Project description:IFN-gamma is a classical microglial stimulant. We used microarrays to investigate the microglial gene regulatory network activated by interferon-gamma. Experiment Overall Design: Primary rat microglia cultures were established and maintained for 15 days. For activation studies, fresh media containing IFN-gamma (100 U/ml) were added and left overnight (16 hours). Total RNA was extraction and hybridized on Affymetrix microarrays (RG_U34A). Five arrays were run from total RNA derived from unstimulated 5 independent cultures (Mgl_CON1, Mgl_CON2, Mgl_CON3, Mgl_CON5 and Mgl_CON6). Three standard microglial cultures were stimulated (Mgl_IFNgamma_2 and Mgl_IFNgamma_3) with one of the samples analysed in triplicate as a technical control (Mgl_IFNgamma_1a, Mgl_IFNgamma_1b and Mgl_IFNgamma_1c).
Project description:In order to establish a rat embryonic stem cell transcriptome, mRNA from rESC cell line DAc8, the first male germline competent rat ESC line to be described and the first to be used to generate a knockout rat model was characterized using RNA sequencing (RNA-seq) analysis.
Project description:Microglia become activated by disturbances in the homeostasis of their local microenvironment. While there are varying degrees of microglia activation, a pro-inflammatory reactive state induced by exposure to stimuli like LPS and IFN-γ. In this study, we identified a total of 5497 proteins in whole cell proteome and 4938 proteins for secretome of activated BV-2 mouse microglia cell line with LPS or IFN-γ using improved shotgun proteomic approach. From the differentially expressed proteins in stimulated microglia, we were able to classify pathways related immune-inflammatory response or metabolism. Moreover, we performed longitudinal quantification of secreted proteins during the detrimental activation of microglia which releases neurotoxic molecules mediated neuronal cell loss in the brain region.
