Mice were treated either during adolescence or adulthood for neurotransmitter receptor and regulator gene expression analysis (Figure1). Messenger RNA was extracted from whole brain using Trizol reagent, followed by purification with RNeasy column (Qiagen, Valencia, CA). The total RNA was converted into first stranded-cDNA using an RT first strand kit (SAB Bioscience Corporation, Frederick, MD). DNAase treatment was included in the kit to ensure the removal of genomic DNA. Quantitative real-time PCR was performed using a Bio-rad MyiQ Single-Color RT PCR Detection System. The Mouse Neurotransmitter Receptors and Regulators RT2 Profiler™ PCR Array, which assesses 84 genes (Supplemental Table 1) was used to perform the reaction (SABioscience Corporation; PAMM-060). The Sybr green DNA Real-Time PCR Mix was purchased from Applied Biosystems (Foster City, CA). The PCR mix was denatured at 95°C for 10 min before the first PCR cycle. The thermal cycle profile was programmed for denaturation for 15 s at 95°C and annealing for 60 s at 60°C. A total of 40 PCR cycles were performed. Data analysis was performed using the ΔCt method using the Data Analysis Template
