Flow Cytometry Analysis of Microglial Phenotypes in the Murine Brain During Aging and Disease DOI Creative Commons

Jillian E J Cox,

Kevin D Pham,

Alex W Keck

и другие.

BIO-PROTOCOL, Год журнала: 2024, Номер 14(1347)

Опубликована: Янв. 1, 2024

Microglia, the brain's primary resident immune cell, exists in various phenotypic states depending on intrinsic and extrinsic signaling. Distinguishing between these phenotypes can offer valuable biological insights into neurodevelopmental neurodegenerative processes. Recent advances single-cell transcriptomic profiling have allowed for increased granularity better separation of distinct microglial states. While techniques such as immunofluorescence RNA sequencing (scRNA-seq) are available to differentiate functions, methods present notable limitations, including challenging quantification methods, high cost, advanced analytical techniques. This protocol addresses limitations by presenting an optimized cell preparation procedure that prevents ex vivo activation a flow cytometry panel distinguish four from murine brain tissue. Following preparation, fluorescent antibodies were applied label 1) homeostatic, 2) disease-associated (DAM), 3) interferon response (IRM), 4) lipid-droplet accumulating (LDAM) microglia, based gene markers identified previous scRNA-Seq studies. Stained cells analyzed assess distribution function age sex. A key advantage this is its adaptability, allowing provided be enhanced using additional with appropriate analyzer (i.e., Cytek Aurora 5 laser spectral cytometer) interrogating different regions or disease models. Additionally, does not require sorting, resulting relatively quick straightforward experiment. Ultimately, compare experimental groups, state age, lower cost higher throughput than scRNA-seq. Key features • Analysis without need imaging, (LDAM), (IRM) microglia any region and/or model interest. modified incorporate interest dyes when capable multiple color detections.

Язык: Английский

Sex-specific influences of APOEε4 genotype on hippocampal neurogenesis and progenitor cells in middle-aged rats DOI Creative Commons
Bonnie H. Lee,

Melike Cevizci,

Stephanie E. Lieblich

и другие.

Biology of Sex Differences, Год журнала: 2025, Номер 16(1)

Опубликована: Фев. 5, 2025

Abstract Background Alzheimer’s disease (AD) disproportionately and uniquely affects females, these sex differences are further exacerbated by the presence of Apolipoprotein (APOE) ε4 alleles, top genetic risk factor for late-onset AD. To expand our understanding about how AD might differentially influence males this study explores APOEε4 hippocampal neurogenesis microglia, key neuroplastic markers involved in pathogenesis, differently middle-aged rats. Methods A rat model expressing humanized (h) allele was characterized to examine adult (neural progenitor cells new-born neurons) immune (microglia) dentate gyrus hippocampus 13 month-old male female Results We observed basal at middle age, as wildtype rats had greater densities neural neurons than Male hAPOEε4 exhibited fewer cells, neurons, more microglia On other hand, Interestingly, females regardless genotype. Correlations were conducted elucidate any relationships between biomarkers. Notably, there a significant positive correlation negative but only Conclusion In contrast clear pattern effects on males, unaltered levels increased density neurons. Furthermore, significantly correlated within not females. This suggests that may be presenting compensatory response genotype age. Collectively, results exemplify importance thoroughly examining influences endophenotypes, it reveal sex-specific pathways protective mechanisms relevant

Язык: Английский

Процитировано

0
Effects of Pesticide Exposure on Neuroinflammation and Microglial Gene Expression: Relevance to Mechanisms of Alzheimer's Disease Risk DOI Open Access
Isha Mhatre‐Winters,

Aseel Eid,

Nicole Blum

и другие.

