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: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:Weaning is a very critical period for piglets, typically accompanied by lower feed intake, weight loss after weaning and increased mortality. At weaning, piglets are exposed to many stressors, such as loss of mothering, mixing with other litters, end of lactational immunity, and a change in their environment and gut microbiota. After weaning, morphological and histological changes occur in the small intestine of piglets producing a rapid change of feeding regime which is critical for the immature digestive system. Sixteen female piglets were weaned to assess the effect of sorbic acid supplementation on the small intestine tissue transcriptome. At weaning day (T0), 4 piglets were sacrified and tissue samples collected. The remaining 12 piglets were weighted and randomly assigned to different post weaning (T5) diets. Diet A (n=6) contained 5 g/kg of sorbic acid. Diet B (n=6) is the same as Standard diet. Total RNA was isolated from ileum samples to be analyzed using the a CombiMatrix CustomArrayTM 90K platform . Even though diet had no detectable effect during the first 5 days after weaning, outcomes from this study highlighted some of the response mechanisms to the stress of weaning occurring in the piglet gut. A total of 205 differentially expressed genes were used for functional analysis using bioinformatics through BLAST2GO, Ingenuity Pathway Analysis 8.0, and the Dynamic Impact Aproach (DIA). Bioinformatics analysis revealed that Apoptosis, RIG-I-like and NOD-like receptor signaling were altered as a result of weaning. Results suggest that immune and inflammatory responses were activated and likely are a cause of small intestine atrophy as revealed by a decrease in villus height and villus/crypt ratio. Keywords: weaning, gut, gene expression, sorbic acid, microarray analysis
Project description:To obtain an overview of the transcriptome landscape in developing pig skeletal muscle, 81 high-quality transcriptome libraries that covered 27 developmental stages (3 biological replicates per stage) in pig skeletal muscle were produced by strand-specific rRNA-depleted total RNA sequencing (RNA-seq). We generated 8.59 billion paired-end reads (150 bp × 2) covering 1.24 Tb of sequence for RNA-seq.
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