Project description:Negative energy balance (NEB) is an altered metabolic state in high yielding cows that occurs during the first few weeks postpartum when energy demands for lactation and maintenance exceed the energy supply from dietary intake. NEB can, in turn, lead to metabolic disorders and to reduced fertility. Alterations in the expression of more than 700 hepatic genes have previously been reported in a study of NEB in postpartum dairy cows. miRNAs (microRNA) are known to mediate many alterations in gene expression post transcriptionally. To study the hepatic miRNA content of postpartum dairy cows, including their overall abundance and differential expression, in mild NEB (MNEB) and severe NEB (SNEB) short read RNA sequencing was carried out. A NEB dairy cow model developed previously was used. In this model differential feeding and milking regimes were used to produce two groups of Holstein Friesian cows; MNEB and SNEB. Briefly, MNEB cows were fed ad libitum grass silage with 8 kg/day of a 21% crude protein dairy concentrate and milked once daily. SNEB cows were fed 25 kg/day silage with 4 kg/day concentrate and milked thrice daily. All procedures were carried out under license in accordance with the European Community Directive, 86-609-EC. Cows were slaughtered approximately 14 days postpartum (MNEB; 13.6 ± 0.75, range 11–15; SNEB 14.3 ± 0.56, range 13–16 ) and the entire liver was removed within 15 to 30 min. Samples weighing approximately 1 g were dissected, rinsed in RNase-free phosphate buffer, snap-frozen in liquid nitrogen and stored at -80ºC. Liver tissue samples from 5 SNEB and 3 MNEB animals were used for miRNA library preparation.
Project description:We investigated miRNA expression in Holstein dairy cow of mammary gland with different producing quality milk using high-throughput sequence and qRT-PCR techniques. miRNA libraries were constructed from mammary gland tissues taken from a high producing quality milk and a low producing quality milk Holstein dairy cow, the small RNA digitalization analysis based on HiSeq high-throughput sequencing takes the SBS-sequencing by synthesis.The libraries included 4732 miRNAs. A total of 124 miRNAs in the high producing quality milk mammary gland showed significant differences in expression compared to low producing quality milk mammary gland (P<0.05). Conclusion: Our study provides a broad view of the bovine mammary gland small RNA expression profile characteristics. Differences in types and expression levels of miRNAs were observed between high producing quality milk and a low producing quality milk Holstein dairy cow
Project description:Negative energy balance (NEB) is an altered metabolic state in high yielding cows that occurs during the first few weeks postpartum when energy demands for lactation and maintenance exceed the energy supply from dietary intake. NEB can, in turn, lead to metabolic disorders and to reduced fertility. Alterations in the expression of more than 700 hepatic genes have previously been reported in a study of NEB in postpartum dairy cows. miRNAs (microRNA) are known to mediate many alterations in gene expression post transcriptionally. To study the hepatic miRNA content of postpartum dairy cows, including their overall abundance and differential expression, in mild NEB (MNEB) and severe NEB (SNEB) short read RNA sequencing was carried out.
Project description:Lipopolysaccharide (LPS) is a component of the outer membrane of gram-negative bacteria that is involved in pathogenic processes leading to postpartum metritis and endometritis in dairy cow. It also causes inflammation of the endometrium. This study aimed to investigate the effects of LPS on cow endometrial epithelial cells (BEECs). In this study, the label-free LC–MS/MS approach was employed to investigate the proteome profile of cow endometrial epithelial cells. A total of 1121 proteins were identified in the control group and model group, of which 186 differential proteins were accurately quantified. The proteomic results were verified by WB and IF techniques, and the same conclusion was obtained
Project description:Increased energy demands to support lactation, coupled with lowered feed intake capacity results in negative energy balance (NEB) and is typically characterized by extensive mobilization of body energy reserves in the early postpartum dairy cow. The catabolism of stored lipid leads to an increase in the systemic concentrations of nonesterified fatty acids (NEFA) and -hydroxy butyrate (BHB). Oxidation of NEFA in the liver result in the increased production of reactive oxygen species and the onset of oxidative stress and can lead to disruption of normal metabolism and physiology. The immune system is depressed in the peripartum period and early lactation and dairy cows are therefore more vulnerable to bacterial infections causing mastitis and or endometritis at this time. A bovine Affymetrix oligonucleotide array was used to determine global gene expression in the spleen of dairy cows in the early postpartum period. Spleen tissue was removed post mortem from five severe NEB (SNEB) and five medium NEB (MNEB) cows 15 days postpartum.SNEB increased systemic concentrations of NEFA and BHB, and white blood cell and lymphocyte numbers were decreased in SNEB animals. A total of 545 genes were altered by SNEB. Network analysis using Ingenuity Pathway Analysis revealed that SNEB was associated with NRF2-mediated oxidative stress, mitochondrial dysfunction, endoplasmic reticulum stress, natural killer cell signaling, p53 signaling, downregulation of IL-15, BCL-2, and IFN- ; upregulation of BAX and CHOP and increased apoptosis with a potential negative impact on innate and adaptive immunity.
Project description:This study was performed to discover the association between postpartum during negative energy balance and the expression of extracellular vesicle-coupled microRNAs signatures in follicular fluid of large follicle in dairy cows. For this, next-generation sequencing to total RNA miRNA was performed.
Project description:To explore potential functional genes related to milk production traits in cow, we performed total RNA-seq using liver biospy collected from 77 Holstein Friesian cows at postpartum 2 weeks.
Project description:This study aimed to investigate the effect of a mild underfeeding on global gene expression in the oviduct, the endometrium and the corpus luteum between 4 and 15 days post-ovulation in postpartum (PP) Holstein cows. Four control cows (C) received 100% of energy and protein requirements after calving when 4 underfed (U) received 80% of the control diet. Oestrus synchronization treatment was applied to induce ovulation on D80 PP. Oviduct, ovaries and the anterior part of each uterine horn were recovered chirurgically 4, 8, 12 and 15 days after ovulation. Gene expression was assessed on a dedicated 10K array and statistically analyzed on pools across the oestrus cycle days. Tissues appeared divergently "diet-responsive" as no differentially expressed gene (DEG) in the corpus luteum between U and C were highlighted, conversely to 293 DEGs in the oviduct vs. 1 in endometrium under a False discovery rate (FDR)<0.11, and 3830 DEGs vs. 28 respectively under a FDR<0.30. Pathways involved immunity, lipid catabolism and cell proliferation. However, once correlated with dedicated statistics (regularized canonical correlation analysis) to the evolution of 6 plasmatic markers measured weekly over 80 days, IGF-1, insulin, beta-Hydroxybutyrate and genes potentially implied in tissue repair mechanisms (e.g. FABP5/CRABP2, ELF1, TCEA1, NF-kB, TGF-beta1, HNF4A) became relevant to dairy cow physiology during PP in the oviduct and the endometrium. Keywords: Postpartum dairy cows, induced cycle, mild underfeeding, genital tract, gene-metabolites correlations
Project description:Milk production during the early postpartum in dairy cows can impact the circulating metabolites and negatively affects embryo survival. However, the molecular consequences for the embryo during its development in the oviduct are still unknown. The objective was to determine the impact of metabolic status on embryonic genome activation (EGA) using high-throughput sequencing to generate comprehensive transcriptome profiles of bovine 16-cell stage embryos which had undergone EGA in vitro or in the oviducts of primiparous postpartum dry or postpartum lactating Holstein Friesian cows.