Key Gene Screening for Mammary Fat Deposition in Sheep (Ovis aries) and Construction of a lncRNA-miRNA-mRNA Regulatory Network
ZHANG Yu-Xin1,2, GUO Yan-Yan1,2, LI Yu-Peng1, Guo Xiao-Fei1, ZHANG Xiao-Sheng1, WANG Chao3, RUAN Wei-Bin2, YAO Da-Wei1,*
1 Institute of Animal Science and Veterinary/Tianjin Key Laboratory of Animal Molecular Breeding and Biotechnology/Tianjin Engineering Research Center of Animal Healthy Farming, Tianjin Academy of Agricultural Sciences, Tianjin 300381, China; 2 College of Life Science, Nankai University, Tianjin 300071, China; 3 State Key Laboratory of Farm Animal Biotech Breeding, China Agricultural University, Beijing 100193, China
Abstract：Milk fat synthesis is a critical determinant of the nutritional and economic value of sheep milk, whose post-transcriptional regulatory mechanisms remain incompletely elucidated. This study was conducted to decipher the regulatory role of the competing endogenous RNA (ceRNA) network in milk fat synthesis within the mammary gland of sheep (Ovis aries). Mammary tissue samples were collected from Hu sheep during late pregnancy (14 d prepartum) and peak lactation (14 d postpartum) for whole-transcriptome sequencing. Bioinformatic analyses were performed to construct the ceRNA network and identify target relationships pertinent to lipid synthesis. A total of 1 471 differentially expressed mRNAs were identified between the 2 stages, with 647 upregulated and 824 downregulated during lactation. Among them, suppressor of cytokine signaling 3 (SOCS3), lipin 1 (LPIN1), and stearoyl-CoA desaturase 5 (SCD5) are closely associated with milk fat synthesis. A ceRNA network encompassing 3 mRNAs, 6 miRNAs, and 6 lncRNAs was successfully constructed. Specifically, 2 lncRNAs, MSTRG.15867.1 and XR_006057437.1, were significantly upregulated (P<0.01) in breast tissue at 14 d postpartum, coinciding with the upregulation of their target gene, SOCS3 (P<0.05), forming a putative regulatory axis. Another lncRNA, MSTRG.9360.1, was significantly upregulated (P<0.05) in breast tissue at 14 d postpartum and found to target LPIN1, forming 2 regulatory axes. Furthermore, the MSTRG.15301.43-miR-6715-SCD5 axis was predicted to operate via the ceRNA mechanism. Functional enrichment analysis revealed that the differentially expressed genes were predominantly involved in cell cycle and lipid metabolic pathways, suggesting a coordinated shift from cellular proliferation to active lipid synthesis during lactogenesis. These findings elucidate the involvement of ceRNA networks in regulating milk fat synthesis across different lactation stages in sheep, providing novel insights and potential molecular targets for the breeding of high lactation sheep.
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