Project description:The aim of the current study was to characterize the differential cellular and exosomal miRNAs during inflammation or high fat diet-induced obesity in mice. Mesenteric adipose tissue (MAT) and abdominal aorta (AA) from mice fed a normal chow diet (NCD) or a high fat diet (HFD) were harvested for miRNA profiling. MAT-derived adipocytes (MAT-Ad) challenged with either lipopolysaccharide (LPS, 1 µg/ml) or PBS were harvested for miRNA profiling. Meanwhile, miRNAs encapsulated in MAT-Ad-derived exosomes (MAT-Ad-EX) were also analyzed. Hierarchical clustering analysis performed on most significantly regulated miRNAs (HFD vs NCD in tissues; LPS challenge vs PBS in the cells) showed a set of miRNAs that are differentially expressed in obese versus lean MAT or AA tissues, and in LPS-challenged versus PBS-treated MAT-Ads. The dysregulated of miRNAs in MAT-Ad-EX was also generated and hierarchically clustered, induced by prolonged exposure to microbial product.
Project description:The vertebrate skeleton is mostly composed of three specific cell types: immature chondrocytes (IMM), mature (hypertrophic) chondrocytes (MAT), and osteoblasts (OST). These three cell types are distinct, but they also share the expression of many genes. This overlapping gene expression can be attributed to two transcription factors, SOX9 and RUNX2, which operate near the top of hierarchy of the gene regulatory network (GRN) underlying IMM, MAT, and OST. Sox9 drives IMM differentiation, whereas Runx2 regulates OST differentiation. Importantly, MAT do not form without the function of either Sox9 or Runx2, but little is known about mechanisms of GRN regulation in MAT. During MAT differentiation, the expression of Runx2 increases, and many genes regulated by this transcription such as Spp1, Mef2c, Ibsp, and Alpl are activated. To understand regulatory control of gene expression in mature chondrocytes, ChIP-seq experiments were performed using the mouse chondrogenic cell line ATDC5. These experiments identified in vitro RUNX2 binding sites at different stages of chondrogenesis. RUNX2 appeared to bind in most genes enriched in MAT at both day 3 of differentiation. The ChIP-seq analyses presented here verified the molecular mechanisms predicted here to regulate transcription of the many genomic loci in MAT, proving more insight into regulatory control during cartilage maturation.
Project description:Neurospora tetrasperma is a pseudohomothallic filamentous ascomycete with a large (~ 7 Mbp) region of suppressed recombination surrounding its mating-type (mat) locus. The suppressed recombination has lead to sequence divergence between the two mating-type chromosomes of wild-type heterokaryotic strains, while the remaining genome is largely homoallelic. In this study, we use microarray technology to manifest expression divergence linked to mating type in N. tetrasperma. N. tetrasperma and N. crassa, were grown on agar regimes inducing sexual growth (Synthetic Crossing medium) and vegetative growth (Vogel's Medium), respectively. [SC]: Neurospora tetrasperma mat-A FGSC#1270; mat-a FGSC#1271; Mat-A FGSC#9033; mat-a FGSC#9034; N. crassa mat-A FGSC#2489 and mat-a FGSC 4200: Synthetic Crossing medium was used as a nutrient regime before sampling and processing [Veg]: Neurospora tetrasperma mat-A FGSC#1270; mat-a FGSC#1271; Mat-A FGSC#9033; mat-a FGSC#9034; N. crassa mat-A FGSC#2489 and mat-a FGSC 4200: Vogel's Medium (Vegetative Medium) was used as a nutrient regime before sampling and processing
Project description:Species within the human pathogenic Cryptococcus species complex are major threats to public health, causing about one million infections globally each year. Cryptococcus amylolentus is the most closely known related species of the pathogenic Cryptococcus species complex, and it is non-pathogenic. Additionally, while pathogenic Cryptococcus species have bipolar mating systems with a single large MAT locus that represents a derived state in Basidiomycetes, C. amylolentus has a tetrapolar mating system with two MAT loci (P/R and HD) located on different chromosomes. Thus, studying C. amylolentus will shed light on the origin and evolution of pathogenesis, as well as the transition from tetrapolar to bipolar mating systems in the pathogenic Cryptococcus species. In this study, we sequenced, assembled, and annotated the genomes of two C. amylolentus isolates, CBS6039 and CBS6273, which are sexual and interfertile. Genome comparison between the two C. amylolentus isolates identified the