title: Impact of denervation-induced muscle atrophy on housekeeping gene expression in mice.
abstract: BACKGROUND
  Immobilization induced by experimental denervation leads to rapid and progressive alterations in structural and biochemical properties of skeletal muscle. Real-time reverse transcription-polymerase chain reaction (RT-PCR) is a popular method of elucidating the molecular mechanisms involved in muscle atrophy. Identification of suitable reference genes that are not affected by experimental conditions is a critical step in accurate normalization of real-time RT-PCR.
  
  
  METHODS
  We investigated the impact of denervation-induced muscle atrophy for 2 weeks on the expression of common housekeeping genes.
  
  
  RESULTS
  Denervation differentially affected the expression levels of these genes. RefFinder software identified TATA box binding protein (Tbp) as the most stable gene and showed that the stability of glyceraldehyde-3-phosphate dehydrogenase (Gapdh) and hypoxanthine guanine phosphoribosyl transferase (Hprt) genes was low, even though they are widely used for normalization.
  
  
  CONCLUSIONS
  The appropriate reference gene for normalization of genes of interest in denervated muscle is Tbp.
authors.affiliation: Biological Clock Research Group, Biomedical Research Institute, National Institute of Advanced Industrial Science and Technology, Central 6, 1-1-1 Higashi, Tsukuba, Ibaraki, 305-8566, Japan.
authors.forename: Reiko
authors.initials: R
authors.lastname: Nakao
authors.forename: Saori
authors.initials: S
authors.lastname: Yamamoto
authors.forename: Yuki
authors.initials: Y
authors.lastname: Yasumoto
authors.forename: Koji
authors.initials: K
authors.lastname: Kadota
authors.forename: Katsutaka
authors.initials: K
authors.lastname: Oishi
mesh_terms: D000704:Analysis of Variance; D000818:Animals; D003714:Denervation / Q000009:adverse effects; D004195:Disease Models, Animal; D007839:Functional Laterality; D005786:Gene Expression Regulation / Q000502:physiology*; D020043:Genes, Essential / Q000235:genetics*; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000235:genetics / Q000378:metabolism; D007041:Hypoxanthine Phosphoribosyltransferase / Q000235:genetics / Q000378:metabolism; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D018482:Muscle, Skeletal / Q000378:metabolism; D009133:Muscular Atrophy / Q000209:etiology* / Q000378:metabolism*; D012333:RNA, Messenger / Q000378:metabolism; D020426:Sciatic Neuropathy / Q000150:complications*; D035181:TATA-Box Binding Protein / Q000235:genetics / Q000378:metabolism*; D013997:Time Factors
chemical_list: D012333:RNA, Messenger; D035181:TATA-Box Binding Protein; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases; D007041:Hypoxanthine Phosphoribosyltransferase
keywords: denervation; housekeeping gene; immobilization; muscle atrophy; real-time RT-PCR