Impact of denervation-induced muscle atrophy on housekeeping gene expression in mice.

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.