biostudies-arrayexpressJason UlrichMus musculushttps://www.ebi.ac.uk/biostudies/studies/E-MTAB-16169Brain myeloid cells accumulate neutral lipids in multiple human neurodegenerative disorders and relevant mouse models. These neutral lipids are often assumed to be contained in lipid droplets (LDs), and ‘LD-high microglia’ have generally been characterized as maladaptive. While a number of studies have been performed in cell culture and Drosophila models to characterize glial LD dynamics, it is still unclear what roles microglial LD biogenesis play in mammalian tauopathy. To address this question, we induced the deletion of diacylglycerol acyltransferases 1 and 2 (DGATs), enzymes critical for LD formation via triglyceride (TAG) synthesis, from microglia in the PS19 mouse model of tauopathy. We observed that microglial DGAT double KO exacerbated neurodegeneration, induced behavioral deficits, and increased the abundance of brain cholesteryl esters in male PS19 mice. Myeloid cell lipid accumulations appeared to largely localize to endosomes/lysosomes not LDs in PS19 mice at baseline and this phenotype was exacerbated upon DGAT KO. Our results suggest that microglial DGAT-dependent TAG/LD biogenesis is adaptive in advanced tauopathy. Furthermore, the bulk of the lipid accumulation in brain myeloid cells may not correspond to true LDs in this tauopathy model, which has important implications for the development of lipid-modulating therapies for neurodegenerative diseases.biostudies-arrayexpressSample Collection - Frozen cortex from 3 mice of the same genotype were pooled as a single sample. Samples were selected based on those being closest to the mean values of hippocampal volumes. Tissue was homogenized using a Dounce homogenizer in 1 mL of lysis buffer (10 mM Tris-HCl, pH = 7.4; 10 mM NaCl, 3 mM MgCl2; 0.005% NP40; and 0.2 U/μl RNase Inhibitor in nuclease-free water at 4°C) and incubated on ice for 15 min. A 30 μm MACS SmartStrainer was used to remove cell debris and large clumps, followed by centrifugation at 500 x g for 5 min at 4°C. After carefully removing the supernatant, the nuclei pellet was resuspended with 5 mL Nuclei wash and resuspension buffer (1% BSA and 0.2 U/μl RNase Inhibitor in 1 X PBS). Then cell debris removal step, centrifugation, and resuspension were repeated 2 times. Only 500 μl of Nuclei wash and resuspension buffer was added into the last resuspension step. This solution was mixed with 900 μl of Sucrose Cushion Buffer I (2.7 mL Nuclei Pure 2M Sucrose Cushion Solution with 300 μl Nuclei Pure Sucrose Cushion Solution and then carefully layered to the top of 500 μl Sucrose Cushion Buffer I in a 2 mL Eppendorf tube. This sucrose gradient was centrifuged at 13,000 x g for 45 min at 4°C. After centrifugation, the nuclear pellet was resuspended by 500 μl Nuclei Wash and Resuspension Buffer. The nuclei concentration was determined using a Countess with pre-DAPI stain. Finally, nuclei concentration was adjusted to ∼1200 nuclei/μl using Nuclei Wash and Resuspension Buffer followed by proceeding to the 10 x Genomics protocol.Sequencing - The libraries were sequenced at the Genome Technology Access Center (GTAC) using an Illumina Novaseq6000 (Illumina).Sample Treatment - Tamoxifen (200 mg/kg) or corn oil were administered at 3 months of age via oral gavage once per day for 5 consecutive days.Nucleic Acid Extraction - Isolated nulcei were subjected to droplet-based 3′ end massively parallel single-cell RNA sequencing using Chromium Single Cell 3′ Reagent Kits (10x Genomics) following the manufacturer’s instructions.Library Construction - Isolated nulcei were subjected to droplet-based 3′ end massively parallel single-cell RNA sequencing using Chromium Single Cell 3′ Reagent Kits (10x Genomics) following the manufacturer’s instructions.OrganizationMINSEQE ScoreAssays and DataProcessed DataMAGE-TAB FilesData Transformation - Cellranger reads were analyzed using SoupX and Seurat.Sequence Alignment - Reads were aligned to pre mRNA library (mm10) using CellRangerMetabolomicsUnknownTranscriptomicsGenomicsProteomicsIllumina NovaSeq 6000single nucleus RNA sequencingMus musculusJason UlrichfalseInducible deletion of DGAT1 and 2 from microglia exacerbates neurodegeneration and endolysosomal lipid accumulation in male PS19 miceBrain myeloid cells accumulate neutral lipids in multiple human neurodegenerative disorders and relevant mouse models. These neutral lipids are often assumed to be contained in lipid droplets (LDs), and ‘LD-high microglia’ have generally been characterized as maladaptive. While a number of studies have been performed in cell culture and Drosophila models to characterize glial LD dynamics, it is still unclear what roles microglial LD biogenesis play in mammalian tauopathy. To address this question, we induced the deletion of diacylglycerol acyltransferases 1 and 2 (DGATs), enzymes critical for LD formation via triglyceride (TAG) synthesis, from microglia in the PS19 mouse model of tauopathy. We observed that microglial DGAT double KO exacerbated neurodegeneration, induced behavioral deficits, and increased the abundance of brain cholesteryl esters in male PS19 mice. Myeloid cell lipid accumulations appeared to largely localize to endosomes/lysosomes not LDs in PS19 mice at baseline and this phenotype was exacerbated upon DGAT KO. Our results suggest that microglial DGAT-dependent TAG/LD biogenesis is adaptive in advanced tauopathy. Furthermore, the bulk of the lipid accumulation in brain myeloid cells may not correspond to true LDs in this tauopathy model, which has important implications for the development of lipid-modulating therapies for neurodegenerative diseases.2025-11-17T00:00:00Z2026-05-27T12:46:40.927Z2025-11-17T16:27:35.705ZE-MTAB-16169ERP185185EFO_0002944EFO_0004170EFO_0004917EFO_0009809EFO_0005518EFO_0003816EFO_0004184EFO_0003969