Morphological adaptation of sheep's rumen epithelium to high-grain diet entails alteration in the expression of genes involved in cell cycle regulation, cell proliferation and apoptosis

Abstract

Background: The objectives of this study were to characterize changes in the relative mRNA expression of candidate genes and proteins involved in cell cycle regulation, cell proliferation and apoptosis in the ruminal epithelium (RE) of sheep during high-grain (HG) diet adaptation. Results: Twenty sheep were assigned to four groups with five animals each. These animals were assigned to different periods of HG diet (containing 40% forage and 60% concentrate mix) feeding. The HG groups received an HG diet for 7 (G7, n = 5), 14 (G14, n = 5) and 28 d (G28, n = 5), respectively. In contrast, the control group (CON, n = 5) was fed the forage-based diet for 28 d. The results showed that HG feeding linearly decreased (P < 0.001) the ruminal pH, and increased the concentrations of ruminal total volatile fatty acid (linear, P = 0.001), butyrate (linear, P < 0.001), valerate (quadratic P = 0.029) and the level of IGF-1 (quadratic, P = 0.043) in plasma. The length (quadratic, P = 0.004), width (cubic, P = 0.015) and surface of the ruminal papillae (linear, P = 0.003) were all enlarged after 14 d of HG diet feeding. HG feeding cubically increased the number of cell layers forming the stratum corneum (SC, P < 0.001) and the thickness of the SC (P < 0.001) and stratum basale (P < 0.001). The proportion of basal layer cells in the RE decreased (linear, P < 0.001) in the G0/G1-phase, but it increased linearly (P = 0.006) in the S-phase and cubically (P = 0.004) in the G2/M-phases. The proportion of apoptosis cells in G7, G14 and G28 was reduced compared to the CON (quadratic, P < 0.001). HG diet feeding linearly decreased the mRNA expression of Cyclin E1 (P = 0.021) and CDK-2 (P = 0.001) and (P = 0.027) the protein expression of Cyclin E1. Feeding an HG diet linearly increased the mRNA expression of genes IGFBP-2 (P = 0.034) and IGFBP 5 (P < 0.009), while linearly decreasing (P < 0.001) the IGFBP 3 expression. The expression of cell apoptosis gene Caspase 8 decreased (quadratic, P = 0.012), while Bad mRNA expression tended to decrease (cubic, P = 0.053) after HG feeding. Conclusions: These results demonstrated sequential changes in rumen papillae size, cell cycle regulation and the genes involved in proliferation and apoptosis as time elapsed in feeding a high-grain diet to sheep.