Quantitative assay of 5-HT1A receptor gene expression in the brain

5-HT1A receptors are involved in the regulation of various behaviors and the mechanism of action of anxiolytics and antidepressants. It is rather difficult to study the expression of the 5-HT1A receptor gene in the brain because of the low concentration of its mRNA. A method developed for quantitating the level of 5-HT1A receptor gene expression in brain structures involves estimation of the copy number for contaminant genomic DNA, the cDNA of the glyceraldehyde 3-phosphate dehydrogenase (GAPDH) gene (a housekeeping gene), and the 5-HT1A receptor gene cDNA in a cDNA preparation. To estimate the GAPDH and 5-HT1A receptor cDNA copy numbers, the fluorescent intensity of the corresponding PCR products is calibrated using genomic DNA standards of known concentrations. The expression of the 5-HT1A receptor gene is corrected for the content of contaminant genomic DNA and presented as a 5-HT1A receptor cDNA copy number per 100 copies of the GAPDH cDNA. The method was used to demonstrate for the first time that expression of the 5-HT1A receptor gene is increased in the frontal cortex and the amygdala of mice knocked-out in the monoamine oxidase A gene. © Pleiades Publishing, Inc., 2006.