Project description:The experiment is to demonstrate the global m6A methylation signatures and their correlation with mRNA expression patterns in microglia stimulated by LPS. The results also identified m6A modifiers with significantly altered expression, which reveals the key one that regulate the m6A modifications in microglia.
Project description:Microglia, the resident immune cells of the brain, can exhibit a broad range of activation phenotypes, many of which have been implicated in several diseases and disorders of the central nervous system including alcohol use disorders and disorders. By utilizing a method optimized for sensitive and rapid quantitative proteomic analysis of microglia involving suspension trapping (S-Trap), we were able to produce efficient and reproducible protein extraction from low cell yielding primary mouse brains. Using a ~2 h gradient on a 75 cm UPLC column with a modified data dependent acquisition method on a hybrid quadrupole-Orbitrap mass spectrometer (QE Plus), 5,062 total proteins were identified where 4,928 of those proteins were quantifiable by label-free quantitation (with 5 biological replicates). This analysis resulted in the most comprehensive proteomic dataset for ethanol- and LPS-treated primary mouse microglia to date and even expanded upon the well-characterized macrophage/microglia response to LPS treatment. This study also highlights the subtle, yet significant changes ethanol exposure can induce when compared to control. Interestingly, these changes are not consistent with the robust classical activation induced by LPS treatment, but instead align with the emerging theory that ethanol-treated microglia yield an alternative activation response. The contrast to LPS-treated microglia leads us to conclude that ethanol does not elicit a strong inflammatory response but rather might have a general inhibitory effect on multiple pathways such as phagocytosis and cell migration.
Project description:Our study demonstrated that the expression of Igf2bp1 in activated microglia was significantly up-regulated, implying a role of Igf2bp1 in LPS-induced m6A modifications in microglia. To understand the roles of Igf2bp1 on LPS-induced m6A modification in microglia, we performed Igf2bp1 loss-of-function (LOF) approach. Microglia stimulated by LPS were transfected with either scrambled siRNA control or Igf2bp1 siRNA for 48 hours. To m6A modification profiles in control and Igf2bp1 LOF microglia were determined by MeRIP-seq analysis.
Project description:We investigated the expression patterns of transcripts in mouse primary microglia that are stimulated by LPS and transfected with miR-106b mimics. Three experiment groups were utilized in the RNA-seq, that are PBS+mimics control group, LPS+mimics group, and LPS+miR-106b mimics group. Samples were collected 2 days after LPS treatment and mimics transfection.
Project description:The role of IL-5 in brain neuroimmunity remains unknown. We investigated the role of IL-5 in cognitive function in aged mice. We examined the transcriptomes of sorted microglia and brain T cells from aged mice treated with PBS or IL-5.
Project description:Primary microglia were derived from neonatal mice and were activated via LPS+ATP treatment. test mice were treated with the drug Ladostigil and transcriptome was compared to untreated controls
Project description:RNA seq of microglia from CPEB1-deficient and LPS-treated mice establish that RNA levels, splicing, and 3' poly(A) site selection are under complex regulation that mediate inflammation and phagaocytosis.
Project description:Curcumin is a potent modulator of the inflammatory transcriptome in microglia We have performed global gene expression analysis of BV-2 microglial cells under the following conditions: untreated, 20 µM Curcumin-treated, 100ng/ml LPS-treated, or 20 µM Curcumin-treated + 100ng/ml LPS-treated