Project description:Different lactation stages have marked influence on milk yield, milk constituents and nourishment of the neonates. However, the differential gene expression during different lactation stages in Bosindicus has not been investigated so far. In this study, we carried out high-resolution mass spectrometry-based quantitative proteomics of bovine whey at early, mid and late lactation stages of MalnadGidda (Bosindicus) cows. Using TMT-based quantitative proteomics, we compared the bovine whey proteins on progressive lactation stages of Indian breed, MalnadGidda(Bosindicus). LC-MS/MS analysis of whey peptides from early, mid and late lactation stages resulted in the generation of 420,092 MS/MS spectra and 50,800 peptide spectrum matches, which led to the identification of 4,450 peptides corresponding to 725 proteins. Out of which, 440 proteins were differentially expressed (≥1.5-fold). Gene Ontology studies showed that proteins that regulatemilk composition and mammary growth associated proteins are abundantly expressed during peak lactation stages. Whereas, proteins related to pregnancy and mammary involution are expressed high in late and mid lactation stages indicating the physiological changes in the maternal system of bovine during drying period. Detection of progestagen associated endometrial protein; an immune protein seen in the fetomaternal interface and other pregnancy associated proteins at mid lactation suggest a candidate biomarker for the early pregnancy diagnosis. These results are overlapping with the previous findings addressed in milk from exotic breeds. We strongly believe that this preliminary investigation on differential proteome in milk whey over the course of lactation of indigenous cattle could answer many unsolved questions in lactation biology.
Project description:The transition between pregnancy and lactation is a major physiological change that dairy cows must contend with. Complex systemic and local processes involving gluconeogenesis, energy balance, utilisation of body reserves, insulin resistance and involution of the uterus can have an effect on animal health and farm profitability. Here we used an established Holstein cow model of fertility that displayed genetic and phenotypic divergence in calving interval, a trait used to define reproductive performance using a national breeding index in Ireland. Cows had similar genetic merit for milk production traits, but either very good genetic merit for fertility (‘Fert+’; n = 8) or very poor genetic merit for fertility (‘Fert-‘; n = 8). We investigated three distinct time-points, late pregnancy, early lactation and mid lactation (-18, 1 and 147 days on average with day 0 being birth), using RNA sequencing from both liver and muscle tissue biopsies and conducting a differential expression (DE) analysis. We found 807 and 815 unique genes to be DE in at least one time-point in liver and muscle respectively, of which 79% and 83% were only found in a single time-point; 40 and 41 genes were found DE at every time-point indicating possibly systemic or chronic dysregulation. Functional annotation resulted in evidence for two major physiological processes: immune and inflammation, and metabolic, lipid and carbohydrate-binding. These processes indicate areas of previous interest as well as specific systems that appear differentially regulated, and point towards interesting avenues of further research in a broad and complex field.
Project description:Infertility is a disease that affects humans and cattle in similar ways. The resemblance includes complex genetic architecture, multiple etiology, low heritability of fertility related traits in females, and the frequency in the female population. Here, we used cattle as a biomedical model to test the hypothesis that gene expression profiles of protein-coding genes expressed in peripheral white blood cells (PWBCs), and circulating micro RNAs in plasma, are associated with female fertility, measured by pregnancy outcome. We drew blood samples from 17 female calves on the day of artificial insemination and analyzed transcript abundance for 10496 genes in PWBCs and 290 circulating micro RNAs. The females were later classified as pregnant to artificial insemination, pregnant to natural breeding or not pregnant. We identified 1860 genes producing significant differential coexpression (eFDR<0.002) based on pregnancy outcome. Additionally, 237 micro RNAs and 2274 genes in PWBCs presented differential coexpression based on pregnancy outcome. Furthermore, using a machine learning prediction algorithm we detected a subset of genes whose abundance could be used for blind categorization of pregnancy outcome. Our results provide strong evidence that bloodborne transcript abundance is highly associated with fertility in females.
Project description:The liver of dairy cows naturally displays a series of metabolic adaptation during the periparturient period in response to the increasing nutrient requirement of lactation. The hepatic adaptation is partly regulated by insulin resistance and it is affected by the prepartal energy intake level of cows. We aimed to investigate the metabolic changes in the liver of dairy cows during the periparturient at gene expression level and to study the effect of prepartal energy level on the metabolic adaptation at gene expression level.B13:N13
Project description:The mammary gland undergoes extensive remodeling between the begin- ning of pregnancy and lactation; this involves cellular processes including cell proliferation, differentiation, and apoptosis, all of which are under the control of numerous regulators. To unravel the role played by miRNA, we describe here 47 new ovine miRNA cloned from mammary gland in early pregnancy displaying strong similari- ties with those already identified in the cow, human, or mouse. A microarray study of miRNA variations in the adult ovine mammary gland during pregnancy and lactation showed that 100 miRNA are regulated according to three principal patterns of expression: a de- crease in early pregnancy, a peak at midpregnancy, or an increase throughout late pregnancy and lactation. One miRNA displaying each pattern (miR-21, miR-205, and miR-200b) was analyzed by qRT- PCR. Variations in expression were confirmed for all three miRNA. Using in situ hybridization, we detected both miR-21 and miR-200 in luminal mammary epithelial cells when expressed, whereas miR-205 was expressed in basal cells during the first half of pregnancy and then in luminal cells during the second half. We therefore conclude that miR-21 is strongly expressed in the luminal cells of the normal mammary gland during early pregnancy when extensive cell prolif- eration occurs. In addition, we show that miR-205 and miR-200 are coexpressed in luminal cells, but only during the second half of pregnancy. These two miRNA may cooperate to maintain epithelial status by repressing an EMT-like program, to achieve and preserve the secretory phenotype of mammary epithelial cells. 5 samples for sheep and 5 samples for mouse