Fig 1.
The treatments are described as follows: CON, chicks hatched from control eggs without in ovo injection and incubated at standard temperature; CON+HS, chicks hatched from control eggs without in ovo injection, incubated at standard temperature but exposed to HS; G10+HS, chicks hatched from eggs injected at 17.5 days of incubation with 0.6mL of 10% GABA dissolved in distilled water and exposed to HS; TM+HS, chicks hatched from thermally manipulated eggs exposed to 39.6°C for 6 h daily from ED 10 to 18 and exposed to HS; G10+TM+HS, chicks, hatched from eggs that received both previous treatments during incubation, and exposed to HS.
Table 1.
Oligonucleotide primer sequences for RT-qPCR.
Table 2.
Effects of in ovo feeding of GABA and embryonic thermal manipulation on absolute and relative organs weight in broiler chickens exposed to cyclic HS.
Table 3.
Effects of in ovo feeding of GABA and embryonic thermal manipulation on plasma biochemical parameters in broiler chickens exposed to cyclic HS.
Fig 2.
Effects of in ovo feeding of GABA and embryonic thermal manipulation on relative mRNA expression of hepatic SOD (A), CAT (B), GPx1 (C), NOX1 (D), NOX4 (E), and NRF2 (F) in broiler chickens exposed to cyclic HS. The treatments are described as follows: CON, chicks hatched from control eggs without in ovo injection and incubated at standard temperature; CON+HS, chicks hatched from control eggs without in ovo injection, incubated at standard temperature but exposed to HS; G10+HS, chicks hatched from eggs injected at 17.5 days of incubation with 0.6mL of 10% GABA dissolved in distilled water and exposed to HS; TM+HS, chicks hatched from thermally manipulated eggs exposed to 39.6°C for 6 h daily from ED 10 to 18 and exposed to HS; G10+TM+HS, chicks hatched from eggs that received both previous treatments during incubation and exposed to HS. Data show mean ± SEM (n = 5). Results of the contrast analysis are indicated in the graph. Abbreviations: CAT, catalase; GPx1, glutathione peroxidase 1; NOX1, nicotinamide adenine dinucleotide phosphate oxidase 1; NOX4, nicotinamide adenine dinucleotide phosphate oxidase 4; NRF2: Nuclear factor erythroid 2-related factor 2; SOD, superoxide dismutase.
Fig 3.
Effects of in ovo feeding of GABA and embryonic thermal manipulation on relative mRNA expression of hepatic ACC (A), FAS (B), EXFABP (C), and PPAR-γ (D) in broiler chickens exposed to cyclic HS. The treatments are described as follows: CON, chicks hatched from control eggs without in ovo injection and incubated at standard temperature; CON+HS, chicks hatched from control eggs without in ovo injection, incubated at standard temperature but exposed to HS; G10+HS, chicks hatched from eggs injected at 17.5 days of incubation with 0.6mL of 10% GABA dissolved in distilled water and exposed to HS; TM+HS, chicks hatched from thermally manipulated eggs exposed to 39.6°C for 6 h daily from ED 10 to 18 and exposed to HS; G10+TM+HS, chicks hatched from eggs that received both previous treatments during incubation and exposed to HS. Data show mean ± SEM (n = 5). Results of the contrast analysis are indicated in the graph. Abbreviations: ACC, acetyl-CoA carboxylase; EXFABP: Extracellular fatty acid-binding protein; FAS, fatty acid synthase; PPAR-γ: Peroxisome proliferator-activated receptor-gamma.
Fig 4.
Effects of in ovo feeding of GABA and embryonic thermal manipulation on relative mRNA expression of hepatic HSP70 (A), and HSP90 (B) in broiler chickens exposed to cyclic HS. The treatments are described as follows: CON, chicks hatched from control eggs without in ovo injection and incubated at standard temperature; CON+HS, chicks hatched from control eggs without in ovo injection, incubated at standard temperature but exposed to HS; G10+HS, chicks hatched from eggs injected at 17.5 days of incubation with 0.6mL of 10% GABA dissolved in distilled water and exposed to HS; TM+HS, chicks hatched from thermally manipulated eggs exposed to 39.6°C for 6 h daily from ED 10 to 18 and exposed to HS; G10+TM+HS, chicks hatched from eggs that received both previous treatments during incubation and exposed to HS. Data show mean ± SEM (n = 5). Results of the contrast analysis are indicated in the graph. Abbreviations: HSP70, heat-shock protein 70; HSP90, heat shock protein 90.
Fig 5.
Boxplots showing the assessment of the effects of in ovo feeding of GABA and embryonic thermal manipulation on relative mRNA expression of the set of antioxidant-related genes (A), fatty acid-related genes (B), and heat shock proteins (C) using MANOVA. The treatments are described as follows: CON, chicks hatched from control eggs without in ovo injection and incubated at standard temperature; CON+HS, chicks hatched from control eggs without in ovo injection, incubated at standard temperature but exposed to HS; G10+HS, chicks hatched from eggs injected at 17.5 days of incubation with 0.6mL of 10% GABA dissolved in distilled water and exposed to HS; TM+HS, chicks hatched from thermally manipulated eggs exposed to 39.6°C for 6 h daily from ED 10 to 18 and exposed to HS; G10+TM+HS, chicks hatched from eggs that received both previous treatments during incubation and exposed to HS.
Table 4.
Results of MANOVA and multivariate planned contrasts on sets of hepatic genes expression in broiler chickens exposed to HS.
Fig 6.
Pearson correlation heat map between the relative mRNA levels of the genes studied.
The red color indicates a positive correlation, the blue color indicates a negative correlation and the white color indicates no correlation. Pearson r values were calculated using the “CORR procedure” of the SAS software version 9.4 (SAS Institute Inc., 2009). Abbreviations: ACC, acetyl-CoA carboxylase; CAT, catalase; EXFABP, extracellular fatty acid-binding protein; FAS, fatty acid synthase; GPx1, HSP70, heat-shock protein 70; HSP90, heat shock protein 90; glutathione peroxidase 1; NOX1, nicotinamide adenine dinucleotide phosphate oxidase 1; NOX4, nicotinamide adenine dinucleotide phosphate oxidase 4; NRF2, nuclear factor erythroid 2-related factor 2; PPAR-γ, peroxisome proliferator-activated receptor-gamma; SOD, superoxide dismutase. + Correlation is significant at the P < 0.1. * Correlation is significant at the 0.05 level. ** Correlation is significant at the 0.01 level. *** Correlation is significant at the 0.001 level.