scholarly journals A Functional 3′ UTR Polymorphism of FADS2 Affects Cow Milk Composition through Modifying Mir-744 Binding

Animals ◽  
2019 ◽  
Vol 9 (12) ◽  
pp. 1090 ◽  
Author(s):  
Mingxun Li ◽  
Xubin Lu ◽  
Qisong Gao ◽  
Mengqi Wang ◽  
Abdelaziz Adam Idriss Arbab ◽  
...  
Keyword(s):  
Mammary Epithelial Cells ◽  
Specific Inhibitor ◽  
Milk Composition ◽  
Mammary Epithelial ◽  
Cow Milk ◽  
Bovine Mammary Epithelial Cells ◽  
Protein Levels ◽  
Omega 6 ◽  
Delta 6 Desaturase ◽  
Consequent Increase

This study determined the associations of FADS2 c.1571G>A with milk FAs content and revealed that cows with the GG genotype had improved levels of delta-6 desaturase substrates (linoleic acid, C18:2n-6; p < 0.001) and decreased levels of desaturase products (gamma-linolenic acid, C18:3n-6; p < 0.001), indicating a reduction in FADS2 expression or delta-6 desaturase activity caused by this polymorphism. Computer alignment demonstrated that c.1571G>A occurred within a potential miR-744 binding site. When the c.1571G allele was present, the luciferase activity of reporter constructs was significantly suppressed by miR-744, while no such effect was observed with the A allele. Overexpression of miR-744 in bovine mammary epithelial cells (with the 1571GG genotype) downregulated FADS2 expression at both mRNA and protein levels. In contrast, inhibition of endogenous miR-744 with a specific inhibitor dramatically upregulated FADS2 expression. Taken together, these lines of evidence indicated that the c.1571A minor allele abolished the ability of miR-744 to bind FADS2, with a consequent increase in FADS2 expression levels and synthesis of omega-6 LC-PUFAs.

Mammary gland copper transport is stimulated by prolactin through alterations in Ctr1 and Atp7A localization

2006 ◽  
Vol 291 (4) ◽  
pp. R1181-R1191 ◽  
Author(s):  
Shannon L. Kelleher ◽  
Bo Lönnerdal
Keyword(s):  
Plasma Membrane ◽  
Mammary Gland ◽  
Molecular Mechanisms ◽  
Mammary Epithelial Cells ◽  
Milk Composition ◽  
Mammary Epithelial ◽  
Recycling Endosome ◽  
Protein Levels ◽  
Stage Of Lactation ◽  
Golgi Network

Milk copper (Cu) concentration declines and directly reflects the stage of lactation. Three Cu-specific transporters (Ctr1, Atp7A, Atp7B) have been identified in the mammary gland; however, the integrated role they play in milk Cu secretion is not understood. Whereas the regulation of milk composition by the lactogenic hormone prolactin (PRL) has been documented, the specific contribution of PRL to this process is largely unknown. Using the lactating rat as a model, we determined that the normal decline in milk Cu concentration parallels declining Cu availability to the mammary gland and is associated with decreased Atp7B protein levels. Mammary gland Cu transport was highest during early lactation and was stimulated by suckling and hyperprolactinemia, which was associated with Ctr1 and Atp7A localization at the plasma membrane. Using cultured mammary epithelial cells (HC11), we demonstrated that Ctr1 stains in association with intracellular vesicles that partially colocalize with transferrin receptor (recycling endosome marker). Atp7A was primarily colocalized with mannose 6-phosphate receptor (M6PR; late endosome marker), whereas Atp7B was partially colocalized with protein disulfide isomerase (endoplasmic reticulum marker), TGN38 ( trans-Golgi network marker) and M6PR. Prolactin stimulated Cu transport as a result of increased Ctr1 and Atp7A abundance at the plasma membrane. Although the molecular mechanisms responsible for these posttranslational changes are not understood, transient changes in prolactin signaling play a role in the regulation of mammary gland Cu secretion during lactation.

14-3-3γaffects eIF5 to regulateβ-casein synthesis in bovine mammary epithelial cells

2016 ◽  
Vol 96 (4) ◽  
pp. 478-487
Author(s):  
Cuiping Yu ◽  
Chaochao Luo ◽  
Xinyu Gu ◽  
Yanli Zang ◽  
Bo Qu ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Regulatory Mechanism ◽  
Mammary Epithelial Cells ◽  
Mammary Epithelial ◽  
Initiation Factor ◽  
Biological Processes ◽  
Bovine Mammary Epithelial Cells ◽  
Protein Levels ◽  
Casein Synthesis

