Real-Time PCR Gene Expression Analysis

First-strand cDNA was synthesized by using aliquots of 1 µg of total RNA from cortex and basal ganglia, reverse transcriptase and random primers [7] (link). Specific genes were quantified by real-time PCR using i-Cycler MYIQ system (Bio-Rad, Mississauga, ON). cDNA prepared from total RNA of cultured cells was diluted 1∶1 with sterile water and 5 µl were thereafter used per RT-PCR reaction. Semi-quantitative analysis was performed by monitoring in real time the increase of fluorescence of the SYBR Green dye on a Bio-Rad detection system, as previously reported [57] (link) and expressed as relative fold change (RFC) compared to control. Oligonucleotide primers are provided in Table 1.