Project description:We adopted a transcriptome-wide microarray analysis approach to determine the extent to which vascular gene expression is altered as a result of juvenile obesity and identify obesity-responsive mRNAs. We examined transcriptional profiles in the left anterior descending coronary artery (LAD), perivascular fat adjacent to the LAD, and descending thoracic aorta between obese (n=5) and lean (n=6) juvenile Ossabaw pigs (age=22 weeks). Obesity was experimentally induced by feeding the animals a high-fat/high fructose corn syrup/high-cholesterol diet for 16 weeks. We found that expression of 189 vascular cell genes in the LAD and expression of 165 genes in the thoracic aorta were altered with juvenile obesity (FDRM-bM-^IM-$10%) with an overlap of only 28 genes between both arteries. Notably, a number of genes found to be markedly up-regulated in the LAD of obese pigs are implicated in atherosclerosis, including ACP5, LYZ, CXCL14, APOE, PLA2G7, LGALS3, SPP1, ITGB2, CYBB, and P2RY12. Furthermore, pathway analysis revealed the induction of pro-inflammatory and pro-oxidant pathways with obesity primarily in the LAD. Gene expression in the LAD perivascular fat was minimally altered with juvenile obesity. Together, we provide new evidence that obesity produces artery-specific changes in pre-translational regulation with a clear up-regulation of pro-atherogenic genes in the LAD. Our data may offer potential viable drug targets and mechanistic insights regarding the molecular precursors involved in the origins of over-nutrition and obesity-associated vascular disease. In particular, our results suggest that the oxLDL-LOX-1-NFM-NM-:B signaling axis may be involved in the early initiation of a juvenile obesity-induced pro-atherogenic coronary artery phenotype. We examined transcriptional profiles in the left anterior descending coronary artery (LAD), perivascular fat adjacent to the LAD, and descending thoracic aorta between obese (n=5) and lean (n=6) juvenile Ossabaw pigs (age= 22 weeks). All three tissue types were taken from each animal, and each was applied to one and only one array array except a single Thoracic aorta (Animal ID 63 because there were not enough arrays), so there were 32 total arrays (11 unique pigs).
Project description:Dnmt1 is an important regulator of tissue development and differentiation. To assess the effects of epithelium Dnmt1 deletion in the developing urogenital sinus (precursor of the urethra and prostate in males), we isolated urogenital sinus epithelial tissue from Dnmt1 deleted mouse embryos and wildtype mouse embryos. The transcriptomes were analyzed by RNA-seq
Project description:The Wilms tumor protein Wt1 is a transcription factor known to play an important role in urogenital development. Mutations in the human Wt1 encoding gene (WT1) lead to several syndromes associated with defective renal and sexual development, namely WAGR-, Denys-Drash-, and Frasier-syndrome. During mammalian embryogenesis, urogenital development starts with formation of the urogenital ridges, consisting of the embryonic kidneys, called mesonephroi, and the gonads. In Wt1 knockout mice, these ridges start to form, degenerate, however, during further embryonic development . Although WT1 and the syndromes associated with its mutation are known for several years, it is still unclear which genes, regulated by Wt1, mediate the whole variety of Wt1's function in gonad and mesonephros development. To obtain a comprehensive view on Wt1-dependent gene activity in developing urogenital ridges, we employed microarray analysis. Comparison of gene expression in Wt1 wild-type and knockout mice, led to the identification of almost 150 genes with differences in expression levels higher than factor three. Keywords: wild-type and knockout comparison
Project description:The gut microbiota is closely associated with digestion, metabolism, immunity, and host health. The imbalance of the microbial community in livestock directly affects their well-being and, consequently, productivity. The composition and diversity of the gut microbiota are influenced not only by host genetics but also by environmental factors such as the microbial complexity of the rearing environment, feeds, and antibiotics. Here, we focus on the comparison of gut microbial communities in miniature pigs developed for xenotransplantation in specific pathogen-free (SPF) and conventional (non-SPF) facilities. To identify the disparities in gut microbial composition and functionality between these two environments, 16S RNA metagenome sequencing was conducted using fecal samples. The results revealed that the non-SPF pigs had higher gut microbiota diversity than the SPF pigs. The genera Streptococcus and Ruminococcus were more abundant in SPF pigs than in non-SPF pigs. Blautia, Bacteroides, and Roseburia were exclusively observed in SPF pigs, whereas Prevotella was exclusively found in non-SPF pigs. Carbohydrate and nucleotide metabolism, as well as environmental information processing, were predicted to be enriched in SPF pigs. In addition, energy and lipid metabolism, along with processes related to genetic information, cellular communication, and diseases, were predicted to be enriched in non-SPF pigs. This study makes an important contribution to elucidating the impact of environments harboring a variety of microorganisms, including pathogens, on the gut microbiota of miniature pigs. Furthermore, we sought to provide foundational data on the characteristics of the gut microbiota in genetically modified pigs, which serve as source animals for xenotransplantation.
Project description:The goal of this experiment was to determine if testosterone (T) and estradiol-17 beta (E2) induced gene expression in mouse urogenital sinus mesenchymal (UGM) cells. When UGM grown in culture was treated with T+E2, numerous genes were increased and decreased compared to untreated controls. Quantitative RT-PCR was used to verify results of a subset of genes.