The 14-3-3γ protein participates in many biological processes; however, its regulatory mechanism in milk protein synthesis is not well studied. We hypothesized that 14-3-3γ might affect eIF5 (an initiation factor) to regulate β-casein synthesis in dairy cows. In this study, a possible interaction between 14-3-3γ and eIF5 was investigated using bovine mammary epithelial cells (BMECs). The expression levels of 14-3-3γ and eIF5 in the mammary gland tissues from cows producing higher quality milk were higher than those from cows producing low-quality milk. Moreover, the expression of 14-3-3γ, eIF5, and β-casein were increased at both mRNA and protein levels in BMECs cultured in vitro with methionine (Met) supplementation. Coimmunoprecipitation, colocalization, and FRET analysis further showed the evidences that 14-3-3γ physically bound to eIF5 in BMECs. Gene function studies revealed that 14-3-3γ positively regulated eIF5 through alteration of eIF2α/p-eIF2α ratio. Collectively, our data suggest that 14-3-3γ regulates β-casein translation in BMECs through interaction with eIF5.

scholarly journals Establishment of inflammatory model of bovine mammary epithelial cells induced by lipoteichoic acid

2020 ◽  
pp. 31-37
Author(s):  
Xu Ping ◽  
Tetiana Fotina ◽  
Hanna Fotina ◽  
Sanhu Wang
Keyword(s):  
Epithelial Cells ◽  
Pathogenic Bacteria ◽  
Mammary Epithelial Cells ◽  
Lipoteichoic Acid ◽  
Mammary Epithelial ◽  
Cow Milk ◽  
Cytotoxic Factor ◽  
Bovine Mammary Epithelial Cells ◽  
Tnf Α ◽  
Wide Range

The mammary gland of the cow is particularly susceptible to infections of a wide range of pathogenic bacteria, including both Gram-positive and Gram-negative bacteria. The endotoxins of these pathogenic bacteria include peptidoglycan (PGN), lipoteichoic acid (LTA) and lipopolysaccharide (LPS), and they are the pathogen-associated molecular patterns (PAMPs) to induce mastitis. Cow mastitis is a detrimental factor in dairy farming industry. Lipoteichoic acid (LTA) is the main component of Staphylococcus aureus cell wall and the key cytotoxic factor causing inflammation. The aims of our work was to establish inflammatory model of study procedures were approved by the Animal Care and Use Committee of the Sumy National Agricultural University, Sumy, Ukraine, and the Henan Institute of Science and Technology, Xinxiang, China, and performed in accordance with the animal welfare and ethics guidelines. The BMECs harvested from mid-lactation dairy cow milk were isolated by our laboratory. Briefly, the base medium for this cell is DMEM/F-12 (Gibco, USA, cat.12400-024). The complete growth medium included 10% fetal bovine serum (Biological Industries, Israel, cat.04-011-1A/B), DMEM/F-12, and 10 ng/mL epidermal growth factor (Sigma, USA, cat. E4127). Cells were maintained at 37℃in an incubator containing 5% CO2. When cells grew to 80% confluency, the cells were rinsed twice with PBS, and then the primary mammary epithelial cells were trypsinized with 0.25% trypsin plus 0.02% EDTA and passaged. In this study, one inflammatory bovine mammary epithelial cell (BMEC) model was established by infecting the cells with LTA. The BMEC viability induced by LTA were evaluated. The expressions of pro-inflammatory cytokines (TNF-α and IL-6) were measured by ELISA and RT- qPCR. The results showed that the treatment of BMECs with LTA at 20 ng/μL for 24 h obviously improved TNF-α and IL-6 protein and gene expression levels. The establishment of the model will play an important role in the screening of anti-inflammatory drugs and the study of the mechanism of action in the future.

scholarly journals 5-ALA Attenuates the Palmitic Acid-Induced ER Stress and Apoptosis in Bovine Mammary Epithelial Cells

Molecules ◽  
2021 ◽  
Vol 26 (4) ◽  
pp. 1183
Author(s):  
Mst Mamuna Sharmin ◽  
Md Aminul Islam ◽  
Itsuki Yamamoto ◽  
Shin Taniguchi ◽  
Shinichi Yonekura
Keyword(s):  
T Cells ◽  
Epithelial Cells ◽  
Palmitic Acid ◽  
Er Stress ◽  
Aminolevulinic Acid ◽  
Mammary Epithelial Cells ◽  
Mammary Epithelial ◽  
Protective Effects ◽  
Bovine Mammary Epithelial Cells ◽  
Induced Apoptosis

