Decreased beta-actin mRNA expression in hyperglycemic focal cerebral ischemia in the rat

Neurosci Lett. 2004 Mar 11;357(3):211-4. doi: 10.1016/j.neulet.2003.12.081.

Abstract

beta-Actin is often used as a housekeeping gene when performing reverse transcription-polymerase chain reaction (RT-PCR) analysis for cerebral ischemia models. In the present study, we tested two different control genes used for RT-PCR experiments, beta-actin and porphobilinogen deaminase (PBG-D), in a rat model of focal cerebral ischemia under normo- or hyperglycemic conditions. A three-vessel occlusion model with permanent middle cerebral artery occlusion was used in the rat. beta-Actin mRNA expression was decreased in hyperglycemic ischemic rats compared to normoglycemic ischemic animals 3 h post-ischemia. beta-Actin protein content was unchanged. As for PBG-D, its mRNA expression remained constant throughout the groups. Our data thus show that, following focal cerebral ischemia in hyperglycemic conditions, beta-actin is an unsuitable housekeeping gene whereas PBG-D is more appropriate. This study clearly demonstrates the importance of selecting a stable housekeeping gene when performing RT-PCR experiments.

Publication types

  • Comparative Study

MeSH terms

  • Actins / genetics
  • Actins / metabolism*
  • Animals
  • Blotting, Northern / methods
  • Brain Ischemia / complications
  • Brain Ischemia / genetics
  • Brain Ischemia / metabolism*
  • Disease Models, Animal
  • Food Deprivation
  • Gene Expression Regulation / physiology*
  • Hydroxymethylbilane Synthase / genetics
  • Hyperglycemia / complications
  • Hyperglycemia / genetics
  • Hyperglycemia / metabolism*
  • Male
  • Nitric Oxide Synthase / genetics
  • Nitric Oxide Synthase / metabolism
  • Nitric Oxide Synthase Type II
  • RNA, Messenger / metabolism
  • Rats
  • Rats, Sprague-Dawley
  • Reverse Transcriptase Polymerase Chain Reaction / methods
  • Time Factors

Substances

  • Actins
  • RNA, Messenger
  • Nitric Oxide Synthase
  • Nitric Oxide Synthase Type II
  • Nos2 protein, rat
  • Hydroxymethylbilane Synthase