Methionine (Met) can activate the mechanistic target of rapamycin (mTOR) to promote milk synthesis in mammary epithelial cells. However, it is largely unknown which G protein-coupled receptor can mediate the stimulation of Met on mTOR activation. In this study, we employed transcriptome sequencing to analyse which G protein-coupled receptors were associated with the role of Met and further used gene function study approaches to explore the role of G protein-coupled receptor 183 (GPR183) in Met stimulation on mTOR activation in HC11 cells. We identified nine G protein-coupled receptors including GPR183 whose expression levels were upregulated by Met treatment through RNA sequencing and subsequent quantitative real-time PCR analysis. Using GPR183 knockdown and overexpression technology, we demonstrate that GPR183 is a positive regulator of milk protein and fat synthesis and proliferation of HC11 cells. Met affected GPR183 expression in a dose-dependent manner, and GPR183 mediated the stimulation of Met (0·6 mM) on milk protein and fat synthesis, cell proliferation and mTOR phosphorylation and mRNA expression. The inhibition of phosphoinositide 3-kinase blocked the phosphorylation of mTOR and AKT stimulated by GPR183 activation. In summary, through RNA sequencing and gene function study, we uncover that GPR183 is a key mediator for Met to activate the phosphoinositide 3-kinase-mTOR signalling and milk synthesis in mouse mammary epithelial cells.

The G protein-coupled receptors (GPCR) sensing nutritional signals (amino acids, fatty acids, glucose, etc.) are not fully understood. In this research, we used transcriptome sequencing to analyse differentially expressed genes (DEG) in mouse mammary gland tissues at puberty, lactation and involution stages, in which eight GPCR were selected out and verified by qRT-PCR assay. It was further identified the role of GPR110-mediating nutrients including palmitic acid (PA) and methionine (Met) to improve milk synthesis using mouse mammary epithelial cell line HC11. PA but not Met affected GPR110 expression in a dose-dependent manner. GPR110 knockdown decreased milk protein and fat synthesis and cell proliferation and blocked the stimulation of PA on mechanistic target of rapamycin (mTOR) phosphorylation and sterol-regulatory element binding protein 1c (SREBP-1c) expression. In summary, these experimental results disclose DEG related to lactation and reveal that GPR110 mediates PA to activate the mTOR and SREBP-1c pathways to promote milk protein and fat synthesis.

Several amino acids can stimulate milk synthesis in mammary epithelial cells (MEC); however, the regulatory role of isoleucine (Ile) and underlying molecular mechanism remain poorly understood. In this study, we aimed to evaluate the regulatory effects of Ile on milk protein and fat synthesis in MEC and reveal the mediation mechanism of Brahma-related gene 1 (BRG1) on this regulation. Ile dose dependently affected milk protein and fat synthesis, mechanistic target of rapamycin (mTOR) phosphorylation, sterol regulatory element binding protein 1c (SREBP-1c) expression and maturation, and BRG1 protein expression in bovine MEC. Phosphatidylinositol 3 kinase (PI3K) inhibition by LY294002 treatment blocked the stimulation of Ile on BRG1 expression. BRG1 knockdown and gene activation experiments showed that it mediated the stimulation of Ile on milk protein and fat synthesis, mTOR phosphorylation, and SREBP-1c expression and maturation in MEC. ChIP-PCR analysis detected that BRG1 bound to the promoters of mTOR and SREBP-1c, and ChIP-qPCR further detected that these bindings were increased by Ile stimulation. In addition, BRG1 positively regulated the binding of H3K27ac to these two promoters, while it negatively affected the binding of H3K27me3 to these promoters. BRG1 knockdown blocked the stimulation of Ile on these two gene expressions. The expression of BRG1 was higher in mouse mammary gland in the lactating period, compared with that in the puberty or dry period. Taken together, these experimental data reveal that Ile stimulates milk protein and fat synthesis in MEC via the PI3K-BRG1-mTOR/SREBP-1c pathway.

