UFL1 regulates milk protein and fat synthesis–related gene expression of bovine mammary epithelial cells probably via the mTOR signaling pathway

2021 ◽  
Author(s):  
Chengmin Li ◽  
Lian Li ◽  
Ilyas Ali ◽  
Meiqian Kuang ◽  
Xinling Wang ◽  
...  
Keyword(s):  
Gene Expression ◽  
Epithelial Cells ◽  
Signaling Pathway ◽  
Milk Protein ◽  
Mammary Epithelial Cells ◽  
Mammary Epithelial ◽  
Related Gene ◽  
Mtor Signaling Pathway ◽  
Bovine Mammary Epithelial Cells ◽  
Fat Synthesis

scholarly journals  Gene expression of six major milk proteins in primary bovine mammary epithelial cells isolated from milk during the first twenty weeks of lactation

2012 ◽  
Vol 57 (No. 10) ◽  
pp. 469-480 ◽  
Author(s):  
T. Sigl ◽  
H.H.D. Meyer ◽  
S. Wiedemann
Keyword(s):  
Gene Expression ◽  
Epithelial Cells ◽  
Milk Protein ◽  
Protein Gene ◽  
Mammary Epithelial Cells ◽  
Mammary Epithelial ◽  
Milk Protein Gene ◽  
Bovine Mammary Epithelial Cells ◽  
Milk Protein Genes ◽  
Milk Protein Gene Expression

&nbsp;The objective of the present study was to refine a previously developed method to isolate primary bovine mammary epithelial cells (pBMEC) from fresh milk. Using this method, it was tested whether the number of pBMEC and the relation of recovered pBMEC to total somatic cell count vary within the individual lactation stages. Furthermore, the expression levels of the milk protein genes during the first twenty weeks of lactation were determined by quantitative PCR method. A total number of 152 morning milk samples were obtained from twenty-four Holstein-Friesian cows during the first 20 weeks of lactation (day 8, 15, 26, 43, 57, 113, and 141 postpartum). Numbers of extracted pBMEC were consistent at all time-points (1.1 &plusmn; 0.06 to 1.4 &plusmn; 0.03 &times;10<sup>3</sup>/ml) and an average value of RNA integrity number (RIN) was 6.3 &plusmn; 0.3. Percentage of pBMEC in relation to total milk cells (2.0 &plusmn; 0.2 to 6.7 &plusmn; 1.0%) correlated with milk yield. Expression patterns of the casein genes alpha (&alpha;)<sub>S1</sub>, (&alpha;)<sub>S2</sub>, beta (&beta;), and kappa (&kappa;) (CSN1S1, CSN1S2, CSN2, CSN3, respectively) and the whey protein genes &alpha;-lactalbumin (LALBA) and progestagen-associated endometrial protein (PAEP; known as &beta;-lactoglobulin) were shown to be comparable, i.e. transcripts of all six milk protein genes were found to peak during the first two weeks of lactation and to decline continuously towards mid lactation. However, mRNA levels were different among genes with CSN3 showing the highest and LALBA the lowest abundance. We hypothesized that milk protein gene expression has a pivotal effect on milk protein composition with no influence on milk protein concentration. This paper is the first to describe milk protein gene expression during lactation in pBMEC collected in milk. Future studies will be needed to understand molecular mechanisms in pBMEC including regulation of expression and translation throughout lactation. &nbsp;

scholarly journals Bta-miR-34b controls milk fat biosynthesis via the Akt/mTOR signaling pathway by targeting RAI14 in bovine mammary epithelial cells

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Yujuan Wang ◽  
Xiaoyu Wang ◽  
Meng Wang ◽  
Li Zhang ◽  
Linsen Zan ◽  
...  
Keyword(s):  
Dairy Cows ◽  
Signaling Pathway ◽  
Milk Fat ◽  
Mammary Epithelial Cells ◽  
Mammary Epithelial ◽  
Mtor Signaling ◽  
Mtor Signaling Pathway ◽  
Bovine Mammary Epithelial Cells ◽  
Fat Synthesis ◽  
Milk Fat Synthesis

