Project description:The mammalian oviduct is a dynamic organ and the venue of important events leading to the establishment of pregnancy. Oviductal epithelial cells are involved in direct contact and specific interactions with different self and non-self entities (oocyte, sperm, etc.).The aim of this study was to determine if the oviduct is able to distinguish between different types of non-self-entities. We used an in vitro model to determine genes altered in oviductal epithelial cell (OPEC) in response to the presence of spermatozoa, mammalian somatic cells (red blood cell) or bacteria. We hypothesised that alteration of OPEC expression profile in response to boar spermatozoa (friendly non-self entity), red blood cells (non-pathogenic, non-self entity) and bacteria (pathogenic non-self entity) is indicative of recognition of self and non-self entities by these cells as well as recognition of pathogenic from non-pathogenic non-self entities.
Project description:The mammalian oviduct is a dynamic organ and the venue of important events leading to the establishment of pregnancy. Oviductal epithelial cells are involved in direct contact and specific interactions with different self and non-self entities (oocyte, sperm, etc.).The aim of this study was to determine if the oviduct is able to distinguish between different types of non-self-entities. We used an in vitro model to determine genes altered in oviductal epithelial cell (OPEC) in response to the presence of spermatozoa, mammalian somatic cells (red blood cell) or bacteria. We hypothesised that alteration of OPEC expression profile in response to boar spermatozoa (friendly non-self entity), red blood cells (non-pathogenic, non-self entity) and bacteria (pathogenic non-self entity) is indicative of recognition of self and non-self entities by these cells as well as recognition of pathogenic from non-pathogenic non-self entities. Oviductal epithelial cells (OPEC) were co-incubated in the presence of (i) bacteria (1 x 10˄8 bacteria), (ii) sperm (1 X 10 ˄ 6 spermatozoa/ml) or (iii) red blood cell (5 x 10 ˄ 6 red blood cell/ml) for 24 hours in HAM-12 media. Three biological replicates were performed for each group and a total of 12 Affymetrix Porcine arrays were used to identify the expression profiles of OPEC alone (3 arrays) or in the presence of (i) bacteria (3 arrays), (ii) sperm (3 arrays) or (iii) red blood cells (3 arrays).
Project description:Large White and Meishan pigs were either non-treated or injected with mammalian 1-24 ACTH (Immediate Synachten, Novartis France) at the dose of 250 µg per animal. Pigs were sacrificed either immediately after capture from their home cage (non-treated animals) or 1 hour following ACTH injection. Adrenal glands were immediately collected from pigs and frozen on dry ice and then stored at -80°C until RNA isolation. Keywords: stress response, adrenal, gene expression, pig
Project description:Large White and Meishan pigs were either non-treated or injected with mammalian 1-24 ACTH (Immediate Synachten, Novartis France) at the dose of 250 µg per animal. Pigs were sacrificed either immediately after capture from their home cage (non-treated animals) or 1 hour following ACTH injection. Adrenal glands were immediately collected from pigs and frozen on dry ice and then stored at -80°C until RNA isolation. Keywords: stress response, adrenal, gene expression, pig 47 samples
Project description:Regulatory Mechanisms of Atrial Remodeling of Mitral Regurgitation Pigs This study enrolled 6 pigs (age: 18 months) and divided into three groups: mitral regurgitation pigs (MR) (n = 2; 2 males sacrificed 12 months after surgery), MR pigs treated with valsartan (MRV) (n = 2; 2 males age-matched to MR sacrificed 12 months after surgery), and normal control pigs (NC) (n = 2; 2 males age-matched to MR pigs). Valsartan (3.43 mg/kg/day), a type I angiotensin II receptor blocker, was administered from one week before surgery and then daily after surgery in the MRV group. We sought to systemically elucidate critical differences in the alteration of RNA expression pattern between the atrial myocardium of pigs with and without MR, and between the atrial myocardium of MR pigs with and without valsartan using high-density oligonucleotide microarrays and functional network enrichment analysis.
Project description:BACKGROUND:In animal breeding, identification of causative genetic variants is of major importance and high economical value. Usually, the number of candidate variants exceeds the number of variants that can be validated. One way of prioritizing probable candidates is by evaluating their potential to have a deleterious effect, e.g. by predicting their consequence. Due to experimental difficulties to evaluate variants that do not cause an amino-acid substitution, other prioritization methods are needed. For human genomes, the prediction of deleterious genomic variants has taken a step forward with the introduction of the combined annotation dependent depletion (CADD) method. In theory, this approach can be applied to any species. Here, we present pCADD (p for pig), a model to score single nucleotide variants (SNVs) in pig genomes. RESULTS:To evaluate whether pCADD captures sites with biological meaning, we used transcripts from miRNAs and introns, sequences from genes that are specific for a particular tissue, and the different sites of codons, to test how well pCADD scores differentiate between functional and non-functional elements. Furthermore, we conducted an assessment of examples of non-coding and coding SNVs, which are causal for changes in phenotypes. Our results show that pCADD scores discriminate between functional and non-functional sequences and prioritize functional SNVs, and that pCADD is able to score the different positions in a codon relative to their redundancy. Taken together, these results indicate that based on pCADD scores, regions with biological relevance can be identified and distinguished according to their rate of adaptation. CONCLUSIONS:We present the ability of pCADD to prioritize SNVs in the pig genome with respect to their putative deleteriousness, in accordance to the biological significance of the region in which they are located. We created scores for all possible SNVs, coding and non-coding, for all autosomes and the X chromosome of the pig reference sequence Sscrofa11.1, proposing a toolbox to prioritize variants and evaluate sequences to highlight new sites of interest to explain biological functions that are relevant to animal breeding.
Project description:Gene expression of characteristic chondrogenic markers and miRNA expression were analyzed in cells cultured in differentiation medium and significant differences were found between gelation/PRP microgels and those containing only pure gelatin. We used microarrays to detail the miRNA expression in studied cell cultures for identification the expression of miRNA and study the up- and down-regulated miRNA associated.
Project description:Maternal exposure to estrogens can induce long-term adverse effects in the offspring. This may be mediated through alterations in the endometrium affecting embryo-maternal communication as early as the preimplantational phase. Thus, we analyzed the effects of gestational estradiol-17β (E2) exposure on the endometrium. Two distinct low doses and a high dose (0.05, 10 and 1000 µg E2/kg body weight daily, respectively) were orally applied to sows from insemination until sampling at day 10 of pregnancy and compared to carrier-treated controls. RNA-sequencing revealed a dose-dependent increase of 14, 17 and 27 differentially expressed genes (DEG), respectively. Overall, the maternal E2 treatment perturbed gene expression of the endometrium, potentially altering the uterine histotroph.