Selection of candidate housekeeping genes for normalization in human postmortem brain samples

Ilaria Penna, Serena Vella, Arianna Gigoni, Claudio Russo, Ranieri Cancedda, Aldo Pagano

Research output: Contribution to journalArticle

Abstract

The most frequently used technique to study the expression profile of genes involved in common neurological disorders is quantitative real-time RT-PCR, which allows the indirect detection of very low amounts of selected mRNAs in tissue samples. Expression analysis by RT-qPCR requires an appropriate normalization to the expression level of genes characterized by a stable, constitutive transcription. However, the identification of a gene transcribed at a very stable level is difficult if not impossible, since significant fluctuations of the level of mRNA synthesis often accompanies changes of cell behavior. The aim of this study is to identify the most stable genes in postmortem human brain samples of patients affected by Alzheimer's disease (AD) suitable as reference genes. The experiments analyzed 12 commonly used reference genes in brain samples from eight individuals with AD and seven controls. After a careful analysis of the results calculated by geNorm and NormFinder algorithms, we found that CYC1 and EIF4A2 are the best reference genes. We remark on the importance of the determination of the best reference genes for each sample to be analyzed and suggest a practical combination of reference genes to be used in the analysis of human postmortem samples.

Original languageEnglish
Pages (from-to)5461-5470
Number of pages10
JournalInternational Journal of Molecular Sciences
Volume12
Issue number9
DOIs
Publication statusPublished - Sep 2011

Keywords

  • Alzheimer's disease
  • GeNorm
  • NormFinder
  • Quantitative real-time RT-PCR
  • Reference genes

ASJC Scopus subject areas

  • Computer Science Applications
  • Molecular Biology
  • Catalysis
  • Inorganic Chemistry
  • Spectroscopy
  • Organic Chemistry
  • Physical and Theoretical Chemistry
  • Medicine(all)

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