Figure 3.

Secondary confirmation of microarray data by in situ hybridization and RNA blot assay. Representative photomicrographs of hippocampal sections from ISH and RNA blot assays using radiolabeled riboprobes are shown, and quantified expression from the indicated cell layers is shown by bar graphs on the right. Results are expressed as a percentage of sham and are the mean ± SEM of four separate animals, each analyzed in duplicate brain sections. Lanes 1-4 in the RNA blots indicate individual samples, each from a separate animal, spotted in duplicate. The housekeeping gene cyclophilin was used to normalize the signal from sham and ECS groups. With the exception of NPY, all ISH images are from acute ECS-treated rats. Upregulation of the neuritin gene shown in A (top) is most evident in the DG granule cell layer after either acute or chronic treatments. A significant increase was also observed in the CA1 pyramidal cell layer after chronic ECS. B, Expression of VEGF in the choroid plexus and induction in the CA1 and CA3 pyramidal cell layers and DG granule cell layer. C, VGF was significantly upregulated in the DG with acute ECS and in the DG and CA3 pyramidal cell layers with chronic ECS. D, Regulation of BDNF was confirmed using the RNA blot assay only. E, FGF-2 shows prominent expression in the CA2 of both sham and ECS groups with maximal induction seen in the DG.