Project description:In sheep, the innate immune response of mammary epithelial cells (MECs) plays a central role in combating mastitis, yet our understanding of their resistance mechanisms remains limited. This study aimed to elucidate the gene expression profiles of ovine MECs following in vitro stimulation with Staphylococcus aureus (S. aureus) using RNA-Seq technology. Bioinformatics analysis identified a total of 175 differentially expressed genes (DEGs), including 172 up-regulated and 3 down-regulated genes in the stimulated group compared to the non-stimulated control group. Gene ontology annotation and functional pathway analysis indicated that these DEGs are primarily involved in ribosomal functions, which are essential for protein synthesis and first target of pathogens, as well as in immune response dysregulations, infection, phagocytosis, and bacterial invasion of epithelial cells. Validation via quantitative real-time PCR (qRT-PCR) confirmed the RNA-Seq results. These findings significantly contribute to the understanding of how ovine MECs respond to S. aureus stimulation, providing a foundation for further research, particularly regarding the immune defense mechanisms, strategies and implications in dairy industry.
Project description:Comparative expression profiles between unstimulated ovine keratinocytes and keratinocytes stimulated with whole mite antigen and whole mite wash in vitro, over a time course of 1, to 48 hours, using the RIGUA custom array (Watkins et al., 2008) and real-time RT-PCR
Project description:A single nucleotide polymorphism (SNP) in the human glucocorticoid receptor (GR), N363S, has been the focus of several clinical studies, and some epidemiological data link this SNP to increased glucocorticoid sensitivity, coronary artery disease and increased body mass index (BMI). However, molecular studies in vitro using reporter gene expression systems have failed to define a link between this polymorphism and altered glucocorticoid receptor function. To address the biological relevancy of N363S in glucocorticoid receptor mechanisms and function, we established stable U-2 OS (human osteosarcoma) cell lines expressing wild type hGR or N363S using a tetracycline-regulated expression system. Functional assays with reporter gene systems revealed only minor differences between the wild type hGR and N363S receptors under a variety of conditions that probe for GR function. However, examination of this polymorphism by human gene microarray analysis showed, for the first time, that there are significant differences between wild type hGR and the N363S SNP in their ability to selectively regulate gene expression. Several of these genes may define the link between the N363S SNP and human disease. Keywords: human glucocorticoid receptor, N363S single nucleotide polymorphism, microarray gene analysis
Project description:Uncovering dark mass in population proteomics: Pan-analysis of Single Amino-acid Polymorphism (Pan-SAP) revealed quantitative relevance with cognition and aging
Project description:PPARG ChIP seq analysis was conducted to determine genes bound by and potentially regulated by PPARG in the developing ovine conceptus. Determination of gene regulation by prostaglandins through PPARG helps to improve our understanding of early pregnancy events and provides a basis for strategies to improve fertility and reproductive efficiency in ruminants.
