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Goat milk in some cases is less allergenic than cow milk, therefore, more people drink goat milk in the world, so it is necessary for us to improve the yield and quality of goat milk. Previous studies have shown that some genes are closely related to lactation. Ovarian cancer G protein-coupled 1 (OGR1) is a G protein-coupled receptor discovered recently. OGR1 is widely found in various tissues of organisms and is involved in cell skeleton reorganization, carcinogenesis, cell proliferation, and apoptosis by regulating multiple signaling pathways in cells. However, the modulating effect of OGR1 in lactation is still unknown. Therefore, the objective of this study is to investigate the function of OGR1 in goat mammary epithelial cells (GMECs). Flow cytometry, CCK8, EDU, enzyme-linked immunosorbent assay, and triglyceride test kit assays were performed and we found that OGR1 regulated Bcl-2/Bax ratio, Fas protein expression as well as the phosphorylation of AKT and mammalian target of rapamycin (mTOR). si-OGR1 could enhance the proliferation of GMECs by promoting G1/S phase progression and the synthesis of β-casein and triglyceride. By contrast, OGR1 repressed GMECs proliferation and down-regulated the synthesis of β-casein and triglyceride by blocking the PI3K/AKT/mTOR signaling pathway in GMECs.

Background: Protein phosphorylation plays an important role in lactation. Differentially modified phosphorylation sites and phosphorylated proteins between peak lactation (PL, 90 days postpartum) and late lactation (LL, 280 days postpartum) were investigated using an integrated approach, namely, liquid chromatography with tandem mass spectrometry (LC-MS/MS) and tandem mass tag (TMT) labeling, to determine the molecular changes in the mammary tissues during the different stages of goat lactation., Results: A total of 1,938 (1,111 upregulated, 827 downregulated) differentially modified phosphorylation sites of 1,172 proteins were identified (P values < 0.05 and fold change of phosphorylation ratios > 1.5). Multiple phosphorylation sites of FASN, ACACA, mTOR, PRKAA, IRS1, RPS6KB, EIF4EBP1, JUN, and TSC2 were different in PL compared with LL. In addition, the Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis showed that the calcium signaling pathway, oxytocin signaling pathway and MAPK signaling pathway were enriched. The western blot results showed that the phosphorylation levels of ACACA (Ser80), EIF4EBP1 (Thr46) and IRS1 (Ser312) increased and JUN (Ser63) decreased in PL compared with LL. These results were consistent with the phosphoproteome results., Conclusions: In this study, we identified for the first time the differentially modified phosphorylation sites in goat mammary tissues between PL and LL. These results indicate that the multiple differentially modified phosphorylation sites of FASN, ACACA, mTOR, PRKAA, IRS1, RPS6KB, EIF4EBP1, TSC2, and JUN and proteins involved in the calcium signaling pathway, oxytocin signaling pathway, and MAPK signaling pathway are worthy of further exploration., (© 2021. The Author(s).)

