Analysis of gene expression in the rat hippocampus using real time PCR reveals high inter-individual variation in mRNA expression levels

被引:35
作者
Alfonso, J
Pollevick, GD
Castensson, A
Jazin, E
Frasch, ACC
机构
[1] Univ Nacl Gen San Martin, Inst Invest Biotecnol, Inst Tecnol Chascomos Consejo Nacl Invest Cient &, Buenos Aires, DF, Argentina
[2] Uppsala Univ, Rudbeck Lab, Dept Genet & Pathol, Med Genet Sect, Uppsala, Sweden
关键词
brain; SYBR Green; differential expression; power calculation;
D O I
10.1002/jnr.10105
中图分类号
Q189 [神经科学];
学科分类号
071006 ;
摘要
In mammals, gene transcription is a step subjected to tight regulation mechanisms. In fact, changes in mRNA levels in the central nervous system (CNS) can account for numerous phenotypic differences in brain function. We performed a high-resolution analysis of mRNA expression levels for 37 genes selected from a normal rat hippocampus cDNA library. mRNA amounts were quantified using a Real Time PCR SYBR Green assay. We found that, in general, individuals from an inbred rat population (n = 20) have shown 2-3 times differences in the basal level of expression of the genes analyzed. Up to several fold differences among individuals were observed for certain genes. These inter-individual differences were obtained after correction for the different amounts of mRNA in each sample. Power calculations were performed to determine the number of individuals required to detect reliable differences in expression levels between a control and an experimental group. These data indicated that, depending on the variability of the candidate gene selected, it was necessary to analyze from five to 135 individuals in each group to detect differences of 50% in the levels of mRNA expression between two groups investigated. The comparison of mRNA abundance from different genes revealed a wide range of expression levels for the 37 genes, showing a 26,000-fold difference between the highest and lowest expressed gene. (C) 2002 Wiley-Liss, Inc.
引用
收藏
页码:225 / 234
页数:10
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