Gene expression profile of Candida albicans in response to murine IL17A
ABSTRACT: Global transcriptional profiling revealed that IL-17A induced artificial nutrient starvation conditions in Candida albicans, resulting in a downregulated target of rapamycin (TOR) signaling pathway and in increased autophagic responses and intracellular cAMP. We used microarray to detail the global programme of gene expression underlying IL17A sensing by Candida albicans at different time points (T0_0h T1_4h, T2_24h) and identified distinct classes of up-regulated and down regulated genes.
Project description:The liver has the unique capacity to regenerate after surgical resection. However, the regulation of liver regeneration is not completely understood. We performed miRNA microarray analyses of liver tissue from Wistar rats at different time points after 70% partial hepatectomy (0, 2, 6, 12, 24, 48 hours, and 5 days) and after sham laparotomy (12, 24, and 48 hours). We demonstrate that miRNA expression patterns changed during liver regeneration and that these changes were most evident during the peak of DNA replication at 24 hours after resection. Expression of 13 miRNAs, including five members of the let-7 family, was significantly reduced 12-48 hours after resection, whereas 3 miRNAs were significantly upregulated (> 25% change). We provide a temporal miRNA expression dataset of the regenerating rat liver, which indicates a primary function for miRNA during the peak of DNA replication. These data will assist further functional studies on the role of miRNAs during liver regeneration. This project analyzed the miRNA expression in 30 different sample types of the organism rattus norvegicus. The miRNA expression was compared at 7 different growth time points after liver resection (0, 2, 6, 12, 24, 48 hours, 5 days) and at 3 different time points after sham laparotomy (12, 24, and 48 hours). Three biological replicates were used per time point.
Project description:Gene expression changes in L. monocytogenes EGDe during lag-phase associated cold acclimation were succesfully defined through expression profiling The gene expression profiles of L. monocytogenes EGD-e cells that underwent cold acclimation during lag phase were analyzed in a genome-wide microarray assay with a total of 2847 genes, were the resulting significantly regulated genes were splitted in two groups (up-and down-regulated)
Project description:MicroRNAs (miRNAs) are a class of endogenous, non-coding RNAs that mediate post-transcriptional gene silencing by inhibiting mRNA translation and promoting mRNA decay. DICER1, an RNAse III endonuclease encoded by Dicer1, is required for processing short 21-22 nucleotide miRNAs from longer double-stranded RNA precursors. Here, we investigate the loss of Dicer1 in mouse postnatal male germ cells to determine how disruptions in the miRNA biogenesis pathway may contribute to infertility. Reduced levels of Dicer1 transcripts and DICER1 were confirmed in germ cell knock-out (GCKO) testes by postnatal day 18 (P18). Compared to wild-type (WT) at 8 weeks, GCKO males had no change in body weight, yet showed significant reductions in testis mass and sperm number. Histology and fertility tests confirmed spermatogenic failure in GCKO males. Array analyses at P18 showed 96% of miRNA genes were down-regulated and 37% of protein-coding genes were differentially expressed in GCKO testes. Interestingly, we observed preferential overexpression of genes on the sex chromosomes in GCKO testes, with more than 80% of the genes overlapping those proposed to undergo meiotic sex chromosome inactivation (MSCI) in the germ cells. Compared to WT, GCKO mice showed higher percentages of cells at early meiotic stages (leptotene and zygotene) but lower percentages at later stages (pachytene, diplotene and metaphase I), providing evidence that deletion of Dicer1 leads to disruptions in meiotic progression. Furthermore, we observed fewer elongating spermatids with proper translational activation of transition protein 2 (Tnp2), protamine 1 and 2 (Prm1 and Prm2) in GCKO testes after step 12-14. Therefore, deleting Dicer1 in early postnatal germ cells causes misregulation of transcripts encoded by genes on the sex chromosomes, impairs meiotic progression and post-meiotic translational control and results in spermatogenic failure and infertility. Total RNA, including miRNAs, were purified from a total of six individual mouse samples. The tissue collected was obtained from wild-type (control; n=3) and Dicer1 germ cell knockout (mutant; n=3) testes on P18. One miRNA GCKO sample, M36, was determined to be of poor quality and was excluded from the study; thus, a total of five miRNA samples were analyzed.
Project description:We investigate and define here the in vivo biological effects of shikonin, at the transcriptome and microRNA levels, in mouse skin tissues. Through cross-examination between total RNA transcripts and microRNA data sets, we predicted that shikonin treatment may affect the epithelial–mesenchymal transition (EMT) process and the expression of related microRNAs, including 200a, 200b, 200c, 141, 205 and 429 microRNAs, in test skin tissue. Indeed, further in vivo tests on mouse abdominal skin tissue confirmed the stimulatory effect of shikonin on regulatory molecules of the EMT process in epidermal tissue. In addition, RT-PCR analyses confirmed the downregulating effects of shikonin on the expression of microRNA 205 and members of the microRNA 200 family, which are known to be involved in the EMT process. Gene expression of two RNA targets of the microRNA 200 family in EMT regulation, namely Sip1 (Zeb2) and Tcf8 (Zeb1), were consistently upregulated by shikonin treatment. We demonstrate here that shikonin can confer a potent stimulatory effect on the EMT process and suppress the associated-microRNAs expression in vivo in skin tissues. In order to systematically evaluate the effects of shikonin on mouse skin tissues, we next compared the gene expression profiles in shikonin-treated and acetone-treated skin tissues at different time points. Total RNA samples were collected at indicated time points for transcriptome and microRNA array analyses.
