Journal
MUSCLE & NERVE
Volume 51, Issue 2, Pages 276-281Publisher
WILEY
DOI: 10.1002/mus.24310
Keywords
denervation; housekeeping gene; immobilization; muscle atrophy; real-time RT-PCR
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Funding
- AIST
- KAKENHI from the Ministry of Education, Culture, Sports, Science, and Technology (MEXT) of Japan [24620015]
- Grants-in-Aid for Scientific Research [24620015] Funding Source: KAKEN
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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. Muscle Nerve 51: 276-281, 2015
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