Project description:Since few years, yeast cell wall components and especially beta-glucans (BG) have been identified as potential stimulators of the innate immune system in murine and human species. A large screening of crude and BG-enriched components from Saccharomyces cerevisiae has been performed on murine macrophages. A priming effect with bacterial ligands was revealed in terms of pro-inflammatory cytokines release. To further characterize the responsiveness of bone marrow derived-macrophages which express high levels of dectin-1, the major receptor of BG, transcriptional analysis would provide more hypotheses about the genes and signaling pathways triggered by yeast cell wall compounds in macrophages. Gene expression profiling was performed in murine BMDM pretreated with a set of three BG-containing cell wall compounds and then stimulated with LPS. Conditions with BG-enriched pretreatment clustered together in contrast to the crude compound and mock pretreatment conditions that remained separated whatever the time point analyzed, with a greater number of differentially-expressed genes following the crude compound treatment compared to BG-enriched pretreatment. BG-enriched pretreatment induced a specific set of genes in mouse macrophages, involving the PI3K/AKT signaling pathway. Among them, the two main upregulated genes by BG-enriched priming condition were considered for further analysis and their associated-protein production were consistently increased in BMDM pretreated with BG-enriched compound. Microarray results were confirmed using RT-qPCR on a different set of samples.
Project description:PURPOSE: To provide a detailed gene expression profile of the normal postnatal mouse cornea. METHODS: Serial analysis of gene expression (SAGE) was performed on postnatal day (PN)9 and adult mouse (6 week) total corneas. The expression of selected genes was analyzed by in situ hybridization. RESULTS: A total of 64,272 PN9 and 62,206 adult tags were sequenced. Mouse corneal transcriptomes are composed of at least 19,544 and 18,509 unique mRNAs, respectively. One third of the unique tags were expressed at both stages, whereas a third was identified exclusively in PN9 or adult corneas. Three hundred thirty-four PN9 and 339 adult tags were enriched more than fivefold over other published nonocular libraries. Abundant transcripts were associated with metabolic functions, redox activities, and barrier integrity. Three members of the Ly-6/uPAR family whose functions are unknown in the cornea constitute more than 1% of the total mRNA. Aquaporin 5, epithelial membrane protein and glutathione-S-transferase (GST) omega-1, and GST alpha-4 mRNAs were preferentially expressed in distinct corneal epithelial layers, providing new markers for stratification. More than 200 tags were differentially expressed, of which 25 mediate transcription. CONCLUSIONS: In addition to providing a detailed profile of expressed genes in the PN9 and mature mouse cornea, the present SAGE data demonstrate dynamic changes in gene expression after eye opening and provide new probes for exploring corneal epithelial cell stratification, development, and function and for exploring the intricate relationship between programmed and environmentally induced gene expression in the cornea. Keywords: other