Omics & gene networks

Network Preservation Analysis Reveals Intergenerational Effects of Prepubertal Protein Restriction in Replacement Ewes on Offspring Metabolic Tissues

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Abstract

Adequate dietary protein intake is essential for animals to reach their genetic potential during growth. However, the increasing cost and limited availability of protein sources pose significant challenges for sustainable livestock production. In growing animals, insufficient protein intake can impair nutrient metabolism, reduce insulin sensitivity, and compromise immune function, with effects that may persist beyond the restriction period. Moreover, growing evidence suggests that maternal nutrition may induce epigenetic modifications with intergenerational effects on metabolic tissues such as liver, muscle, and adipose tissue. This study aimed to investigate how a prepubertal dietary protein restriction influences offspring transcriptomic profiles by evaluating the stability of gene co-expression networks across liver, muscle, and adipose tissues and identifying biological pathways associated with potential epigenetic reprogramming effects. To address these objectives, transcriptomic analyses were performed on 16 Spanish Assaf suckling lambs born to primiparous ewes that had previously been subjected to a prepubertal dietary protein restriction. These replacement ewes received either a standard diet (Control, n = 8) or a protein-restricted diet between 3 and 5 months of age (MNPR, n = 8), after which all were fed identically until their first lambing. The lambs were reared on artificial milk replacer and slaughtered at the commercial weight of approximately 11 kg. At slaughter, liver, muscle, and tail fat samples were collected for subsequent RNA-Seq analysis. Weighted Gene Co-expression Network Analysis (WGCNA) was applied to RNA-Seq data to construct tissue-specific networks for each experimental group. Preservation analysis was then performed to evaluate the stability of co-expression patterns (modules), using the Control network as the reference and the MNPR network as the test, based on Zsummary and medianRank statistics. This approach enabled the identification of non-preserved gene co-expression networks between Control and MNPR offspring, reflecting tissue-specific disruptions in network stability and potential epigenetic reprogramming effects induced by the maternal protein restriction. Two modules in the liver, four in muscle, and two in tail fat exhibited the most unstable co-expression patterns within the MNPR network. Functional enrichment analysis performed on these modules highlighted processes related to cellular energy homeostasis, enzyme activity, and signal transduction, with mitochondria as a central hub connecting these pathways. These findings suggest that dietary protein restriction during puberty in ewes may lead to mitochondrial dysfunction across tissues in offspring, potentially altering energy metabolism and increasing susceptibility to metabolic disorders. Collectively, these insights advance our understanding of how early-life maternal nutrition shapes offspring metabolism and may guide strategies to optimize developmental programming and production efficiency in livestock.

Keywords: 2026

How to Cite:

Vrcan, M., Suárez-Vega, A., Fonseca, P., Pelayo, R., Arranz, J. & Gutiérrez-Gil, B., (2026) “Network Preservation Analysis Reveals Intergenerational Effects of Prepubertal Protein Restriction in Replacement Ewes on Offspring Metabolic Tissues”, World Congress on Genetics Applied to Livestock Production Digital Archive 2026(1): 2284000. doi: https://doi.org/10.31274/wcgalp.23544

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