boundaries and the complete gene contents of the P/R and HD MAT loci. Also, bioinformatics and ChIP-seq analyses showed that C. amylolentus has regional centromeres that are enriched with species-specific transposable and repetitive elements, similar to the centromeric structures in the pathogenic Cryptococcus species. Additionally, we found that while neither of the P/R and HD loci in C. amylolentus is physically linked to its centromere, both MAT loci exhibit centromere linkage in meiosis, suggesting the presence of recombinational suppressors and/or epistatic gene interactions in the inter MAT-CEN regions. Furthermore, genomic comparison between C. amylolentus and pathogenic Cryptococcus species provides evidence that chromosomal rearrangements mediated by intercentromeric recombination have occurred after the two lineages split from their common ancestor. We propose a model in which the evolution of the bipolar mating system was initiated by an ectopic recombination event mediated by repetitive elements located within the centromeric regions and shared between chromosomes. This translocation brought the P/R and HD loci onto the same chromosome, and was followed by chromosomal rearrangements that resulted in the two MAT loci becoming physically linked and eventually fused to form the single contiguous MAT locus that is now extant in the pathogenic Cryptococcus species.
Project description:This SuperSeries is composed of the following subset Series: GSE36154: Expression analysis of HFF cells in G0 phase [MAT] GSE36155: Expression analysis of HFF cells in G1 phase [MAT] GSE36156: Differential expression analysis of HFF cells in G0 phase compared to cells in G1 [MAT] Refer to individual Series
Project description:We performed repeated endoscopic colon biopsies of C57BL/6J mice weekly and found that the mesenteric adipose tissue (MAT) was enlarged and it wrapped around the colon at wound sites after 4 cycles of biopsy. On histological view, fibrous bands extended from the intestine into the adjacent MAT, mimicking the fibrosis feature in Crohn’s disease. To investigate the gene signature in the MAT, laser capture microdissection was performed to collect the MAT and associated fibrosis in the repeated biopsy mice and controls. Agilent Microarray was used for the transcriptomic analysis.
Project description:Abstract BACKGROUND: Mating-type loci in yeasts and ascomycotan filamentous fungi (Pezizomycotina) encode master transcriptional factors that play a critical role in sexual development. Genome-wide analyses of mating-type-specification circuits and mating-type target genes are available in Saccharomyces cerevisiae and Schizosaccharomyces pombe; however, no such analyses have been performed in heterothallic (self-incompatible) Pezizomycotina. The heterothallic fungus Podospora anserina serves as a model for understanding the basic features of mating-type control. Its mat+ and mat- mating types are determined by dissimilar allelic sequences. The mat- sequence contains three genes, designated FMR1, SMR1 and SMR2, while the mat+ sequence contains one gene, FPR1. FMR1 and FPR1 are the major regulators of fertilization, and this study presents a genome-wide view of their target genes and analyzes their target gene regulation. METHODOLOGY/PRINCIPAL FINDINGS: The transcriptomic profiles of the mat+ and mat- strains revealed 157 differentially transcribed genes, and transcriptomic analysis of fmr1(-) and fpr1(-) mutant strains was used to determine the regulatory actions exerted by FMR1 and FPR1 on these differentially transcribed genes. All possible combinations of transcription repression and/or activation by FMR1 and/or FPR1 were observed. Furthermore, 10 additional mating-type target genes were identified that were up- or down-regulated to the same level in mat+ and mat- strains. Of the 167 genes identified, 32 genes were selected for deletion, which resulted in the identification of two genes essential for the sexual cycle. Interspecies comparisons of mating-type target genes revealed significant numbers of orthologous pairs, although transcriptional profiles were not conserved between species. CONCLUSIONS/SIGNIFICANCE: This study represents the first comprehensive genome-wide analysis of mating-type direct and indirect target genes in a heterothallic filamentous fungus. Mating-type transcription factors have many more target genes than are found in yeasts and exert a much greater diversity of regulatory actions on target genes, most of which are not directly related to mating.
