Project description:It is well known that host-microbes and immunity interactions are influenced by dietary patterns, as well as daily environmental light-dark (LD) cycles that entrain circadian rhythms in the host. Emerging data has highlighted the importance of diet patterns and timing on the interaction among circadian rhythms, gut microbiome, and immunity, however, their impacts on LD cycles are less reported. Therefore, we aim to study how LD cycles regulate the homeostatic crosstalk between gut microbiome, hypothalamic and hepatic circadian clock oscillations and immunity. We hypothesized that different environmental LD cycles: (1) constant darkness, LD0/24; (2) short light, LD8/16; (3) normal LD cycle, LD12/12; (4) long light, LD16/8; and (5) constant light, LD24/0, may affect immunity and metabolism to varying degrees. Therefore, 240 mice were managed with chow diets (CD) and antibiotics treatments (ABX) under five different LD cycles for 42 days. The colonic (co) and cecum (ce) contents were obtained for studying their impacts on gut microbiome using 16S rRNA sequencing.
Project description:It has been widely recognized that the microbiota has the capacity to shape host gene expression and physiological functions. However, there remains a paucity of comprehensive study revealing host transcriptional landscape regulated by the microbiota. Here, we comprehensively examined mRNA landscapes in mouse tissues (brain and cecum) from specific pathogen free (SPF) and germ-free mouse (GF) using Nanopore direct RNA sequencing. Our results show that the microbiome has global influence on host’s RNA modifications (m6A, m5C, Ψ), isoform generation, poly(A) tail length (PAL), and transcript abundance in both brain and cecum tissues. Moreover, the microbiome exerts tissue-specific effects on various post-transcriptional regulatory processes. In addition, the microbiome impacts the coordination of multiple RNA modifications in host brain and cecum tissues. In conclusion, we establish the relationship between microbial regulation and gene expression, our results help the understanding of the mechanisms by which the microbiome reprograms host gene expression.
Project description:It has been widely recognized that the microbiota has the capacity to shape host gene expression and physiological functions. However, there remains a paucity of comprehensive study revealing host transcriptional landscape regulated by the microbiota. Here, we comprehensively examined mRNA landscapes in mouse tissues (brain and cecum) from specific pathogen free (SPF) and germ-free mouse (GF) using Nanopore direct RNA sequencing. Our results show that the microbiome has global influence on host’s RNA modifications (m6A, m5C, Ψ), isoform generation, poly(A) tail length (PAL), and transcript abundance in both brain and cecum tissues. Moreover, the microbiome exerts tissue-specific effects on various post-transcriptional regulatory processes. In addition, the microbiome impacts the coordination of multiple RNA modifications in host brain and cecum tissues. In conclusion, we establish the relationship between microbial regulation and gene expression, our results help the understanding of the mechanisms by which the microbiome reprograms host gene expression.
Project description:Cruciferous vegetables and their derived compounds, such as glucosinolates, have demonstrated anti-inflammatory properties in animal models. In this study, broccoli-supplemented diet induced changes in the gut microbiome and its role in host and bacterial gene expression were examined in mice following consumption of the Total Western Diet (TWD), which is based on NHANES data and represents the composition of a typical American diet. C57BL/6 male mice were fed a TWD for six weeks followed by a supplementation of 0, 0.5, 1 or 2.5% broccoli powder for three weeks. Microbial communities from cecal contents were taxonomically profiled using 16S and metagenomics sequencing, and metatranscriptomics was used to assess functionality of bacterial species. Cecum tissues were also analyzed for host transcriptomics.
Project description:Cruciferous vegetables and their derived compounds, such as glucosinolates, have demonstrated anti-inflammatory properties in animal models. In this study, broccoli-supplemented diet induced changes in the gut microbiome and its role in host and bacterial gene expression were examined in mice following consumption of the Total Western Diet (TWD), which is based on NHANES data and represents the composition of a typical American diet. C57BL/6 male mice were fed a TWD for six weeks followed by a supplementation of 0, 0.5, 1 or 2.5% broccoli powder for three weeks. Microbial communities from cecal contents were taxonomically profiled using 16S and metagenomics sequencing, and metatranscriptomics was used to assess functionality of bacterial species. Cecum tissues were also analyzed for host transcriptomics.
Project description:Background- Resistant starch is a prebiotic metabolized by the gut bacteria. It has been shown to attenuate chronic kidney disease (CKD) progression in rats. Previous studies employed taxonomic analysis using 16S rRNA sequencing and untargeted metabolomics profiling. Here we expand these studies by metaproteomics, gaining new insight into the host-microbiome interaction. Methods- Differences between cecum contents in CKD rats fed a diet containing resistant starch with those fed a diet containing digestible starch were examined by comparative metaproteomics analysis. Taxonomic information was obtained using unique protein sequences. Our methodology results in quantitative data covering both host and bacterial proteins. Results - 5,834 proteins were quantified, with 947 proteins originating from the host organism. Taxonomic information derived from metaproteomics data surpassed previous 16S RNA analysis, and reached species resolutions for moderately abundant taxonomic groups. In particular, the Ruminococcaceae family becomes well resolved – with butyrate producers and amylolytic species such as R. bromii clearly visible and significantly higher while fibrolytic species such as R. flavefaciens are significantly lower with resistant starch feeding. The observed changes in protein patterns are consistent with fiber-associated improvement in CKD phenotype. Several known host CKD-associated proteins and biomarkers of impaired kidney function were significantly reduced with resistant starch supplementation. Conclusions- Metaproteomics analysis of cecum contents of CKD rats with and without resistant starch supplementation reveals changes within gut microbiota at unprecedented resolution, providing both functional and taxonomic information. Proteins and organisms differentially abundant with RS supplementation point toward a shift from mucin degraders to butyrate producers.