration pseudo-randomized across rats. Drugs (saline, amphetamine, flupenthixol) in each experiment were tested in two subsequent 4-day blocks (see Supplementary Material).

(5R,10S)-(+)-5-Methyl-10,11-dihydro-5H-dibenzo[a,d]cyclohepten-5,10-imine hydrogen maleate ((+)-MK801, 0.3 mg/kg) and haloperidol (0.5 mg/kg) were purchased from Tocris Bioscience, and D-amphetamine hemisulfate salt (2.5 mg/kg), clozapine N-oxide (CNO, 1 mg/kg), and tamoxifen (100 mg/kg) from Sigma-Aldrich. All drugs were administered intraperitoneally in a volume of 10 ml/kg and dissolved in 0.9% (w/v) NaCl (saline), except CNO that was dissolved in 0.1% DMSO and tamoxifen that was dissolved in sunflower oil/ethanol (10:1) to a final concentration of 10 mg/ml.

Cocaine hydrochloride, D-amphetamine hemisulfate salt and ZnCl₂ were from Sigma-Aldrich (St. Louis, MO). JHC1-64 was synthesized in the Medicinal Chemistry Section, NIDA-IRP as previously described^{30 (link)}. Restriction enzymes are from New England Biolabs (Ipswich, MA). All other chemicals were from Thermo Fisher Scientific (Pittsburgh, PA) or Sigma-Aldrich.

Monensin sodium salt and D-amphetamine hemisulfate salt were purchased from Sigma-Aldrich Co.
The substrate/efflux experiments were performed as described before by [29 (link)]. Briefly, HEK-DAT cells were grown in 5 mm diameter PDL-coated coverslips. Cells were incubated with 0.05 μM [³H]MPP⁺ at 37°C for 20 min. The coverslips were transferred onto superfusion chambers (0.2 ml) and excess radioactivity was washed out with KHB for 40 min (0.7 ml/min) at 25°C to obtain stable baselines. The experiment was started with the collection of fractions (2 min) as depicted in Fig 2. During the experiments the buffer was switched either to Monensin or remained at control buffer after the collection of three baseline fractions for another four fractions. Subsequently, R-MO or D-amphetamine was added for another five fractions as indicated in the figure. Finally, the remaining radioactivity was collected by treatment with 1% SDS.