Background: Interest is increasing in the role of peptide-bound amino acids (AAs) in milk protein synthesis because studies have found that the uptake of some essential AAs by the mammary gland cannot meet the requirements for milk protein synthesis. Although the role of dipeptide in milk protein synthesis is clearly established, little is known about the underlying mechanisms. Objective: The objective of this study was to determine whether small peptides can be taken up intact by the peptide transporters in mammary tissue explants and the underlying mechanisms of the effects of methionyl-methionine (MetMet) supplementation on milk protein synthesis. Methods: Mammary tissue explants were cultured in conditional medium and then treated with different concentrations of Met-Met that replace 0%, 5%, 10%, 15%, 20%, and 25% of free Met for another 24 h. In some experiments, explants were cultured with an optimal dose of Met-Met with or without the inhibitors of peptide transporter 2 [PepT2; diethylpyrocarbonate (DEPC), 0.1 mmol/L] and aminopeptidase N (APN; bestatin, 20 mmol/L) for 24 h. Results: The substitutions of 15% free Met with Met-Met significantly promoted a-s1 casein (as1-CN) expression in the mammary explants (P < 0.05). The inhibition of the PepT2 by DEPC or APN by bestatin significantly decreased the MetMet‐stimulated increase of as1-CN expression (P < 0.05). Compared with the control group (0% Met-Met), absorption of Val, Met, Leu, Phe, Lys, and His was improved, and mRNA abundance of the neutral and basic AA transporter was increased in the 15% Met-Met group (P < 0.05). In addition, the mRNA abundance of the mammalian target of rapamycin (mTOR), p70 ribosomal S6 kinase 1 gene, eukaryotic initiation factor 4E binding protein 1 gene , Janus kinase 2 (JAK2), and signal transducer and activator of transcription 5 (STAT5) was increased in the 15% Met-Met‐treated group (P < 0.05). Conclusion: Met-Met promoted as1-CN synthesis in cultured bovine mammary gland explants, and this stimulation may be mediated by enhanced intracellular substrate availability and by activating JAK2-STAT5 and mTOR signaling pathways. J Nutr doi: 10.3945/jn.114.208330.
Last. Jianxin Liu(ZJU: Zhejiang University)H-Index: 33
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Summary This study was conducted to investigate the effects of phenylalanine (Phe) and threonine (Thr) oligopeptides on αs1 casein gene expression and milk protein synthesis in bovine mammary epithelial cells. Primary mammary epithelial cells were obtained from Holstein dairy cows and incubated in Dulbecco's modified Eagle's medium-F12 medium (DMEM/F12) containing lactogenic hormones (prolactin and glucocorticoids). Free Phe (117 μg/ml) was substituted partly with peptide-bound Phe (phenylalanyl...
Last. Weihong Wang(SDU: Shandong University)H-Index: 1
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In this work, an electrophoretically mediated microanalysis (EMMA) method with a partial-filling technique was setup to evaluate the inhibitory potency of novel compounds toward aminopeptidase N (APN). It was necessary to optimize the electrophoretic conditions with respect to the kinetic constraints and for attaining high sensitivity. In our setup, a part of the capillary was filled with the incubation buffer for the enzyme reaction, whereas the rest was filled with a suitable BGE for the separ...
Last. David Smith(UM: University of Michigan)H-Index: 135
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This study evaluated the developmental gene and protein expression of proton-coupled oligopeptide transporters (POTs: PepT1, PepT2, PhT1 and PhT2) in different regions of rodent brain, and the age-dependent uptake of a POT substrate, glycylsarcosine, in brain slices. Slices were obtained from cerebral cortex, cerebellum and hypothalamus of wildtype and PepT2 null mice, and from rats at different ages. Gene and protein expression were determined by real-time PCR and immunoblot analyses. Brain sli...
Amino acid (AA) transporters may act as sensors, as well as carriers, of tissue nutrient supplies. This review considers recent advances in our understanding of the AA-sensing functions of AA transporters in both epithelial and nonepithelial cells. These transporters mediate AA exchanges between extracellular and intracellular fluid compartments, delivering substrates to intracellular AA sensors. AA transporters on endosomal (eg, lysosomal) membranes may themselves function as intracellular AA s...
