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Autofluorescence positive and negative microglia constitute novel subsets found in healthy brain.

ABSTRACT: Utilizing autofluorescence (AF) as a discriminating parameter, we identified two novel microglia subsets in both mice, termed autofluorescence-positive (AF+) and negative (AF-), and characterized them by label-free proteomics.

INSTRUMENT(S): Orbitrap Fusion Lumos

ORGANISM(S): Mus Musculus (mouse)

SUBMITTER: Dirk Walther

LAB HEAD: Dirk Walther

PROVIDER: PXD017505 | Pride | 2020-06-26

REPOSITORIES: Pride

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			Action	DRS
	OL1_191219_DW1_S_MicrogliaPopulations_AFNegative-1.raw	Raw
	OL1_191219_DW1_S_MicrogliaPopulations_AFNegative-2.raw	Raw
	OL1_191219_DW1_S_MicrogliaPopulations_AFNegative-3.raw	Raw
	OL1_191219_DW1_S_MicrogliaPopulations_AFNegative-4.raw	Raw
	OL1_191219_DW1_S_MicrogliaPopulations_AFNegative-5.raw	Raw

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Publications

Differential accumulation of storage bodies with aging defines discrete subsets of microglia in the healthy brain.

Burns Jeremy Carlos JC Cotleur Bunny B Walther Dirk M DM Bajrami Bekim B Rubino Stephen J SJ Wei Ru R Franchimont Nathalie N Cotman Susan L SL Ransohoff Richard M RM Mingueneau Michael M

eLife 20200624

To date, microglia subsets in the healthy CNS have not been identified. Utilizing autofluorescence (AF) as a discriminating parameter, we identified two novel microglia subsets in both mice and non-human primates, termed autofluorescence-positive (AF+) and negative (AF-). While their proportion remained constant throughout most adult life, the AF signal linearly and specifically increased in AF+ microglia with age and correlated with a commensurate increase in si ...[more]

PMID: 32579115

Similar Datasets

Differential accumulation of storage bodies with aging defines discrete subsets of microglia in the healthy brain.

Project description:To date, microglia subsets in the healthy CNS have not been identified. Utilizing autofluorescence (AF) as a discriminating parameter, we identified two novel microglia subsets in both mice and non-human primates, termed autofluorescence-positive (AF+) and negative (AF-). While their proportion remained constant throughout most adult life, the AF signal linearly and specifically increased in AF+ microglia with age and correlated with a commensurate increase in size and complexity of lysosomal storage bodies, as detected by transmission electron microscopy and LAMP1 levels. Post-depletion repopulation kinetics revealed AF- cells as likely precursors of AF+ microglia. At the molecular level, the proteome of AF+ microglia showed overrepresentation of endolysosomal, autophagic, catabolic, and mTOR-related proteins. Mimicking the effect of advanced aging, genetic disruption of lysosomal function accelerated the accumulation of storage bodies in AF+ cells and led to impaired microglia physiology and cell death, suggestive of a mechanistic convergence between aging and lysosomal storage disorders.

| S-EPMC7367682 | biostudies-literature

Microglia constitute a barrier that prevents neurotoxic protofibrillar Aβ42 hotspots around plaques.

Project description:In Alzheimer's disease (AD), β-amyloid (Aβ) plaques are tightly enveloped by microglia processes, but the significance of this phenomenon is unknown. Here we show that microglia constitute a barrier with profound impact on plaque composition and toxicity. Using high-resolution confocal and in vivo two-photon imaging in AD mouse models, we demonstrate that this barrier prevents outward plaque expansion and leads to compact plaque microregions with low Aβ42 affinity. Areas uncovered by microglia are less compact but have high Aβ42 affinity, leading to the formation of protofibrillar Aβ42 hotspots that are associated with more severe axonal dystrophy. In ageing, microglia coverage is reduced leading to enlarged protofibrillar Aβ42 hotspots and more severe neuritic dystrophy. CX3CR1 gene deletion or anti-Aβ immunotherapy causes expansion of microglia coverage and reduced neuritic dystrophy. Failure of the microglia barrier and the accumulation of neurotoxic protofibrillar Aβ hotspots may constitute novel therapeutic and clinical imaging targets for AD.