Project description:Adipose tissue (AT) contains mesenchymal stromal cells (MSC) in stages of commitment to becoming specialized tissue cells, including adipocytes and fibroblasts, and immune cells which support tissue homeostasis. How MSC and immune cells interact during infection is poorly understood. We show that during intestinal helminth infection MSC in mesenteric AT (mAT) become enriched in non-differentiated progenitor cells. This is accompanied by MSC-intrinsic metabolic reprogramming supporting increased secretion of extracellular matrix (ECM), IL-33, and TSLP. In parallel, Th2 resident memory (Th2RM) cells populate the mAT and persist after infection is resolved. These cells express Areg, TGFβ and IL-5, and are necessary to promote infection induced changes within mAT, including MSC reprogramming and tissue eosinophilia. In turn, IL-33 and TSLP from MSC facilitate Th2RM activation and maintenance. Our findings link Th2RM cells to mAT remodeling during intestinal infection, underscoring the reciprocal dependence of stroma and resident immune cells for lasting tissue immunity.
Project description:Cellular plasticity confers cancer cells the ability to adapt to micro-environmental changes, a fundamental requirement for tumour progression and metastasis. The epithelial to mesenchymal transition (EMT) is a transcriptional programme associated with increased cell motility and stemness. Beside EMT, the mesenchymal to amoeboid transition (MAT) has been described during tumour progression but, to date, little is known about its transcriptional control and involvement in stemness. The aim of this study is to investigate (i) the transcriptional profile associated with the MAT programme and (ii) to study whether MAT acquisition in melanoma cancer cells correlate with clonogenic potential to promote tumor growth. Our results demonstrate that MAT programme in melanoma is characterised by increased stemness and clonogenic features of cancer cells, thus sustaining tumour progression. Furthermore, these data suggest that stemness is not an exclusive feature of cells undergoing EMT, but more generally is associated with an increase in cellular plasticity of cancer cells.
Project description:Examined the role of maternal voluntary wheel running commencing 10 weeks prior to gestation, and throughout pregnancy, on placental transcriptome in late gestation While exercise (EX) is beneficial during pregnancy for both mother and child, little is known about the mechanisms by which maternal (MAT EX) mediates changes in utero. We hypothesized that effects of MAT EX prior to and during gestation will be evident in transcriptomic signatures in the placenta and will be sexually dimorphic. Six-week-old female C57BL/6 mice were divided into 2 groups; with (exercise, EX; N = 7) or without (sedentary, SED; N = 8) access to voluntary running wheels. EX was provided via 24-hour access to wheels for 10 weeks prior to conception until late pregnancy (18.5 days post coitum). Sex-stratified placenta and fetal livers were collected. Mi-croarray analysis of SED and EX placenta revealed that MAT EX affected gene transcript expression of 283 and 661 transcripts in male and female placenta (±1.4-fold, p < 0.05). Gene-set enrichment and Ingenuity Pathway analyses of male placenta showed that MAT EX led to inhibition of sig-naling pathways, biological functions, and down regulation of transcripts related to lipid and steroid, while MAT EX in female placentas led to activation of pathways, biological functions, and gene expression related to muscle growth, brain, vascular development, and growth factors. Overall, our results suggest that effects of MAT EX on the placenta and presumably on the offspring are influenced by maternal habitus and are sexually dimorphic.