Characterization of body composition and liver epigenetic markers during periods of negative energy balance and subsequent compensatory growth in postpubertal beef bulls

This study aimed to characterize the effects of dietary restriction and subsequent re-alimentation on body composition and hepatic gene expression of epigenetic markers of DNA methylation, RNA m6A methylation, and histone acetylation in the liver of postpubertal beef bulls. Twelve Angus x Hereford crossbred bulls (n = 6, 23 +/- 0.55 mo [young bulls], 558 +/- 6.1 kg; and n = 6, 47 +/- 1.2 mo [mature bulls], 740 +/- 30.5 kg) were submitted to two dietary regimes per offering of the same hay: low plane of nutrition (90 d) and compensatory growth (90 d). Each animal acted as its own control and were fed Beardless wheat (Triticum aestivum) hay and mineral mix during the trial. Statistical analyses were performed using SAS 9.4 following a pre-post repeated measures design. Bulls in negative energy balance (NEB) decreased (P < 0.001) empty body weight (EBW; 23.1% [-139.1 kg]), empty body fat (EBF; 39.8% [-85.4 kg]), and empty body protein (EBP; 14.9% [-13.5 kg]) and fully recovered at the end of the trial. Body fat accounted for 77.1% of daily changes in body energy status, whereas body protein accounted for only 22.9% (P < 0.001). Relative abundance of epigenetic markers transcripts was analyzed via qPCR. Bulls at NEB tended (P <= 0.097) to increase gene expression of epigenetic markers of RNA m6A methylation (METTL14, VIRMA, and WTAP) and increased (P <= 0.050) the gene expression of epigenetic markers of DNA methylation (DNMT3A) and histone-acetylation (SIRT3 and SIRT7). Young bulls had a tendency (P <= 0.072) of higher RNA m6A methylation, VIRMA, and WTAP than mature bulls. Effect of diet x age interaction was not detected (P >= 0.137) for METTL14, VIRMA, WTAP, DNMT3A, SIRT3, or SIRT7. Younger bulls tended to have greater RNA m6A methylation levels than mature bulls, indicating that, while contemporaneously fed the same diet during periods of undernourishment followed by compensatory growth, age has an impact on this epigenetic mechanism. In conclusion, metabolic status seems to carry a greater impact on regulating bovine hepatic epigenetic mechanisms that modulate gene transcription, such as DNA methylation and histone acetylation, than on epigenetic mechanisms that regulate gene translation, such as RNA m6A methylation. During periods of undernourishment followed by compensatory growth, body fat pools appear to change more dynamically and are easily detected having a greater impact on epigenetic markers that modulate hepatic gene transcription rather than translation. Lay Summary Epigenetics refers to heritable modifications that regulate gene expression without altering DNA sequence, hence, acting on top of the genes. Epigenetic markers change in response to stressors such as environmental factors, nutritional challenges, among other overlooked players that altogether could drastically impair animal performance. During periods of undernourishment followed by fast weight gain, dynamic changes in body composition, especially fat, appear to trigger an increased action of such physiological markers that modulate hepatic gene expression. Findings of this study unveil epigenetic metabolic pathways that deserve further investigation for proper quantification of potential consequences of metabolic stress on the liver of bovines that suffer significant loss of body weight followed by recovery. The alterations at the molecular level shown in this study provide a picture of silent metabolic changes that have not been detected previously in liver metabolism studies of cattle. Therefore, the impact of nutritional management and metabolic stress may be greater than previously expected and differently controlled than previously assumed. Molecular analyses unveil previously undetected changes on the liver of bulls in response to diet and body weight fluctuation.