Project description:Perforin is a major effector molecule of human natural killer (NK) cells. It can induce delayed growth of Candida albicans hyphae. Here, the fungal transcriptome was analyzed after a co-incubation with 500ng/µl Perforin.
Project description:Aspergillus fumigatus is a ubiquitous mould but also an opportunistic human pathogen causing life-threatening infections in immunocompromised patients. Survival of the fungus in different habitats depends on effective mechanisms of signal perception and transduction such as the cAMP dependent protein kinase A (PKA) pathway, which is involved in virulence of A. fumigatus. Here, by transcriptome analysis putative targets of this important signaling cascade were identified, revealing 632 differently regulated genes including 23 putative transcriptional regulators. The highest up-regulated transcription factor gene was located in the until now unknown fmp secondary metabolite gene cluster encoding an incomplete non-ribosomal peptide synthetase as core enzyme. Overexpression of the fmp cluster resulted in formation of fumipyrrole, which was not described as natural product yet. Although genes of the fmp cluster are transcribed in infected mouse lungs, deletion of its regulatory gene fmpR resulted in wild-type virulence in a murine infection model.
Project description:Invasion of host tissue by the human fungal pathogen, Candida albicans is an important step during many forms of candidosis. However, not all C. albicans strains possess the same invasive and virulence properties. It is known for example that the two clinical isolates SC5314 and ATCC10231 differ in their ability to invade into host tissue and to cause infections. Strain SC5314 is invasive whereas strain ATCC10231 is non-invasive and strongly attenuated in virulence as compared to SC5314. In this study we compare the in vitro transcriptional profiles and the genotypic profiles of these two widely used laboratory strains in order to determine the principal biological and genetic properties which may govern the different potential for invasiveness and virulence. Keywords: transcriptional profiling, comparative genomic hybridisation, invasive vs. non-invasive C. albicans strain Genomic DNA from C. albicans strains SC5314 and ATCC10231 hybridisations were done in duplicate including one dye swap. Total RNA from C. albicans strains SC5314 and ATCC10231 strains were compared in triplicate including one biological replicate and one dye swap.
Project description:Using oligonucleotide microarray analysis, we identified 56 genes that were transcriptionally up-regulated and 69 that were suppressed upon exposure of endothelial cells to C. albicans. Among the regulated genes those attributed to the categories ‘chemotaxis’, ‘signaling’, and ‘transcription and translation’ were remarkably overrepresented. Experiment Overall Design: We performed 4 independent experiments comparing the expression profiles of untreated HUVEC monolayer with those of candida-infected HUVEC monolayer.
Project description:We performed a gene expression analysis of C. albicans SC5314 planktonic cells exposed to the antifungal peptide ApoEdpL-W. Exponentially-growing C. albicans SC5314 cells in SD at 30°C medium were exposed to 2.5 µM ApoEdpL-W and samples were collected after 10 and 30 min. for transcript profiling
Project description:Single-celled organisms have different strategies to sense and utilize nutrients in their ever-changing environments. The opportunistic fungal pathogen Candida albicans is a common member of the human microbiota, especially that of the gastrointestinal (GI) tract. An important question concerns how C. albicans gained a competitive advantage over other microbes to become a successful commensal and opportunistic pathogen. Here, we report that C. albicans uses N-acetylglucosamine (GlcNAc), an abundant carbon source present in the GI tract, as a signal for nutrient availability. When placed in water, C. albicans cells normally enter the G0 phase and remain viable for weeks. However, they quickly lose viability when cultured in water containing only GlcNAc. We term this phenomenon GlcNAc-induced cell death (GICD). GlcNAc triggers the upregulation of ribosomalbiogenesis genes, alterations of mitochondrial metabolism, and the accumulation of reactive oxygen species (ROS), followed by rapid cell death via both apoptotic and necrotic mechanisms. Multiple pathways, including the conserved cyclic AMP (cAMP) signaling and GlcNAc catabolic pathways, are involved in GICD. GlcNAc acts as a signaling molecule to regulate multiple cellular programs in a coordinated manner and therefore maximizes the efficiency of nutrient use. This adaptive behavior allows C. albicans’ more efficient colonization of the gut. Expression profiles of Candida alibcans in three different 5 hours) were determined by high throughput sequencing technology.