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ScRNA-seq of microglia from Cx3cr1CreERT2(Litt) adult mice reveals gene expression profile after TAM treatment.

ABSTRACT: scRNA-seq of microglia from Cx3cr1CreERT2(Litt) adult mice reveals gene expression profile after TAM treatment.

PROVIDER: PRJNA1302488 | ENA |

REPOSITORIES: ENA

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scRNA-seq of microglia from Cx3cr1CreERT2(Litt) adult mice reveals gene expression profile after TAM treatment.

Project description:The Cx3cr1CreERT2(Litt) mouse model is widely used for in-depth studies of myeloid cells, including microglia. In this study, we employed single-cell RNA sequencing (scRNA-seq) to characterize microglial responses to tamoxifen (TAM) administration in adult Cx3cr1CreERT2(Litt) mice. Our analysis revealed a prolonged TAM-dependent induction of CDKN1A in the microglial population, highlighting a effect of the Cre-induction system itself.

2025-08-15 | GSE304771 | GEO

Effect of TAM treatment on gene expression profile of microglia from Cx3cr1CreERT2 mouse lines.

Project description:Microglia, the resident macrophages in the central nervous system (CNS), have been intensively studied using rodent genetic models, including the Cx3cr1-Cre mouse line. These mice have enabled the in-depth analyses of the biological features and functions of myeloid cells, including microglia. Occasionally, these Cx3cr1CreERT2 tools have yielded conflicting outcomes, the underlying mechanism of which remains unclear. Here, we comparatively characterize the two available Cx3cr1CreERT2 lines (Cx3cr1CreERT2(Litt) and Cx3cr1CreERT2(Jung)). We find a mouse line-specific and TAM-dependent persistent induction of CDKN1A in microglia of Cx3cr1CreERT2(Litt) mice, but not in those of Cx3cr1CreERT2(Jung) mice, which affects experimental readouts with altered proliferative capacity. Furthermore, aberrant cellular alterations observed in postnatal Cx3cr1CreERT2(Litt) microglia are mitigated by a functional inhibition of CDKN1A. Together, these findings underscore the significance of mouse line-specific phenomena that alter microglial outcomes in a CDKN1A-dependent manner, highlighting the importance of rigorous validation of genetic mouse tools.

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Microglia use TAM receptors to detect and engulf amyloid beta plaques

Project description:Two microglial TAM receptor tyrosine kinases - Axl and Mer - have been linked to Alzheimer’s disease, but their roles in disease have not been tested experimentally. We find that in Alzheimer’s disease and its mouse models, induced expression of Axl and Mer in amyloid plaque-associated microglia is coupled to induced plaque decoration by the TAM ligand Gas6 and its co-ligand phosphatidylserine. In the APP/PS1 mouse model of Alzheimer’s disease, sIngle cell RNAseq analysis comparing wild type microglia with those with Axl and Mer deficiency reveals a similar disease state transitional program of microglia but a dampened differential expression of numerous AD siganture genes in microglia lacking TAM receptors. In line with the transcriptomic data, using two-photon microscopy, we show that genetic ablation of Axl and Mer results in microglia that are unable to normally detect, respond to, organize, or phagocytose amyloid beta plaques. These major deficits notwithstanding, and contrary to expectation, TAM-deficient APP/PS1 mice develop fewer dense-core plaques than APP/PS1 mice with normal microglia. Our findings reveal that the TAM system is an essential mediator of microglial recognition and engulfment of amyloid plaques, and that TAM-driven microglial phagocytosis does not constrain, but rather promotes, plaque development.

2021-02-25 | GSE160523 | GEO

Effect of TAM treatment on gene expression profile of microglia from Cx3cr1CreERT2 mouse lines.

Project description:Effect of TAM treatment on gene expression profile of microglia from Cx3cr1CreERT2 mouse lines.

| PRJNA1302487 | ENA

AffyExon1 genearray expression profile of adult mouse microglia and splenic Ly6C monocytes

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Microglia use TAM receptors to detect and engulf amyloid beta plaques

Project description:Microglia use TAM receptors to detect and engulf amyloid beta plaques

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Proteomics of tumor associated microglia, tumor tissue from SB28 glioblastoma mice, and control microglia

Project description:The 18 kDa translocator protein (TSPO) emerges as an important PET biomarker to assess the tumor microenvironment (TME) in glioblastoma. However, various cellular sources hamper interpretation and biological understanding of TSPO and other immune biomarkers in the TME. Thus, we established a novel method, combining immunomagnetic cell sorting after radiotracer injection (scRadiotracing) with 3D histology via light sheet microscopy and proteomics to dissect cellular allocation of TSPO enrichment in glioblastoma. Single tumor cells of implanted SB28 glioblastoma mice indicated 1.37-fold higher TSPO tracer uptake and 1.46-fold higher TSPO protein expression levels when compared to tumor associated microglia/macrophages (TAMs). Using proteomics, we compared the proteome of tumor associated microglia/macrophages (TAMs), Tumor tissue (TT) and control microglia from WT mice without glioblastoma. This analysis identified TAM specific targets for PET radioligand development with additional potential to monitor diverse TAM subpopulations in vivo. In summary, our data indicate that tumor cells need to be acknowledged as the main contributor to TSPO as a biomarker in glioblastoma. Combining cellular tracer uptake measures with 3D histology facilitates precise allocation of complex PET signal sources and will serve to validate novel TAM specific radioligands.

2023-10-30 | PXD044033 | Pride

scRNA-seq study of hippocampal microglia, perivascular macrophages and perivascular fibroblasts in pre-plaque APP-KI mice

Project description:Aim of this experiment was to characterize whether SPP1 released from the hippocampal perivascular space could imprint microglia transcriptional states. We performed scRNA-seq analysis on sorted CD140a+ perivascular fibroblast (PVF), CD45highCD11intCD206+ perivascular macrophage (PVM) and CD45intCD11intCX3CR1high microglial cell from dissected hippocampi of 6-month old wild type vs SPP1KO/KO vs AppNL-F mice vs AppNL-FxSpp1KO/KO mice. Cells were isolated from dissected hippocampi as previously described (Sala Frigerio et al.,2019).

2024-02-09 | E-MTAB-11918 | biostudies-arrayexpress

In vivo mouse microglia scRNA seq

Project description:In vivo mouse microglia scRNA seq

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Proteome Dataset for Adult-derived Mouse Microglia

Project description:We report methods development for quantitative proteomic analysis of samples with limited number of cells. The improvements include suspension trapping (S-Trap) for sample processing, longer analytical column length (75cm), and a segmented data dependent acquisition approach. The modified approach was applied to the proteomic analysis of adult-derived mouse microglia and optimized to the cell count expected from mouse brain (~300K microglial cells). With the improved approach, over 4500 quantifiable proteins can be obtained (in biological triplicate) with an average CV of 10%.

2019-04-18 | PXD012006 | Pride

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