Taurine (Tau) has many profound physiological functions, but its role and molecular mechanism in muscle cells are still not fully understood. In this study, we investigated the role and underlying molecular mechanism of Tau on protein synthesis and proliferation of C2C12 myoblast cells. Cells were treated with Tau (0, 60, 120, 180 and 240 μM) for 24 h. Tau dose-dependently promoted protein synthesis, cell proliferation, mechanistic target of rapamycin protein (mTOR) phosphorylation and also AT-rich interaction domain 4B (ARID4B) expression, with the best stimulatory effects at 120 μM. LY 294002 treatment showed that Tau promoted ARID4B expression in a phosphoinositide 3-kinase (PI3K)-dependent manner. ARID4B knockdown (by small interfering RNA transfection for 24 h) prevented Tau from stimulating protein synthesis and cell proliferation, whereas ARID4B gene activation (using the CRISPR/dCas9 technology) had stimulatory effects. ARID4B knockdown abolished Tau signalling to mRNA expression and protein phosphorylation of mTOR, whereas ARID4B gene activation had stimulatory effects. Chromatin immunoprecipitation (ChIP)-PCR identified that all of ARID4B, H3K27ac and H3K27me3 bound to the −4368 to –4591 bp site in the mTOR promoter, and ChIP-quantitative PCR (qPCR) further detected that Tau stimulated ARID4B binding to this site. ARID4B knockdown or gene activation did not affect H3K27me3 binding to the mTOR promoter but decreased or increased H3K27ac binding, respectively. Furthermore, ARID4B knockdown abolished the stimulation of Tau on H3K27ac binding to the mTOR promoter. In summary, these data uncover that Tau promotes protein synthesis and proliferation of C2C12 myoblast cells through the PI3K-ARID4B-mTOR pathway, providing a deep understanding of how Tau regulates anabolism in muscle cells.

Nanoscale magnetization modulation by electric field enables the construction of low-power spintronic devices for information storage applications and, etc. Electric field-induced ion migration can introduce desired changes in the material's stoichiometry, defect profile, and lattice structure, which in turn provides a versatile and convenient means to modify the materials’ chemical-physical properties at the nanoscale and in situ. In this review, we provide a brief overview on the recent study on nanoscale magnetization modulation driven by electric field-induced migration of ionic species either within the switching material or from external sources. The formation of magnetic conductive filaments that exhibit magnetoresistance behaviors in resistive switching memory via foreign metal ion migration and redox activities is also discussed. Combining the magnetoresistance and quantized conductance switching of the magnetic nanopoint contact structure may provide a future high-performance device for non-von Neumann computing architectures.

Phenotypic plasticity and rapid evolution are two important strategies by which invasive species adapt to a wide range of environments and consequently are closely associated with plant invasion. To test their importance in invasion success of Crofton weed, we examined the phenotypic response and genetic variation of the weed by conducting a field investigation, common garden experiments, and intersimple sequence repeat (ISSR) marker analysis on 16 populations in China. Molecular markers revealed low genetic variation among and within the sampled populations. There were significant differences in leaf area (LA), specific leaf area (SLA), and seed number (SN) among field populations, and plasticity index (PIv) for LA, SLA, and SN were 0.62, 0.46 and 0.85, respectively. Regression analyses revealed a significant quadratic effect of latitude of population origin on LA, SLA, and SN based on field data but not on traits in the common garden experiments (greenhouse and open air). Plants from different populations showed similar reaction norms across the two common gardens for functional traits. LA, SLA, aboveground biomass, plant height at harvest, first flowering day, and life span were higher in the greenhouse than in the open-air garden, whereas SN was lower. Growth conditions (greenhouse vs. open air) and the interactions between growth condition and population origin significantly affect plant traits. The combined evidence suggests high phenotypic plasticity but low genetically based variation for functional traits of Crofton weed in the invaded range. Therefore, we suggest that phenotypic plasticity is the primary strategy for Crofton weed as an aggressive invader that can adapt to diverse environments in China.

Leptin is an autocrine and paracrine factor which affects the development and function of mammary gland. The purpose of this study was to investigate the presence and regulatory effect of leptin in Chinese Guan Zhong dairy goat mammary gland from the virgin state to involution. The protein expression and localization of leptin and its long form receptor (OB-Rb) were detected by western blot and a confocal laser scanning microscope. Explants were cultured to detect the impacts of leptin on mammary gland, western blot was used to research leptin signal transduction pathway in pregnancy, lactation and involution. Leptin and amounts of OB-Rb protein were high throughout the virgin stage and at the beginning of pregnancy, lower at mid-pregnancy and throughout lactation, then higher at involution. Immunofluorescence performed with the anti-leptin and anti-leptin receptor antibody showed labelling located in adipose, epithelial cells, or extracellular matrix at different stages. The localization of leptin and OB-Rb revealed that leptin induced the expression of OB-Rb specifically and controlled the development and physiological function of the mammary gland by binding to OB-Rb. Leptin stimulated the proliferation and differentiation of ductal epithelial cells in pregnancy by JAK-MAPK signal pathway, enhanced the amount of β-casein in cultured lactating mammary gland by JAK-STAT5 signal pathway, made the mammary duct disappear and induced apoptosis of mammary epithelial cells and mammary gland restitution by JAK-STAT3 signal pathway in involution. Overall, this study demonstrated the importance and complexity of leptin and OB-Rb during mammary gland development and provides a valuable resource for future research in this area.