Project description:Effect of breed in mid lactation Holstein (H) and Montbéliarde (M) cows on mammary glande miRNA profile. Genetic polymorphisms are known to influence milk production and composition. However, genomic mechanisms involved in the genetic regulation of milk component synthesis are not completely understood. MicroRNAs (miRNA) regulate gene expression. The objective of the present study was to compare mammary gland miRNomes of two dairy cow breeds, Holstein and Montbéliarde, with different dairy performances. Milk, fat, protein, and lactose yields were lower in Montbéliarde than in Holstein cows. MiRNomes obtained using RNA-Seq technology from the mammary glands of Holstein (n = 5) and Montbéliarde (n = 6) lactating cows revealed 623 distinct expressed miRNAs, among which 596 were known and 27 were predicted miRNAs. The comparison of their abundance in the mammary gland of Holstein versus Montbéliarde cows showed 22 differentially expressed miRNAs (Padj ≤ 0.05). Among them, 11 presented a fold change ≥2, with 2 highly expressed miRNAs (miR-100 and miR-146b). Without taking into account the fold change, the differential miRNA with the highest abundance was miR-186, which is known to inhibit cell proliferation and epithelial-to-mesenchymal transition. Data mining showed that the 17 differentially expressed miRNAs with more than 20 reads on average, regulate mammary gland plasticity and may be related to the observed differences in milk production between Holstein and Montbéliarde, which are two breeds with different mammogenic potential. Some of the 17 miRNAs could potentially target mRNAs involved in signaling pathways (such as mTOR) and in lipid metabolism, thereby suggesting that they could influence milk composition. In conclusion, we showed differences in mammary gland miRNomes of two dairy bovine breeds. These differences suggest a potential role of miRNAs in mammary gland plasticity and in milk component synthesis related to milk production and composition.
Project description:Profitable milk production in dairy cows requires good reproductive performance. Calving interval is a trait used to measure reproductive efficiency. Here we used a novel lactating Holstein cow model with genetic and phenotypic divergence in calving interval. Cows had similar genetic merit for milk production traits, but either very good genetic merit for fertility (Fert+; n = 7) or very poor genetic merit for fertility (Fert-; n = 6). We sequenced the transcriptome of endometrial biopsies collected on day 7 of the estrous cycle, a crucial stage for establishment of pregnancy. Significant differential expression (FDR adjusted p < 0.1) of 403 genes was found between the Fert+ and Fert- animals. A standard over-representation analysis resulted in 12 lowly populated canonical pathways; however a novel in-depth analysis revealed a core of eighteen genes both differentially expressed and highly abundant. Almost all of these genes were related to actin-related and cytoskeletal components of the endometrium; fundamental cell-structure differences existed between Fert+ and Fert- phenotypes. We also present results indicating inflammation-related genes are down-regulated, and that multiple ion transporters and gated-voltage channels were up-regulated, in Fert+ cows. These three topics are discussed in terms of the uterus and in the context of fertility during early embryo development. Transcriptomic data analysis is complex; we use a method that filters those genes that may be of greatest importance to the tissue at the timepoint of sampling. Our findings support the utility of this approach, as well as highlighting clear physiological differences in our animal model.
Project description:The fertility of dairy cows is challenged during early lactation and better nutritional strategies need to be developed to address this issue. Combined supplementation of folic acid and vitamin B12 improves energy metabolism in the dairy cow during early lactation. Therefore, the present study was undertaken to explore the effects of this supplement on gene expression in granulosa cells from the dominant follicle during the postpartum period. Multiparous Holstein cows received weekly intramuscular injection of 320 mg folic acid and 10 mg vitamin B12 (treated group) beginning 24 (SD 4) d before calving until 56 d after calving, whereas the control group received saline. The urea plasma concentration was significantly decreased during the pre-calving period, and the concentration of both folate and vitamin B12 were increased in treated animals. Milk production and dry matter intake were not significantly different between the two groups. Plasma concentrations of folates and vitamin B12 were increased in vitamin-treated animals. Daily dry matter intake was not significantly different between the 2 groups before (13.5 kg SE 0.5) and after (23.6 kg SE 0.9) calving. Average energy-corrected milk tended to be greater in vitamin-treated cows, 39.7 (SE 1.4) and 38.1 (SE 1.3) kg/d for treated and control cows, respectively. After calving, average plasma concentration of BHBA tended to be lower in cows injected with the vitamin supplement, 0.47 (SE 0.04) vs. 0.55 (SE 0.03) for treated and control cows, respectively. The ovarian follicle ? 12 mm in diameter was collected by ovarian pick-up after estrus synchronization. Recovered follicular fluid volumes were greater in the vitamin-treated group. A microarray platform was used to investigate the impact of treatment on gene expression of granulosa cells. Lower expression of genes involved in the cell cycle and higher expression of genes associated with granulosa cell differentiation prior to ovulation were observed. Selected candidate genes were analyzed by reverse transcription quantitative polymerase chain reaction. Although the effects of intramuscular injections of folic acid and vitamin B12 on lactational performance and metabolic status of animals were limited, Ingenuity Pathway Analysis of gene expression in granulosa cells suggests a stimulation of cell differentiation in vitamin-treated cows, which may be the result of an increase in LH secretion.
