Quantitative RT-PCR assay of 5-HT1A and 5-HT2A serotonin receptor mRNAs using genomic DNA as an external standard

Brain serotonin 5-HT1A and 5-HT2A receptors have been implicated in both normal and pathological behavior, and in the action of anxiolytic and antidepressant drugs. In this study, detailed description and verification of a new RT-PCR technique to quantify the number of copies of 5-HT1A and 5-HT2A receptor mRNAs in the brain is presented. The number of copies of β-actin and 5-HT1A or 5-HT2A receptor mRNAs in rat brain samples was evaluated with respect to the genomic DNA solution as the external exogenous standard. The expression of 5-HT receptors was calculated as the number of receptor mRNA copies per 100 copies of corresponding β-actin mRNA. This presented technique is reliable, simple and can be easily set up in any neurobiological laboratory.