Highest trkB mRNA expression in the entorhinal cortex among hippocampal subregions in the adult rat: contrasting pattern with BDNF mRNA expression

Spatiotemporal Dynamics of Brain-Derived Neurotrophic Factor mRNA Induction in the Vestibulo-Olivary Network during Vestibular Compensation

“…In this method, the quantity of the target gene was normalized with that of the internal standard gene, and this normalization canceled out variations that occurred during PCR amplification. We have confirmed previously that the quantifiability and reproducibility of this method are comparable with those of RNase protection assays and Northern blotting analysis (Tokuyama et al, 1998Okuno et al, 1999). Our PCR-based quantification method thus allows for reliable and efficient analysis of multiple genes when RNA amounts are limited and so is most appropriate for the quantitative evaluation of changes in mRNA expression in the small brainstem regions.…”

“…The brains were cut coronally into 150-m-thick sections with a cryostat. Each section was placed in a dish containing chilled PBS just before tissue dissection as described previously (Tokuyama et al, 1998. The tissue dissection was performed under a stereomicroscope on the basis of the borders in an atlas of the brainstem described by Paxinos et al (1999).…”

“…All tissues were separately excised from the ipsilateral and contralateral sides to the lesion, collected, frozen immediately on dry ice, and stored at Ϫ80°C until RNA extraction (Tokuyama et al, 1998). For each brain area, we pooled excised tissues from six animals to obtain sufficient tissue for mRNA quantification.…”

“…E xpression levels of mRNAs were quantified by using an RT-PCR coamplification method as described previously (Tokuyama et al, 1998Okuno et al, 1999). Specific primers for PCR coamplification were designed based on the published sequences of rat BDN F (GenBank accession number M61175), trkB (GenBank accession number M55291), c-fos (GenBank accession number X06769), ␣-tubulin (GenBank accession number V10227), and hypoxanthineguanine phosphoribosyltransferase (hprt) (GenBank accession number M63983) genes.…”

“…The procedures of reverse transcription and PCR coamplification were essentially the same as described previously (Tokuyama et al, 1998;Okuno et al, 1999). In the present study, the hprt gene was used as an internal standard gene .…”