| S-EPMC4311408 | biostudies-literature

GAP_Culture_conditions

Project description:Gene expression analysis of A. parasiticus grown under conditions conducive and nonconductive for aflatoxin production was evaluated using glass slide microarrays containing 765 ESTs. Aflatoxins (AF) produced by Aspergillus flavus and A. parasiticus are responsible for approximately $270M in losses annually in the corn, peanut, cotton, and tree nut industries of the United States. In an attempt to develop effective control procedures, research has focused on elucidating the pathway of aflatoxin biosynthesis with the goal of identifying potential targets for intervention. Research to date points to the involvement of complex and likely overlapping regulatory circuits. To better understand the regulation by nutritional and environmental factors, a targeted cDNA microarray consisting of genes associated with AF production was employed to investigate the influence of nitrogen source, carbon source, temperature, and pH on AF production. For each nutritional or environmental parameter examined, a condition conducive for aflatoxin production was compared with a condition that was non-conducive. Three replicate cultures for each condition were grown. Cultures were grown for 24 hours and harvested. AF production for each culture was quantified and tissues were stored separately at -80C. For array hybridization, tissue samples for each condition were pooled and RNA extracted using Trizol reagent. Keywords = aspergillus Keywords = aflatoxin Keywords = regulation Keywords = secondary metabolism

2010-06-10 | E-GEOD-1649 | biostudies-arrayexpress

Wild Asparagus Shoots Constitute a Healthy Source of Bioactive Compounds.

Project description:Wild Asparagus shoots are consumed worldwide, although most species remain understudied. In this work, a total of four wild Asparagus species were collected from different locations and analyzed compared with farmed A. officinalis. Shoots were screened for (i) phenolic compounds by HPLC-DAD and LC-MS; (ii) total phenolic acids and total flavonoid content by the Folin-Ciocalteu and aluminum chloride methods; (iii) vitamin C by HPLC-DAD; (iv) antioxidant activity by the DPPH and ABTS•+ methods; and (v) the in vitro antiproliferative activities against HT-29 colorectal cancer cells by the MTT assay. Phenolics ranged from 107.5 (A. aphyllus) to 605.4 mg/100 g dry weight (dw) (A. horridus). Vitamin C ranged from 15.8 (A. acutifolius) to 22.7 mg/100 g fresh weight (fw) (A. officinalis). The antioxidant activity was similar in all species, standing out in A. officinalis with 5.94 (DPPH) and 4.64 (ABTS) mmol TE/100 g dw. Among phenolics, rutin reached the highest values (574 mg/100 g dw in A. officinalis), followed by quercetin, nicotiflorin, asterin, and narcissin. The MTT assay revealed the inhibitory effects of ethanol extracts against HT-29 cancer cells, highlighting the cell growth inhibition exercised by A. albus (300 µg/mL after 72 h exposure to cells). This work improves knowledge on the phytochemicals and bioactivities of the shoots of wild Asparagus species and confirms their suitability for use as functional foods.

| S-EPMC10421306 | biostudies-literature

Arecaceae Seeds Constitute a Healthy Source of Fatty Acids and Phenolic Compounds.

Project description:Seeds of most Arecaceae species are an underutilized raw material that can constitute a source of nutritionally relevant compounds. In this work, seeds of 24 Arecaceae taxa were analyzed for fatty acids (FAs) by GC-FID, for phenolics by HPLC-DAD and LC-MS, and for their antitumor activity against the HT-29 colorectal cancer cell line by the MTT assay. Lauric, oleic, and linoleic acids were the prominent FAs. Cocoseae species contained total FAs at 28.0-68.3 g/100 g seeds, and in other species total FAs were from 1.2 (Livistona saribus) to 9.9 g/100 g (Washingtonia robusta). Sabal domingensis, Chamaerops humilis, and Phoenix dactylifera var. Medjool had unsaturated/saturated FA ratios of 1.65, 1.33-1.78, and 1.31, respectively, and contained 7.4, 5.5-6.3, and 6.4 g FAs/100 g seeds, respectively. Thus, they could be used as raw materials for healthy oilseed production. Phenolics ranged between 39 (Livistona fulva) and 246 mg/100 g (Sabal palmetto), and of these, caffeic acid, catechin, dactylifric acid, and rutin had the highest values. (-)-Epicatechin was identified in most seed extracts by LC-MS. Hydroalcoholic extracts from five species showed a dose-dependent inhibitory effect on HT-20 cells growth at 72 h (GI50 at 1533-1968 µg/mL). Overall, Arecaceae seeds could be considered as a cheap source of health-promoting compounds.

| S-EPMC9867020 | biostudies-literature

CCR2 deficiency alters activation of microglia subsets in traumatic brain injury.

