Effects of aging, dietary restriction and glucocorticoid treatment on housekeeping gene expression in rat cortex and hippocampus-evaluation by real time RT-PCR
作者： N. Tanic , M. Perovic , A. Mladenovic , S. Ruzdijic , S. Kanazir
DOI: 10.1007/S12031-007-0006-7
关键词: Cerebral cortex 、 Peptidylprolyl isomerase 、 TaqMan 、 Housekeeping gene 、 Gene expression 、 Molecular biology 、 Real-time polymerase chain reaction 、 Gene expression profiling 、 Reference genes 、 Biology
摘要: Accurate normalization is the prerequisite for obtaining reliable results in quantification of gene expression. Using TaqMan Real Time RT-PCR, we carried out an extensive evaluation five most commonly used endogenous controls, gapdh, beta-actin, 18S rRNA, hprt and cypB, their presumed stability expression, rat cortex hippocampus, during aging, under dietary restriction dexamethasone treatment. Valid reference genes (HKGs) were identified using GeNorm NormFinder software packages by direct comparison Ct values. Analysis revealed gapdh beta-actin as stable HKGs all treatments analyzed, combined or separately, cortex, while hippocampus gapdh/hprt beta-actin/hprt are combination choice single effects restriction/dexamethasone, respectively. All significantly influenced expression rRNA cypB both structures. In addition, factor, calculated GeNorm, to compare alpha-syn cortex. Our demonstrate importance right HKG suggest appropriate control be RT-PCR analysis mRNA selected experimental paradigms.
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