Translation initiation is known to be regulated by the binding of eukaryotic initiation factor 4E (eIF4E) by binding proteins (4EBPs), and there is evidence that amino acid deprivation and other cellular stresses upregulate 4EBP1 expression. To pursue the question of whether diets limited in an essential amino acid lead to induction of 4EBP1 expression in vivo, diets that varied in methionine and cystine content were fed to rats for 7 days, and 4EBP1 mRNA and protein levels and 4EBP1 phosphoryla...
Last. Jianxin Liu(ZJU: Zhejiang University)H-Index: 33
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As the main precursor for lactose synthesis, large amounts of glucose are required by lactating dairy cows. Milk yield greatly depends on mammary lactose synthesis due to its osmoregulatory property for mammary uptake of water. Thus, glucose availability to the mammary gland could be a potential regulator of milk production. In the present study, the effect of glucose availability on expression of the key genes involved in synthesis of milk fat, lactose and glucose metabolism in vitro was invest...
The transcription factor STAT5A is essential for two processes during mammary gland development. First, it controls the development of luminal progenitor cells from stem cells1 and second, it has a role during pregnancy where it is required for alveologenesis2,3 the production of clusters of luminal cells that synthesize and secrete milk during lactation. Thus, deletion of STAT5A in late pregnancy results in lactation failure. Alveologenesis requires the proliferation of a different lineage of l...
Development of the mammary gland requires the coordinated action of proteolytic enzymes during two phases of remodelling. Firstly, new ducts and side-branches thereof need to be established during pregnancy to generate an extensive ductal tree allowing the secretion and transport of milk. A second wave of remodelling occurs during mammary involution after weaning. We have analysed the role of the cell surface protease aminopeptidase N (Anpep, APN, CD13) during these processes using Anpep deficie...
Last. Jianxin Liu(ZJU: Zhejiang University)H-Index: 33
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Summary This study was conducted to investigate the expression of oligopeptide transporter 2 (PepT2) and its potential function in bovine mammary gland. First, the PepT2 mRNA and protein were determined in cultured mammary epithelial cells. Then the effects of lactogenic hormones (prolactin, hydrocortisone or insulin) and substrate (threonyl-phenylalanyl-phenylalanine) on PepT2 were investigated. The PepT2 mRNA and protein were successfully detected in bovine mammary epithelial cells. PepT2 gene...
#1Massimo Bionaz(UIUC: University of Illinois at Urbana–Champaign)H-Index: 34
#2Juan J. Loor(UIUC: University of Illinois at Urbana–Champaign)H-Index: 68
A crucial role for both insulin and mTOR in the regulation of milk protein synthesis is emerging. Bovine mammary biopsies harvested during late-pregnancy through end of subsequent lactation were used to evaluate via quantitative PCR the expression of 44 genes involved in pathways of insulin, mTOR, AMPK, and Jak2-Stat5 signalling and also glucose and amino acid (AA) transporters. We observed an increased expression during lactation of ELF5, AA and glucose transporters, insulin signaling pathway c...
Abstract Bovine mammary epithelial cells (BMECs) are essential for lactation in the dairy cow mammary gland, and are often used as a cellular model to study changes in inflammatory responses and lactation functions with exogenous stimuli. Prolactin (PRL) promotes milk protein synthesis by continuously activating the Janus kinase 2 and signal transducer and activator of transcription 5 (JAK2-STAT5) pathway. Lipopolysaccharides (LPS) activates inflammatory responses in cells and inhibits casein sy...
Our previous transcriptomic study found that methionyl-methionine (Met-Met) exerts an anti-inflammatory effect in the bovine mammary epithelial cell (MAC-T) at a molecular level. However, evidence of whether the metabolic production of Met-Met confers protection was scarce. To investigate the inflammatory response and metabolite changes of Met-Met in lipopolysaccharide (LPS)-induced inflammation of MAC-T, mass spectrometry-based metabolomics and qPCR were conducted. The increased levels of IL-8,...