Simple Summary: Dairy goats, as an important economic livestock for the milk supply in dairy products, have been raised worldwide. However, with the intensive management of dairy goat breeding, the disease problems are becoming increasingly serious, among which Corynebacterium pseudotuberculosis has been spreading in dairy goat breeding in China. In this study, we analyzed the transcriptome and proteomics of dairy goats infected with Corynebacterium pseudotuberculosis at 72 h and 144 h. Through a comparative correlation analysis of the data, we hope to find the pathogenic mechanism and provide a theoretical basis for the disease problems in dairy goat breeding. Corynebacterium pseudotuberculosis (C. pseudotuberculosis) is a zoonotic chronic infectious disease. It mainly occurs in dairy goats reared in herds, and once it invades the dairy goats, it is difficult to completely remove it, causing great harm to the development of the sheep industry. This study mainly was based on TMT-based quantitative proteomics and RNA-seq methods to measure the spleen samples of infected dairy goats at different time periods. Nine four-month-old dairy goats were divided into three groups, with three goats in each group. The dairy goats in the first group (NC group) were inoculated with 1.0 mL of sterilized normal saline subcutaneously, and the second (72 h group) and third groups (144 h group) were inoculated with 1.0 mL of 1 × 107 cfu/mL bacterial solution subcutaneously in the neck. Significant changes in the protein and mRNA expression were observed in different infection and control groups. In the 72 h group, 85 genes with differential genes and proteins were up-regulated and 91 genes were down-regulated in this study. In the 144 h group, 38 genes with differential genes and proteins were up-regulated and 51 genes were down-regulated. It was found that 21 differentially expressed genes and proteins were co-up-regulated in the two groups. There were 20 differentially expressed genes and proteins which were co-down-regulated in both groups. The 72 h group were mainly enriched in protein processing in the endoplasmic reticulum, lysosome, amino sugar and nucleotide sugar metabolism and the estrogen signaling pathway. In the 144 h group, they were protein processing in the endoplasmic reticulum pathway which was enriched by mRNA–proteins pairs co-upregulated by the five pairs. The combined transcriptomic and proteomic analyses were performed to provide insights into the effects of C. pseudotuberculosis through several regulatory features and pathways. We found that in the early stage of infection (72 h), the co-upregulated gene–protein pairs were enriched in multiple pathways, which jointly defended against a bacterial invasion. However, in the later stages of infection (144 h), when the disease stabilizes, a few co-upregulated gene–protein pairs played a role in protein processing in the endoplasmic reticulum pathway. In addition, the mRNA and protein expressions of dairy goats infected with the bacteria at different periods of time indicated the adaptability of dairy goats to the bacteria. At the same time, it guides us to carry out a corresponding treatment and feeding management for dairy goats according to different periods of time. [ABSTRACT FROM AUTHOR]

Pou2F3 (POU class 2 homeobox 3) is found to be ubiquitously expressed in multiple epidermal layer cells to mediating proliferation. Although some POU factors exert a crucial regulation in mammary epithelial cells (MECs), the biological function of Pou2F3 is unclear. In this study, we aimed to investigate the endogenous potential effects of Pou2F3 on the proliferation and the roles of PI3K/AKT/mTOR signaling pathway in MECs. We used small interfering RNA to silence Pou2F3 expression. The interfering efficiency of Pou2F3 was confirmed by using RT-qPCR and Western blot. The cell viability and proliferation were indicated by Cell Counting Kit-8 and EdU assays. Flow cytometry was performed to evaluate the cell apoptosis in MECs. These results demonstrated that Pou2F3 potently suppressed the proliferation and induced the apoptosis of MECs. Consistently, the primary protein expressions of PI3K/AKT/mTOR signaling pathway were examined by Western blot. Pou2F3 silencing significantly increased the phosphorylation of PI3K, AKT and mTOR expressions. Moreover, Pou2F3 silencing reduced the ratio of BCL-2/BAX protein expression. Our findings show that Pou2F3 silencing can induce the proliferation of MECs and decrease the cell apoptosis, which suggest that Pou2F3 may serve as a potential upstream regulator of PI3K/AKT/mTOR signaling pathway in MECs. [ABSTRACT FROM AUTHOR]

Simple Summary: Yield and quality of goat milk are important indexes for screening dairy goat breeds. Therefore, it is necessary for us to improve the yield and quality of goat milk. In this study, we demonstrated that circRNA8220/miR-8516/STC2 could promote the synthesis of β-casein and triglyceride through PI3K/AKT/mTOR pathway. In addition, we found that circRNA8220/miR-8516/STC2 also promote proliferation via Ras/MEK/ERK pathway in goat mammary epithelial cells (GMECs). These findings contribute to a better understanding of circRNA-controlled breast development and lactation mechanisms and provide new potential insights into the regulation of breast development and milk composition in dairy goats. Circular RNAs (circRNAs), which are considered a large class of endogenous noncoding RNAs, function as regulators in various biological procedures. In this study, the function and molecular mechanisms of circRNA8220 in goat mammary epithelial cells (GMECs) were explored. CircRNA8220 could spong miR-8516 and block the function of miR-8516 by binding to the target site of miR-8516 a negative feedback relationship existed between circRNA8220 and miR-8516. Stanniocalcin 2 (STC2) was a target gene of miR-8516. circRNA8220 could up-regulate the expression of STC2 by sponging miR-8516 in GMECs. circRNA8220/miR-8516/STC2 could promote proliferation and enhance the synthesis of β-casein and triglycerides (TG) via Ras/MEK/ERK and PI3K/AKT/mTOR signaling pathways, respectively. [ABSTRACT FROM AUTHOR]