Abstract
We investigated the nutritional effects on carcass traits, gene expression and DNA methylation in a three generation Large White pig feeding experiment. A group of experimental (E) F0 boars were fed a standard diet supplemented with high amounts of methylating micronutrients whereas a control group (C) of F0 boars received a standard diet. These differentially fed F0 boars sired F1 boars which then sired 60 F2 pigs. Carcass traits were compared between 36 F2 descendants of E F0 boars and 24 F2 descendants of C F0 boars. The two F2 offspring groups differed with respect to backfat percentage (P = 0.03) and tended to differ with respect to adipose tissue (P = 0.09), fat thickness at the 10th rib (P = 0.08) and at the croup (P = 0.09) as well as percentages of shoulder (P = 0.07). Offspring from the experimental F0 boars had a higher percentage of shoulder and were leaner compared to the control group. Gene expression profiles showed significant twofold differences in mRNA level between 8 C F2 offspring and 8 E F2 offspring for 79, 64 and 53 genes for muscle, liver and kidney RNA, respectively. We found that in liver and muscle respective pathways of lipid metabolism and metabolic pathway were over-represented for the differentially expressed genes between these groups. A DNA methylation analysis in promoters of differentially expressed genes indicated a significant difference in DNA methylation at the IYD gene. If these responses on carcass traits, gene expression and DNA methylation withstand verification and can indeed be attributed to transgenerational epigenetic inheritance, it would open up pioneering application in pork production and would have implications for human health.
Highlights
There is a growing body of evidence that environmental effects including nutrition affect the epigenetic code in mammals and that such induced modifications are transmitted to generations [1,2,3]
Three Generation Pig Feeding Experiment The micronutrients added to the diet E were selected based on the one-carbon metabolism involved in DNA methylation
We found significant differences in DNA methylation at the IYD promoter between C and E F2 offspring which is associated with gene expression
Summary
There is a growing body of evidence that environmental effects including nutrition affect the epigenetic code in mammals and that such induced modifications are transmitted to generations [1,2,3]. It was observed that DNA methylation in liver was modestly changed at various loci including a likely enhancer for the lipid regulator Ppara [12] In these examples the transmission of epigenetic modifications and their associated phenotypes were demonstrated, the unequivocal segregation of an induced epimutation between generations needs to be proven in order to claim true transgenerational epigenetic inheritance. We hypothesized that environmental perturbations such as dietary modifications affects the epigenetic code in pigs which is transmitted up to two generations later For this purpose F0 boars were fed a diet enriched with methyl donors or a control diet to investigate heritable epigenetic effects in F2 offspring phenotypes, which would be truly transgenerational in nature. We further present a pathway analysis of microarray gene expression data and studied DNA methylation in promoter regions of differentially expressed genes
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