A Receptor Mechanism for Methamphetamine Action in Dopamine Transporter Regulation in Brain

Data Supplement

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• Data Supplement - Fig. S1. Representative data showing the specific 3Hdopamine uptake (dpm) in transfected cells and in rhesus monkey and mouse striatal synaptosomes tested. Cells and synaptosomes were loaded with 10 nM 3Hdopamine alone (3HDA) or 10 nM 3Hdopamine plus 100 nM methamphetamine (3HDA+METH) for a time course of 1, 2, 3, 4, 5, 10, 20, 30 min. The counting represents the 3Hdopamine uptake in 50 ml suspension of TAAR1-DAT cells (A), DAT cells (B), wild type mouse (WT) striatal synaptosomes (C), TAAR1 knockout (TAAR1-/-) mouse striatal synaptosomes (D), and rhesus monkey striatal synaptosomes (E).
• Data Supplement - Fig. S2. An analysis of the uptake inhibition observed at the 10 min time point for the data presented in Fig. 1B and 1C reveals significantly greater inhibition of 3Hdopamine uptake at the 10 min time point in TAAR1-DAT versus DAT cells (A), and TAAR1 knockout (TAAR1-/-) mouse synaptosomes versus rhesus monkey (Rhesus) or wild type (WT) mouse synaptosomes (B). The reduction in 3Hdopamine uptake at the 10 min time point (difference in the percentage uptake values between 3HDA and 3HDA+METH) is graphed in each case. Data shown are values of mean � SEM for three independent experiments performed in triplicate. **p < 0.01 by Student�s t-test (A) and one-way ANOVA/Turkey post-hoc test (B).
• Data Supplement - Fig. S3. Direct effect of H89 and Ro320432 on 3Hdopamine uptake in TAAR1-DAT cells and wild type (WT) mouse striatal synaptosomes. Cells or synaptosomes were pretreated with 10 μM of H89 or 10 μM Ro320432 alone for 10 min and then washed twice and loaded with 10 nM 3Hdopamine for 5 min. Data shown are values of mean � SEM for three independent experiments performed in triplicate. Neither H89 nor Ro320432 altered 3Hdopamine uptake in TAAR1-DAT cells (A) or in WT mouse striatal synaptosomes (B).
• Data Supplement - Fig. S4. Direct effect of H89 and Ro320432 on 3Hdopamine efflux in TAAR1-DAT cells and wild type (WT) mouse striatal synaptosomes. Cells and synaptosomes were preloaded with 10 nM 3Hdopamine for 20 min and then washed twice and treated with 10μM of H89 or 10 μM Ro320432 for 30 min. Data shown are values of mean � SEM for three independent experiments performed in triplicate. Neither H89 nor Ro320432 altered 3Hdopamine efflux in TAAR1-DAT cells (A) or in WT mouse striatal synaptosomes (B).