Project description:Gut microbiome research is rapidly moving towards the functional characterization of the microbiota by means of shotgun meta-omics. Here, we selected a cohort of healthy subjects from an indigenous and monitored Sardinian population to analyze their gut microbiota using both shotgun metagenomics and shotgun metaproteomics. We found a considerable divergence between genetic potential and functional activity of the human healthy gut microbiota, in spite of a quite comparable taxonomic structure revealed by the two approaches. Investigation of inter-individual variability of taxonomic features revealed Bacteroides and Akkermansia as remarkably conserved and variable in abundance within the population, respectively. Firmicutes-driven butyrogenesis (mainly due to Faecalibacterium spp.) was shown to be the functional activity with the higher expression rate and the lower inter-individual variability in the study cohort, highlighting the key importance of the biosynthesis of this microbial by-product for the gut homeostasis. The taxon-specific contribution to functional activities and metabolic tasks was also examined, giving insights into the peculiar role of several gut microbiota members in carbohydrate metabolism (including polysaccharide degradation, glycan transport, glycolysis and short-chain fatty acid production). In conclusion, our results provide useful indications regarding the main functions actively exerted by the gut microbiota members of a healthy human cohort, and support metaproteomics as a valuable approach to investigate the functional role of the gut microbiota in health and disease.
Project description:Hibernation is an energy-saving strategy adopted by a wide range of mammals to survive highly seasonal or unpredictable environments. Arctic ground squirrels living in Alaska provide an extreme example, with 6-9 months long hibernation seasons when body temperature alternates between levels near 0 C during torpor and 37 C during arousal episodes. Heat production during hibernation is provided, in part, by non-shivering thermogenesis that occurs in large deposits of brown adipose tissue (BAT). BAT is active at tissue temperatures from 0 to 37 C during rewarming and continuously at near 0 C during torpor in subfreezing conditions. Despite its crucial role in hibernation, the global gene expression patterns in BAT during hibernation compared to the non-hibernation season remain largely unknown. We report a large-scale study of differential gene expression in BAT between winter hibernating and summer active arctic ground squirrels using mouse microarrays. Selected differentially expressed genes identified on the arrays were validated by quantitative real-time PCR using ground squirrel specific primers. Our results show that the mRNA levels of the genes involved in nearly every step of the biochemical pathway leading to non-shivering thermogenesis are significantly increased in BAT during hibernation, whereas those of genes involved in protein biosynthesis are significantly decreased compared to the summer active animals in August. The differentially expressed genes also include those involved in adipose differentiation, substrate transport, and structure remodeling, which may enhance thermogenesis at low tissue temperatures in BAT. Keywords: hibernating animals vs. summer active animals
Project description:miRNAs are 19-25 nucleotides long small RNAs now well-known for their regulatory roles in the development and diseases through post-transcriptional and translational controls in a wide range of species. Mammalian hibernation is a physiological process involving dramatic metabolic suppression and cellular reorganization, during which miRNAs may play an important role. We systematically analyzed the miRNAs in the liver of an extreme hibernating species, arctic ground squirrels (Spermophilus parryii), during two stages of hibernation compared to non-hibernating animals by massively parallel Illumina sequencing technology. We identified more than 200 ground squirrel miRNAs including novel miRNAs specific to ground squirrel and a fast-evolving miRNA cluster that also showed significant differential expression during hibernation. Integrating with Agilent miRNA microarray and Real-time PCR results, we identified that mir-211, mir-378, mir-184, mir-200a, and mir-320 were significantly under-expressed during hibernation, whereas mir-144, mir-486, mir-451, mir-142-5p, and mir-1 were over-expressed. Analyses of the their target genes suggested that these miRNAs could play an important role to suppress tumor progression and cell growth during hibernation. Investigation of microRNA changes in arctic ground squirrel livers during Early Arousal(EA), Late Topor(LT), and Post-Reproduction(PR) stages.
Project description:miRNAs are 19-25 nucleotides long small RNAs now well-known for their regulatory roles in the development and diseases through post-transcriptional and translational controls in a wide range of species. Mammalian hibernation is a physiological process involving dramatic metabolic suppression and cellular reorganization, during which miRNAs may play an important role. We systematically analyzed the miRNAs in the liver of an extreme hibernating species, arctic ground squirrels (Spermophilus parryii), during two stages of hibernation compared to non-hibernating animals by massively parallel Illumina sequencing technology. We identified more than 200 ground squirrel miRNAs including novel miRNAs specific to ground squirrel and a fast-evolving miRNA cluster that also showed significant differential expression during hibernation. Integrating with Agilent miRNA microarray and Real-time PCR results, we identified that mir-211, mir-378, mir-184, mir-200a, and mir-320 were significantly under-expressed during hibernation, whereas mir-144, mir-486, mir-451, mir-142-5p, and mir-1 were over-expressed. Analyses of the their target genes suggested that these miRNAs could play an important role to suppress tumor progression and cell growth during hibernation. Three total RNA pools from arctic ground squirrel livers in Early Arousal(EA), Late Topor(LT), and Post-Reproduction(PR) stages were hybridized to three Agilent mouse miRNA microarrays.
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:Mammalian hibernators display phenotypes similar to physiological conditions in non-hibernating species under conditions of calorie restriction and fasting, hypoxia, hypothermia, ischemia-reperfusion, and sleep. However, whether or how similarities are also reflected on molecular and genetic levels is unclear. We identified molecular signatures of torpor and arousal in hibernation using a new custom-designed cDNA microarray for the arctic ground squirrel (Urocitellus parryii,) and compared them to molecular signatures of selected phenotypes in mouse. Our results show that differential gene expression related to metabolism during torpor is closely related to that during calorie restriction and hypoxia. PPARM-NM-1 is crucial for metabolic remodeling in hibernation. Genes related to the sleep-wake cycle and temperature response genes induced by hypothermia follow the same expression changes as in torpor-arousal cycle. Increased fatty acid metabolism might contribute to the protection against ischemia-reperfusion injury during hibernation. Further, by comparing with thousands of pharmacological signatures, we identified drugs that may induce similar expression patterns in human cell lines as during hibernation. Arctic ground squirrels sampled during winter hibernation were compared with the animals sampled during summer. Liver was hybridized on a custom 9,600 probes nylon membrane microarray platform. Four squirrels in early torpor, five in late torpor, four in early arousal, four in late arousal, and seven in summer active were studied in experiments.