Project description:The fertility of dairy cows is challenged during early lactation and better nutritional strategies need to be developed to address this issue. Combined supplementation of folic acid and vitamin B12 improves energy metabolism in the dairy cow during early lactation. Therefore, the present study was undertaken to explore the effects of this supplement on gene expression in granulosa cells from the dominant follicle during the postpartum period. Multiparous Holstein cows received weekly intramuscular injection of 320 mg folic acid and 10 mg vitamin B12 (treated group) beginning 24 (SD 4) d before calving until 56 d after calving, whereas the control group received saline. The urea plasma concentration was significantly decreased during the pre-calving period, and the concentration of both folate and vitamin B12 were increased in treated animals. Milk production and dry matter intake were not significantly different between the two groups. Plasma concentrations of folates and vitamin B12 were increased in vitamin-treated animals. Daily dry matter intake was not significantly different between the 2 groups before (13.5 kg SE 0.5) and after (23.6 kg SE 0.9) calving. Average energy-corrected milk tended to be greater in vitamin-treated cows, 39.7 (SE 1.4) and 38.1 (SE 1.3) kg/d for treated and control cows, respectively. After calving, average plasma concentration of BHBA tended to be lower in cows injected with the vitamin supplement, 0.47 (SE 0.04) vs. 0.55 (SE 0.03) for treated and control cows, respectively. The ovarian follicle ? 12 mm in diameter was collected by ovarian pick-up after estrus synchronization. Recovered follicular fluid volumes were greater in the vitamin-treated group. A microarray platform was used to investigate the impact of treatment on gene expression of granulosa cells. Lower expression of genes involved in the cell cycle and higher expression of genes associated with granulosa cell differentiation prior to ovulation were observed. Selected candidate genes were analyzed by reverse transcription quantitative polymerase chain reaction. Although the effects of intramuscular injections of folic acid and vitamin B12 on lactational performance and metabolic status of animals were limited, Ingenuity Pathway Analysis of gene expression in granulosa cells suggests a stimulation of cell differentiation in vitamin-treated cows, which may be the result of an increase in LH secretion. Two conditions experiment (Control and Treated). Granulosa cells from the 66h post second PGF2alpha injection. Biological replicates: 3 from each time point. Two technical replicates for each comparison (dye-swap).
Project description:Cows were fed a lactation diet at ad libitum intake (n = 6). At 27±3 days in milk, cows were injected with 50 µg of LPS E. coli in one healthy rear mammary quarter. Milk samples were collected just before LPS challenge (LPS-) and 6.5 h after LPS challenge (LPS+) from the same cows. Microarray analysis was performed using customized 8x60K ruminant miRNA microarrays to compare LPS- to LPS+ miRNome. MiRNome comparison between LPS- and LPS+ identified 37 differentially abundant miRNAs (q-value ≤ 0.05)
Project description:Twelve midlactation cows received 4 diets differing in forage-to-concentrate ration (High (HF) versus Low (LF) forage supplemented or not with lipids (HF with whole intact rapeseeds (HF-RS) and LF with sunflower oil (LF-SO)) 12 cows got into 4 groups, each cow was received 4 different diets in a latin square design Green*txt and Red_*txt raw data files contain Cy3 and Cy5 signal intensities, respectively.
Project description:IgG was collected from virgin and pregnant listeria challenged mouse plasma. Polyclonal IgG was analyzed by reduction, alkylation, and LC-MS with top-down MS/MS (HCD and ETD)to look for glycosylation PTMs
Project description:Genetic variation governs protein expression through both transcriptional and post-transcriptional processes. To investigate this relationship, we combined a multiplexed, mass spectrometry-based method for protein quantification with an emerging mouse model harboring extensive genetic variation from 8 founder strains. We collected genome-wide mRNA and protein profiling measurements to link genetic variation to protein expression differences in livers from 192 diversity outcross mice. We observed nearly 3,700 protein-level quantitative trait loci (pQTL) with an equal proportion of proteins regulated directly by their cognate mRNA as uncoupled from their transcript. Our analysis reveals an extensive array of at least five models for genetic variant control of protein abundance including direct protein-to-protein associations that act to achieve stoichiometric balance of functionally related enzymes and subunits of multimeric complexes.