Project description:In traumatic brain injury (TBI), a diversity of brain resident and peripherally derived myeloid cells have the potential to worsen damage and/or to assist in healing. We define the heterogeneity of microglia and macrophage phenotypes during TBI in wild-type (WT) mice and Ccr2-/- mice, which lack macrophage influx following TBI and are resistant to brain damage. We use unbiased single-cell RNA sequencing methods to uncover 25 microglia, monocyte/macrophage, and dendritic cell subsets in acute TBI and normal brains. We find alterations in transcriptional profiles of microglia subsets in Ccr2-/- TBI mice compared to WT TBI mice indicating that infiltrating monocytes/macrophages influence microglia activation to promote a type I IFN response. Preclinical pharmacological blockade of hCCR2 after injury reduces expression of IFN-responsive gene, Irf7, and improves outcomes. These data extend our understanding of myeloid cell diversity and crosstalk in brain trauma and identify therapeutic targets in myeloid subsets.

| S-EPMC8594931 | biostudies-literature

GAP_Culture_conditions

2005-01-01 | GSE1649 | GEO

In situ Patch-seq analysis of microglia reveals a lack of stress genes as found in FACS-isolated microglia.

Project description:We applied the patch-seq technique to harvest transcripts from individual microglial cells from cortex, hippocampus and corpus callosum of acute brain slices from adult mice. After recording membrane currents with the patch-clamp technique, the cytoplasm was collected via the pipette and underwent adapted SMART-seq2 preparation with subsequent sequencing. On average, 4138 genes were detected in 113 cells from hippocampus, corpus callosum and cortex, including microglia markers such as Tmem119, P2ry12 and Siglec-H. Comparing our dataset to previously published single cell mRNA sequencing data from FACS-isolated microglia indicated that two clusters of cells were absent in our patch-seq dataset. Pathway analysis of marker genes in FACS-specific clusters revealed association with microglial activation and stress response. This indicates that under normal conditions microglia in situ lack transcripts associated with a stress-response, and that the microglia-isolation procedure by mechanical dissociation and FACS triggers the expression of genes related to activation and stress.

| S-EPMC11239014 | biostudies-literature

Variation of B cell subsets with age in healthy Malawians.

Project description:Although a number of previous studies have shown that different lymphocyte subsets, including B cells, vary with age, how different B cell subsets vary with age in Malawian population has not been shown before. We recruited Malawian participants of different ages and analyzed their venous blood samples for different B cell subsets. We found that both percentage and absolute counts of B cells varied with age peaking in the 7 to 12 months age group. Proportion of naïve B cells was highest in neonates and decreased with age whereas the percentage of memory B cells was lowest in neonates and increased with age. When we zeroed in on the age band within which the proportion of B cells was highest, both classical and activated memory B cells increased with age and the naïve followed the opposite trend. These results provide additional knowledge in our understanding of the dynamics of B cell subsets in individuals of a specific ethnicity as they age.

| S-EPMC8270464 | biostudies-literature

Competitive repopulation of an empty microglial niche yields functionally distinct subsets of microglia-like cells.

Project description:Circulating monocytes can compete for virtually any tissue macrophage niche and become long-lived replacements that are phenotypically indistinguishable from their embryonic counterparts. As the factors regulating this process are incompletely understood, we studied niche competition in the brain by depleting microglia with >95% efficiency using Cx3cr1CreER/+R26DTA/+ mice and monitored long-term repopulation. Here we show that the microglial niche is repopulated within weeks by a combination of local proliferation of CX3CR1+F4/80lowClec12a- microglia and infiltration of CX3CR1+F4/80hiClec12a+ macrophages that arise directly from Ly6Chi monocytes. This colonization is independent of blood brain barrier breakdown, paralleled by vascular activation, and regulated by type I interferon. Ly6Chi monocytes upregulate microglia gene expression and adopt microglia DNA methylation signatures, but retain a distinct gene signature from proliferating microglia, displaying altered surface marker expression, phagocytic capacity and cytokine production. Our results demonstrate that monocytes are imprinted by the CNS microenvironment but remain transcriptionally, epigenetically and functionally distinct.

| S-EPMC6242869 | biostudies-literature

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