Gene expression profile of the SVZ of the postnatal mouse brain

To determine gene expression profiles of neurogenesis, neural stem cell and notch signaling elements in the adult subventricular zone (SVZ) of the WT and CNP-hEGFR mouse, we used SuperArray limited gene array analysis. GEArrayTM expression array systems (cat#OMM404 SuperArray, Bethesda, MD) consisted of spotted cDNA fragments encoding 288 mouse genes related to neurogenesis and neural stem cell differentiation, as well as notch signaling elements. Control sequences (PUC18, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), peptidylpropyl isomerase A (Ppia), and β-actin) were also included. These microarrays were employed to compare SVZ gene expression between WT and CNP-hEGFR mouse SVZ. Total RNA was isolated by Trizol (Invitrogen) and processed for microarray hybridization following the manufacturer’s instructions. Arrays were visualized by autoradiography, and hybridization signals were scanned and analyzed for density in GEArray Expression Analysis Suite 2.0. The normalized value for each gene was calculated by dividing the value of each gene by the average value of the housekeeping genes GAPDH, Ppia, and β-actin. To obtain a gene profile, analysis was performed on three separate RNA samples isolated from different SVZs, with replicate assays conducted on each RNA preparation.