The conservation of mammary gland physiology by maintaining the maximum number of mammary epithelial cells (MECs) is of the utmost importance for the optimum amount of milk production. In a state of negative energy balance, palmitic acid (PA) reduces the number of bovine MECs. However, there is no effective strategy against PA-induced apoptosis of MECs. In the present study, 5-aminolevulinic acid (5-ALA) was established as a remedial agent against PA-induced apoptosis of MAC-T cells (an established line of bovine MECs). In PA-treated cells, the apoptosis-related genes BCL2 and BAX were down- and upregulated, respectively. The elevated expression of major genes of the unfolded protein response (UPR), such as CHOP, a proapoptotic marker (C/EBP homologous protein), reduced the viability of PA-treated MAC-T cells. In contrast, 5-ALA pretreatment increased and decreased BCL2 and BAX expression, respectively. Moreover, cleaved caspase-3 protein expression was significantly reduced in the 5-ALA-pretreated group in comparison with the PA group. The downregulation of major UPR-related genes, including CHOP, extended the viability of MAC-T cells pretreated with 5-ALA and also reduced the enhanced intensity of the PA-induced expression of phospho-protein kinase R-like ER kinase. Moreover, the enhanced expression of HO-1 (antioxidant gene heme oxygenase) by 5-ALA reduced PA-induced oxidative stress (OxS). HO-1 is not only protective against OxS but also effective against ER stress. Collectively, these findings offer new insights into the protective effects of 5-ALA against PA-induced apoptosis of bovine MECs.

Exogenous phospholipase A2 affects inflammatory gene expression in primary bovine mammary epithelial cells

2019 ◽  
Vol 86 (2) ◽  
pp. 177-180
Author(s):  
Jacqueline P. Kurz ◽  
Mark P. Richards ◽  
Matthew Garcia ◽  
Zhongde Wang
Keyword(s):  
Epithelial Cells ◽  
Phospholipase A2 ◽  
Inflammatory Responses ◽  
Mammary Epithelial Cells ◽  
Constitutive Expression ◽  
Mammary Epithelial ◽  
Inflammatory Factors ◽  
Bovine Mammary Epithelial Cells ◽  
Lps Challenge

AbstractThis Research Communication addresses the hypothesis that exogenously administered phospholipase A2 (PLA2) affects the inflammatory responses of bovine mammary epithelial cells (bMEC) in vitro with the aim of providing preliminary justification of investigation into the uses of exogenously administered PLA2 to manage or treat bovine mastitis. Primary bMEC lines from 11 lactating Holstein dairy cows were established and the expression of 14 pro-inflammatory genes compared under unchallenged and lipopolysaccharide (LPS)-challenged conditions, with and without concurrent treatment with bovine pancreatic PLA2G1B, a secreted form of PLA2. No differences in the expression of these genes were noted between PLA2-treated and untreated bMEC under unchallenged conditions. Following LPS challenge, untreated bMEC exhibited significant downregulation of CXCL8, IL1B, CCL20, and CXCL1. In contrast, PLA2-treated bMEC exhibited significant downregulation of IL1B and CCL20 only. These findings indicate that exogenous PLA2 affects the expression of some pro-inflammatory factors in immune-stimulated bMEC, but does not influence the constitutive expression of these factors. Further investigation of the influence of exogenous PLA2 in the bovine mammary gland is justified.

Cell cycle regulation of mammary epithelial cell detachment byStaphylococcus aureus

1996 ◽  
Vol 63 (4) ◽  
pp. 543-553 ◽  
Author(s):  
Boris Zavizion ◽  
Andrew J. Bramley ◽  
Ioannis Politis
Keyword(s):  
Cell Cycle ◽  
Epithelial Cells ◽  
Culture Medium ◽  
Mammary Epithelial Cells ◽  
Mammary Epithelial ◽  
Cell Detachment ◽  
Staph Aureus ◽  
Bovine Mammary Epithelial Cells ◽  
Isogenic Mutants ◽  
Cycle Regulation

SummaryThe effect ofStaphylococcus aureuson detachment of bovine mammary epithelial cells in culture was examined. Mammary epithelial cells became detached from fresh monolayers following a 3 h incubation in the presence ofStaph. aureusM60. Two different procedures indicated that cell detachment coincided with the S-phase of the cell cycle. The roles of proteinases, toxins and Ca availability in inducing cell detachment were examined. Addition of the proteinase inhibitor phenyl-methylsulphonyl fluoride (1 mM) to the culture medium prevented cell detachment. Addition of a combination of purified staphylococcal proteinases XVI and XVII-B to the culture medium of mammary epithelial cells induced cell detachment in the absence ofStaph. aureus. Cell detachment may be caused by a staphylococcal proteinase. However, addition of Ca (10 mM) to the culture medium abolishedStaph. aureus-induced cell detachment, despite the fact that proteinase activity was still apparently present. Isogenic mutants ofStaph. aureusM60, expressing either ± or β toxins but not both, induced cell detachment, but to a lesser extent than the wild type. Thus, Ca and toxins play some role during cell detachment. Clones established from detached cells that were washed and replated showed the same susceptibility toStaph. aureus-induced cell detachment as the parental cells. This indicated that there is no subclone of mammary epithelial cells more sensitive to this effect.

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