Crop milk is the sole source of nutrition that sustains young pigeons (squabs) throughout growth and development. Protein accounts for approximately 55% of the nutrients in crop milk; however, its regulation mechanism remains unclear. In our study, three experiments were conducted to investigate the possible underlying mechanism of crop milk protein synthesis and nutritional interventions. Isobaric tagging for relative and absolute quantification (iTRAQ) analysis found that the Janus activated k...
The aim of this study was to assess the effect of heat stress on dairy cow performance and on the expression of selected genes involved in milk protein metabolism. Eight Italian Holstein Friesian cows were kept under thermoneutral conditions (temperature-humidity index (THI) 0.05), CSN3 (p > 0.05), HSPA8 (p > 0.05), and STAT5B (p > 0.05) mRNA. Mild heat stress reduced the performance of dairy cows without affecting the expression of genes coding for caseins.
Milk production is economically important to the Brazilian agribusiness, and the majority of the country's milk production derives from Girolando (Gir × Holstein) cows. This study aimed to identify quantitative trait loci (QTL) and candidate genes associated with 305-d milk yield (305MY) in Girolando cattle. In addition, we investigated the SNP-specific variances for Holstein and Gir breeds of origin within the sequence of candidate genes. A single-step genomic BLUP procedure was used to identif...
#1Bingna Cai(CAS: Chinese Academy of Sciences)H-Index: 6
#2Peng Wan(CAS: Chinese Academy of Sciences)H-Index: 6
Last. Jianyu Pan(CAS: Chinese Academy of Sciences)H-Index: 6
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Octopus protein hydrolysate has been reported to increase milk yield and milk protein production. In this paper, the utilization and underlying mechanisms of bioactive peptide fractions from octopus protein hydrolysate on β-casein expression in mouse mammary epithelial cells (HC11) were investigated. Fraction OPH3-1 significantly stimulated cell proliferation and β-casein synthesis in HC11 cells, which was purified by ultra-filtration and gel-filtration chromatography. The MWs of the peptides fr...
Last. Hongyun Liu(ZJU: Zhejiang University)H-Index: 14
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Bovine peptide transporter 2 (bPepT2), which mediates the absorption of di tripeptides in the bovine mammary gland, was predicted to contain multiple putative N-glycosylation sites of asparagine residues. N-Linked glycosylation is proven to be essential for the folding, stability, localization, and substrate binding of nutrient transporters and could therefore potentially have an essential role in the function of bPepT2. This study investigated the effect of mutagenesis of N-glycosylation sites ...
AMP-activated protein kinase (AMPK) senses the cellular energy status and coordinates catabolic and anabolic processes. Extensive studies have proposed that AMPK regulates energy homeostasis, cell growth, autophagy, mitochondrial biology and inflammation. The biological functions of AMPK vary in different tissues or organs. As a unique organ that produces milk, the mammary gland has recently attracted substantial research attention. This review discusses how AMPK in the mammary gland is activate...
Last. Hongyun Liu(ZJU: Zhejiang University)H-Index: 14
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BACKGROUND: Pregnancy-induced hypoaminoacidemia, l-methionine (Met) included, disturbs embryogenesis and may also affect breast function. Supplementation with the dipeptide l-methionyl-Met (Met-Met) may improve lactation performance. OBJECTIVE: We compared the effects of supplemental Met or Met-Met during pregnancy on mammogenesis and lactogenesis and investigated underlying mechanisms. METHODS: In experiment 1, 9-wk-old ICR mice (n = 72, approximately 30 g) were divided into 3 groups. During th...
Ketosis is a metabolic disease of dairy cows often characterized by high concentrations of ketone bodies and fatty acids, but low milk protein and milk production. The Janus kinase 2 (JAK2)-signal transducer and activator of transcription 5 (STAT5) and the mechanistic target of rapamycin (mTOR) signaling pathways are central for the regulation of milk protein synthesis. The effect of high levels of fatty acids on these pathways and beta-casein synthesis are unknown in dairy cows with clinical ke...