Project description:We report the transfer of alpha-s-ynuclein aggregates from neurons to microglia and examined transcriptomic changes following protein extraction. By obtaining RNA samples, we performed differential expression analysis and generated gene ontology enrichment and network analysis.
Project description:We report the transfer of alpha-s-ynuclein aggregates from neurons to microglia and examined transcriptomic changes following protein extraction. By obtaining RNA samples, we performed differential expression analysis and generated gene ontology enrichment and network analysis.
Project description:Microglia, brain resident macrophages, require instruction from the central nervous system microenvironment to maintain their identity, morphology, and to regulate inflammatory responses. We investigated the heterogeneity of response of microglia to the presence of neurons and astrocytes by performing single-cell sequencing of microglia in both monoculture, and in coculture with neurons and astrocytes.
Project description:DNA double strand breaks (DSBs) are linked to neurodegeneration and senescence. However, it is not clear how DSB-bearing neurons influence neuroinflammation associated with neurodegeneration. Here, we characterize DSB-bearing neurons from the CK-p25 mouse model of neurodegeneration using single-nucleus, bulk, and spatial transcriptomic techniques. DSB-bearing neurons enter a late-stage DNA damage response marked by NFκB-activated senescent and antiviral immune pathways. In humans, Alzheimer’s disease pathology is significantly associated with immune activation in excitatory neurons. Spatial transcriptomics reveal that regions of CK-p25 brain tissue dense with DSB-bearing neurons harbor signatures of inflammatory microglia, which is ameliorated by NFκB knock-down in neurons. Inhibition of NFκB in DSB-bearing neurons also reduces microglia activation in organotypic mouse brain slice culture. In conclusion, DSBs activate immune pathways in neurons, which in turn adopt a senescence-associated secretory phenotype to elicit microglia activation. These findings highlight a novel role for neurons in the mechanism of disease-associated neuroinflammation.
Project description:We developed a novel FACS-based method to isolate enriched populations of neurons, astrocytes, and microglia. Whole mouse brains were digested, and populations of neurons, astrocytes, and microglia were subsequently enriched by FACS using a combination of positive gating on cell-type specific markers and stringent negative selection against markers of oligodendrocytes and endothelial cells. We confirmed a strong enrichment of corresponding cell-type specific genes in isolated populations and a reduction in gene markers of endothelial cells and oligodendrocytes, demonstrating that highly pure populations of neurons, astrocytes, and microglia were obtained.
Project description:We investigated non-cell-autonomously regulated gene expression in microglia, neurons and astrocytes by co-culturing these cell types (derived from different mammalian species) together, and then separating the RNA-seq reads from each cell type/species in silico.