去神经支配诱导的肌肉萎缩对小鼠管家基因表达的影响。
Impact of denervation-induced muscle atrophy on housekeeping gene expression in mice.
作者信息
Nakao Reiko, Yamamoto Saori, Yasumoto Yuki, Kadota Koji, Oishi Katsutaka
机构信息
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.
出版信息
Muscle Nerve. 2015 Feb;51(2):276-81. doi: 10.1002/mus.24310. Epub 2014 Dec 23.
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.
引言
实验性去神经支配诱导的固定会导致骨骼肌结构和生化特性迅速且渐进性改变。实时逆转录聚合酶链反应（RT-PCR）是阐明肌肉萎缩相关分子机制的常用方法。鉴定不受实验条件影响的合适内参基因是实时RT-PCR准确定量的关键步骤。
方法
我们研究了去神经支配诱导的肌肉萎缩2周对常见管家基因表达的影响。
结果
去神经支配对这些基因的表达水平有不同影响。RefFinder软件将TATA盒结合蛋白（Tbp）鉴定为最稳定的基因，并表明甘油醛-3-磷酸脱氢酶（Gapdh）和次黄嘌呤鸟嘌呤磷酸核糖基转移酶（Hprt）基因的稳定性较低，尽管它们被广泛用于定量分析。
结论
去神经支配肌肉中感兴趣基因定量分析的合适内参基因是Tbp。
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