RESUMEN
In industrial poultry, quail production has gained increasing prominence over the years. It is known that the intensification of genetic studies has contributed greatly to this growth, through techniques, such as analysis of gene expression by PCR, for example. This study aimed to evaluate stability and recommend reference genes for quantitative real-time PCR in different tissues from male and female broiler quails. The stability of 10 housekeeping genes (GAPDH, RPL5, MRPS27, MRPS30, TFRC, HMBS, EEF1, LDHA, B2M, and UBC) by means Bestkeeper, NormFinder, GeNorm softwares with ΔCq method. The tissues analyzed were: heart, thigh muscle, brain, and spleen, considering that they are tissues commonly used in nutrigenomic, immunological, and poultry performance research. As expected, the reference genes tested showed varying stability depending on the tissue evaluated. According to the present study, the most stable housekeeping genes were MRPS30, TFRC, and HMBS in heart; MRPS30, EEF1, and HMBS in thigh muscle; B2M, GAPDH, and UBC in brain; and EEF1, LDHA, and HMBS in spleen. Therefore, it is recommended to be used as reference genes for gene expression studies of male and female quails.
Asunto(s)
Pollos , Perfilación de la Expresión Génica , Masculino , Animales , Femenino , Perfilación de la Expresión Génica/métodos , Pollos/genética , Músculo Esquelético/metabolismo , Programas Informáticos , Reacción en Cadena en Tiempo Real de la Polimerasa , Expresión Génica/genéticaRESUMEN
This study was carry out to evaluate mRNA expression of mitochondrial cytochrome c oxidase III in the Pectoralis superficialis muscle of 28-day-old quails fed diets containing 0, 8, and 12% glycerol. Total RNA was extracted (N = 10) and cDNA was amplified using specifics primers for qRT-PCR. Feed efficiency and feed intake were evaluated. COX III mRNA expression in breast muscle was higher in the group fed with 12% glycerol (0.863 AU); no differences were observed in the expression of this gene between the muscle of animals fed diets without glycerol (0.357 AU) and 8% glycerol (0.415 AU). Quails that showed greater COX III mRNA expression also showed the lowest feed efficiency. These results show that there is a difference in COX III mRNA expression in breast muscle of 28-day-old quail fed diets different concentrations of glycerol.
Asunto(s)
Complejo IV de Transporte de Electrones/biosíntesis , Glicerol/administración & dosificación , Músculo Esquelético/efectos de los fármacos , Codorniz/genética , Alimentación Animal , Animales , Dieta , Complejo IV de Transporte de Electrones/genética , Regulación de la Expresión Génica/efectos de los fármacos , Músculo Esquelético/crecimiento & desarrollo , Músculo Esquelético/metabolismo , Codorniz/metabolismo , ARN Mensajero/biosíntesis , ARN Mensajero/genéticaRESUMEN
1. A study was carried out to evaluate the expression of growth hormone (GH), insulin-like growth factor I (IGF-I), mitochondrial adenine nucleotide translocase (ANT), cytochrome oxidase III (COX III) and avian uncoupling protein (avUCP) genes in 14-d-old Japanese quail that were fed different levels of glycerol (0%, 4%, and 12% dietary glycerol) which replaced maize as an energy source. 2. Total RNA was extracted from the breast muscle, and cDNA was amplified using real-time PCR with primers specific to the examined genes. 3. Quail fed the diet with 12% glycerol supplementation presented higher growth hormone (GH) mRNA expression than did those fed 0% glycerol. Supplementation with 12% glycerol negatively influenced IGF-I mRNA expression and reduced ANT mRNA expression in comparison with the treatment with no glycerol. COX III mRNA expression in the pectoralis superficialis muscle was decreased by 26% in quail fed 12% glycerol compared with those fed 0 and 4% glycerol. 4. There was no difference in UCP mRNA expression between quail fed 0 and 4% glycerol; however, UCP expression was reduced (73%) in birds fed the 12% glycerol diet compared with the level in those fed the 4% glycerol diet. 5. The inclusion of 4% glycerol in the diet produced results similar to those of the diet with no glycerol. Based on quail performance and the expression of the GH, IGF-I, ANT, COX III and UCP genes, 4% glycerol can be used in quail feeding without any harmful effects.