Jie Chen1, Runsheng Ruan. 1. Cancer and Ageing Research Group, Institute of Bioengineering and Nanotechnology, 31 Biopolis Way, The Nanos #04-01, Singapore.
Abstract
CONCLUSION: This study suggests that the validation of reference genes is important for the accurate evaluation of differential gene expression in age-related hearing loss (AHL). OBJECTIVES: To identify the most stable reference gene in the auditory cortex and cochlea of young and old rats. Materials and methods. mRNA levels of some common reference genes such as beta-actin, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), ubiquitin C, hypoxanthine phosphoribosyl-transferase and eukaryotic translation elongation factor (EF) were quantified in the auditory cortex and cochlea of young and old rats by quantitative RT-PCR, and then their stability was evaluated using GeNorm and Normfinder software. RESULTS: There were some variations in the expression of reference genes found in the auditory cortex and cochlea of rats, wherein EF and GAPDH were identified as the most stable reference genes in auditory cortex and cochlea, respectively. mRNA levels of Cu/Zn-superoxide dismutase (SOD), Mn-SOD, catalase and glutathione peroxidase in the auditory cortex and cochlea do not vary significantly in young and old rats, when they are normalized with EF or GAPDH, the valid reference genes, respectively. However, Cu/Zn-SOD and catalase levels differ significantly in the auditory cortex when they are normalized with beta-actin, an invalid reference gene, resulting in the misinterpretation of the target gene expression levels.
CONCLUSION: This study suggests that the validation of reference genes is important for the accurate evaluation of differential gene expression in age-related hearing loss (AHL). OBJECTIVES: To identify the most stable reference gene in the auditory cortex and cochlea of young and old rats. Materials and methods. mRNA levels of some common reference genes such as beta-actin, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), ubiquitin C, hypoxanthine phosphoribosyl-transferase and eukaryotic translation elongation factor (EF) were quantified in the auditory cortex and cochlea of young and old rats by quantitative RT-PCR, and then their stability was evaluated using GeNorm and Normfinder software. RESULTS: There were some variations in the expression of reference genes found in the auditory cortex and cochlea of rats, wherein EF and GAPDH were identified as the most stable reference genes in auditory cortex and cochlea, respectively. mRNA levels of Cu/Zn-superoxide dismutase (SOD), Mn-SOD, catalase and glutathione peroxidase in the auditory cortex and cochlea do not vary significantly in young and old rats, when they are normalized with EF or GAPDH, the valid reference genes, respectively. However, Cu/Zn-SOD and catalase levels differ significantly in the auditory cortex when they are normalized with beta-actin, an invalid reference gene, resulting in the misinterpretation of the target gene expression levels.
Authors: Chiemi Tanaka; Donald E Coling; Senthilvelan Manohar; Guang-Di Chen; Bo Hua Hu; Richard Salvi; Donald Henderson Journal: Neurobiol Aging Date: 2012-02-01 Impact factor: 4.673
Authors: Ana Carolina de Moura; Virgínia Meneghini Lazzari; Grasiela Agnes; Silvana Almeida; Márcia Giovenardi; Ana Beatriz Gorini da Veiga Journal: Einstein (Sao Paulo) Date: 2014-09
Authors: Pedro Melgar-Rojas; Juan Carlos Alvarado; Verónica Fuentes-Santamaría; María Cruz Gabaldón-Ull; José M Juiz Journal: PLoS One Date: 2015-09-14 Impact factor: 3.240