Production of reactive oxygen species, gene expression, and enzymatic activity in quail subjected to acute heat stress1

Abstract

The aim of the present study was to evaluate the effect of acute heat stress on the production of mitochondrial reactive oxygen species (ROS), the gene expression of the avian uncoupling protein (avUCP) and glutathione peroxidase (GPX 7), and the activity of the enzyme GPX in the liver of meat quail. Two groups of 15 meat quail (Coturnix coturnix japonica) that were 23 d of age were initially housed individually in metallic cages. A period of 7 d was provided for the 2 bird groups to adapt to the cages and to a thermoneutral environment at 25°C with 60% relative humidity. At 30 d of age, 15 quail were exposed to a heat stress (HS) treatment of 34°C for 24 h, humidity 60%, whereas control quail (n = 15) were kept at 25°C. To analyze the production of ROS, 4 quail from each treatment group were slaughtered, and their livers were collected for mitochondrial isolation and to measure the subsequent production of ROS by the mitochondria. Additionally, the livers of 6 animals from each treatment group were collected for total RNA extraction. The cDNA was amplified using primers specific for the target genes, and expression was analyzed using the real-time PCR reaction (qRT-PCR). Five animals from each treatment group were slaughtered to analyze glutathione peroxidase (GPx) activity, which was determined by using of hydrogen peroxide (H2O2), and based on measuring the amount nicotinamide adenine dinucleotide phosphate oxidized. A greater amount of mitochondrial ROS was found in HS animals (0.34 vs. 0.22 nm of ROS produced min(-1) · mg(-1) of protein, P < 0.05) for the reactions that contained only rotenone and in the reactions that were performed with rotenone and antimycin (0.31 vs. 0.23 nm of ROS produced min(-1) · mg(-1) of protein, P < 0.05). Concomitantly, the birds that were subjected to acute heat stress and had a greater amount of ROS production expressed less avUCP mRNA [0.75 arbitrary units (AU) vs. 0.87 AU, P < 0.05] and more GPX 7 mRNA (2.37 AU vs. 1.17 AU, P < 0.01). The HS quail displayed significantly greater GPx activity in their hepatocytes (47.8 vs. 39.6 nmol of NADPH oxidized per mg of protein per minute, P < 0.05). Thus, acute heat stress at 34°C for 24 h affects the production of mitochondrial ROS, the expression of avUCP and GPX 7 mRNA, and the activity of the GPx enzyme in the liver of meat quail.