Muscle & nerve · 2014
Impact of denervation-induced muscle atrophy on housekeeping gene expression in mice.
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Nakao Reiko, Yamamoto Saori, Yasumoto Yuki, Kadota Koji, Oishi Katsutaka
Аннотация
INTRODUCTION: 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.
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.
Предметные рубрики MeSH
Analysis of VarianceAnimalsDenervationDisease Models, AnimalFunctional LateralityGene Expression RegulationGenes, EssentialGlyceraldehyde-3-Phosphate DehydrogenasesHypoxanthine PhosphoribosyltransferaseMaleMiceMice, Inbred C57BLMuscle, SkeletalMuscular AtrophyRNA, MessengerSciatic NeuropathyTATA-Box Binding ProteinTime Factors