A comparison between different reference genes for expression studies in human hippocampal tissue

J Neurosci Methods. 2012 Jun 30;208(1):44-7. doi: 10.1016/j.jneumeth.2012.04.020. Epub 2012 May 3.

Abstract

The reliability of gene expression studies by mRNA quantification is highly dependent upon several experimental procedures, including the choice of reference genes used for data normalization. In order to contribute to gene expression studies in mesial temporal lobe epilepsy (MTLE) we used microarray data, followed by real time quantitative PCR validation of selected housekeeping genes, to determine the most appropriate reference genes to be used in human hippocampal tissue gene expression studies. Our results unequivocally showed a significant impact of the reference gene chosen for normalization on the overall results of expression studies, clearly demonstrating the importance of adequate validation using stable reference genes. In addition, we found that HPRT, NSE, SDHA and SYP are suitable genes to be used as reference for normalization in expression studies of hippocampal tissue obtained from patients with MTLE.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Epilepsy, Temporal Lobe / metabolism*
  • Gene Expression Profiling / methods
  • Gene Expression Profiling / standards*
  • Hippocampus / metabolism*
  • Humans
  • Nerve Tissue Proteins / metabolism*
  • Protein Array Analysis / methods
  • Protein Array Analysis / standards*
  • Real-Time Polymerase Chain Reaction / methods
  • Real-Time Polymerase Chain Reaction / standards*
  • Reference Values
  • Reproducibility of Results
  • Sensitivity and Specificity

Substances

  • Nerve Tissue Proteins