Maternal Melatonin Supplementation Modulates Placental DNA Methylation and Gene Expression in Nutrient-Restricted Cattle

Authors: RAJPUT, SHIVEELI - Mississippi State University; LITTLEJOHN, BRITTNI - University Of Arkansas; CONTERAS-CORREA, ZULLY - Tarleton State University; EI DAOUS, HALA - Mississippi State University; SIDELINGER, DARCIE - Mississippi State University; KING, E - Mississippi State University; ARICK, MARK - Mississippi State University; LEMLEY, CALEB - Mississippi State University

Submitted to: International Journal of Molecular Sciences
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 11/22/2025
Publication Date: 11/25/2025
Citation: Rajput, S., Littlejohn, B.P., Conteras-Correa, Z.E., Ei Daous, H., Sidelinger, D.R., King, E.H., Arick, M.A., Lemley, C.O. 2025. Maternal Melatonin Supplementation Modulates Placental DNA Methylation and Gene Expression in Nutrient-Restricted Cattle. International Journal of Molecular Sciences. 26 11387. https://doi.org/10.3390/ijms262311387.
DOI: https://doi.org/10.3390/ijms262311387

Interpretive Summary: This study examined how a cow’s nutrition during pregnancy affects the placenta and developing calf, and whether melatonin supplementation can help offset the effects of poor maternal nutrition. Researchers found that nutrient restriction changed important molecular markers in the placenta that regulate how genes are turned on and off, potentially influencing fetal growth and long-term health. Melatonin supplementation altered these molecular signals and appeared to partially counteract some of the negative effects associated with nutrient restriction. Interestingly, male and female fetuses responded differently to melatonin, suggesting that fetal sex influences how the placenta adapts to environmental challenges during pregnancy. In male pregnancies, melatonin affected genes involved in placental function, energy metabolism, and development, while female pregnancies showed widespread molecular changes that may prepare tissues for future developmental responses. These findings suggest that melatonin may help support placental health and fetal development when nutrient availability is limited. Overall, the study provides new insight into how maternal nutrition and melatonin influence the biological programming of offspring before birth and may help identify strategies to improve livestock productivity and offspring health. This research supports the Secretary of Agriculture’s priorities of increasing profitability of farmers.

Technical Abstract: This study investigated the influence of maternal nutrient restriction and dietary melatonin supplementation on DNA methylation and gene expression in bovine placental cotyledons, with a focus on sex-specific changes. On day 160 of gestation, 29 Brangus heifers (bred to a single sire by AI) were subjected to a 2 × 2 factorial design: adequately fed (ADQ-CON, n = 7), nutrient-restricted (RES-CON, n = 7), and adequately fed or nutrient restricted supplemented with 20 mg/d of melatonin (ADQ-MEL, n = 7; RES-MEL, n = 8). Cotyledons were collected at day 240 from 12 female and 17 male conceptuses for Methyl MiniSeq-GWBS and RNA-Seq. In RES-CON vs. ADQ-CON, 93 hypomethylated and 143 hypermethylated DMRs were identified, primarily in exonic, intronic, and promoter regions. Melatonin altered the methylation patterns of male and female cotyledons, respectively, with 203 and 460 DMRs associated with axon guidance, RHOC GTPase cycle, and BDNF signaling pathways. RES-MEL showed higher expression of the UBOX5 gene compared with RES-CON. Moreover, 15 DEGs (5 upregulated and 10 downregulated) were observed in the male vs. female comparison. In melatonin-treated males, PIGX, ATP11C, snoRNA U2-19, ZNF82 genes were upregulated. Thus, melatonin may modulate conceptus growth and development in a sex-specific manner.