Abstract Background The biosynthesis of milk fat affects both the technological properties and organoleptic quality of milk and dairy products. MicroRNAs (miRNAs) are endogenous small non-coding RNAs that inhibit the expression of their mRNA targets and are involved in downstream signaling pathways that control several biological processes, including milk fat synthesis. miR-34b is a member of the miR-34 miRNA cluster, which is differentially expressed in the mammary gland tissue of dairy cows during lactation and dry periods. Previous studies have indicated miR-34b is a potential candidate gene that plays a decisive role in regulating milk fat synthesis; therefore, it is important to focus on miR-34b and investigate its regulatory effect on the biosynthesis of milk fat in bovine mammary epithelial cells (BMECs). Results In this study, elevated miR-34b levels reduced milk fat synthesis, upregulated 1,999 genes, and downregulated 2,009 genes in BMECs. Moreover, Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis of differentially expressed genes suggested that miR-34b may play an inhibitory role in milk fat synthesis via the protein kinase B (Akt)/mammalian target of rapamycin (mTOR) signaling pathway by reducing phosphorylation levels. Notably, the mTOR activator MHY1485 rescued the inhibitory effect of miR-34b. Furthermore, we demonstrated that retinoic acid-induced protein 14 (RAI14) is a target of miR-34b via TargetScan and immunofluorescence assays. RAI14 mRNA and protein levels were significantly decreased by the miR-34b mimic and increased by the miR-34b inhibitor. Moreover, the reduction in RAI14 levels led to the inhibition of the Akt/mTOR signaling pathway. Conclusions Overall, our results identified a miR-34b-RAI14-Akt/mTOR regulatory network, while also providing a theoretical basis for the molecular breeding of dairy cows.

Examination of methionine stimulation of gene expression in dairy cow mammary epithelial cells using RNA-sequencing

2020 ◽  
Vol 87 (2) ◽  
pp. 226-231
Author(s):  
Xiaoming Hou ◽  
Minghui Jiang ◽  
Jinyu Zhou ◽  
Shuyuan Song ◽  
Feng Zhao ◽  
...  
Keyword(s):  
Gene Expression ◽  
Protein Synthesis ◽  
Epithelial Cells ◽  
Rna Sequencing ◽  
Milk Protein ◽  
Mammary Epithelial Cells ◽  
Cell Model ◽  
Expression Profiles ◽  
Mammary Epithelial ◽  
Bovine Mammary Epithelial Cells

AbstractIn this research communication, a cell model with elevated β-CASEIN synthesis was established by stimulating bovine mammary epithelial cells with 0.6 mM methionine, and the genome-wide gene expression profiles of methionine-stimulated cells and untreated cells were investigated by RNA sequencing. A total of 458 differentially expressed genes (DEGs; 219 upregulated and 239 downregulated) were identified between the two groups. Gene Ontology (GO) analysis showed that the two highest-ranked GO terms in ‘molecular function’ category were ‘binding’ and ‘catalytic activity’, suggesting that milk protein synthesis in methionine-stimulated cells requires induction of gene expression to increase metabolic activity. Kyoto Encyclopedia of Genes and Genomes analysis revealed that within the ‘environmental information processing’ category, the subcategory that is most highly enriched for DEGs was ‘signal transduction’. cGMP-PKG, Rap1, calcium, cAMP, PI3K-AKT, MAPK, and JAK-STAT are the pathways with the highest number of DEGs, suggesting that these signaling pathways have potential roles in mediating methionine-induced milk protein synthesis in bovine mammary epithelial cells. This study provides valuable insights into the physiological and metabolic adaptations in cells stimulated with methionine. Understanding the regulation of this transition is essential for effective intervention in the lactation process.

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