bioRxiv (Cold Spring Harbor Laboratory), Год журнала: 2025, Номер unknown

Опубликована: Фев. 20, 2025

Abstract Background Alzheimer’s disease (AD) is characterized by the presence of amyloid-β plaques, neurofibrillary tangles, and neuroinflammation. Previously, we reported serum levels dichlorodiphenyldichloroethylene (DDE), primary metabolite pesticide dichlorodiphenyltrichloroethane (DDT), were significantly higher in AD patients compared to age-matched controls that DDT exposure worsened pathology animal models. Objective Here, investigated effect on neuroinflammation mouse microglia (PMG) C57BL/6J mice. Methods Effects inflammation disease-associated determined Results PMG exposed (0.5-5.0 µM) elicited a ∼2-3-fold increase Il-1b mRNA levels, with similar concentration-dependent upregulation Il-6, Nos2, Tnfa . These effects blocked sodium channel antagonist tetrodotoxin, demonstrating role DDT-microglial interactions mediating this response. Additionally, NOS2 protein increased ∼1.5-2-fold, while TNFa was elevated 2-4-fold. male female mice (30 mg/kg) demonstrated Nos2 , Il-6 frontal cortex (1.5-2.3-fold), Il-1b, (1.5-1.8-fold) hippocampus. Furthermore, microglial homeostatic genes, Cx3cr1 P2ry12, Tmem119 downregulated, stage 1 genes upregulated both vitro vivo Notably, Apoe Trem2 only hippocampus females. Conclusion data indicate increases neuroinflammation, which may result from direct actions microglia, providing novel pathway contribute risk.

Язык: Английский

Процитировано

0
Sex chromosomes and gonads modify microglial-mediated pathology in a mouse model of Alzheimer’s disease DOI Creative Commons
Brad T. Casali,

Li Lin,

Olesia Benedict

и другие.

Journal of Neuroinflammation, Год журнала: 2025, Номер 22(1)

Опубликована: Март 13, 2025

Alzheimer's disease (AD) is a neurodegenerative disorder disproportionally affecting women with sex-specific manifestations and therapeutic responses. Microglial-mediated inflammation occurs in response to perpetuates processes, fundamental sex differences microglia may contribute these biases. Both chromosomes gonad-derived hormones shape immune responses, but their contribution immune-mediated mechanisms underlying the bias AD unclear. Crossing Four Core Genotype (FCG) model separate chromosome hormone effects 5xFAD model, we found complement impacted microgliosis, neuroinflammation, plaque burden neuritic dystrophy. Modification of pathology largely correlated influenced remodeling microglial CD11c expression. Our results provide potential trajectories for studying targeting microglial-mediated emphasize complex interplay between during AD.

Язык: Английский

Процитировано

0
Do microglia metabolize fructose in Alzheimer’s disease? DOI Creative Commons

Annalise M. Sturno,

James E. Hassell, Miguel A. Lanaspa

и другие.

Journal of Neuroinflammation, Год журнала: 2025, Номер 22(1)

Опубликована: Март 15, 2025

Alzheimer's disease (AD) is an age-associated neurodegenerative disorder with a complex etiology. While emerging AD therapeutics can slow cognitive decline, they may worsen dementia in certain groups of individuals. Therefore, alternative treatments are much needed. Microglia, the brain resident macrophages, have potential to be novel therapeutic targets as regulate many facets AD, including lipid droplet (LD) accumulation, amyloid beta (Aβ) clearance, and neuroinflammation. To carry out such functions, microglia undergo phenotypic changes, which linked shifts metabolism substrate utilization. homeostatic driven by oxidative phosphorylation (OXPHOS) glycolysis, aging shift further towards glycolysis. Interestingly, this "metabolic reprogramming" increase fructose metabolism. In brain, predominantly express transporter SLC2A5 (GLUT5), enzymes involved fructolysis endogenous production, their expression being upregulated disease. Here, we review evidence for uptake, breakdown, production microglia. We also evaluate literature targeting periphery assess its ability modulate microglial function AD. The transport utilize fructose, coupled well-established role metabolic dysfunction, supports notion that target

Язык: Английский

Процитировано

0
Atg5 in microglia regulates sex-specific effects on postnatal neurogenesis in Alzheimer’s disease DOI Creative Commons

Ellen Walter,

Gabrielle Angst,

Justin L. Bollinger

и другие.

npj Aging, Год журнала: 2025, Номер 11(1)

Опубликована: Март 16, 2025

Abstract Female Alzheimer’s disease (AD) patients display greater cognitive deficits and worse AD pathology as compared to male patients. In this study, we found that conditional knockout (cKO) of Atg5 in female microglia failed obtain disease-associated (DAM) gene signatures familiar mouse model (5xFAD). Next, analyzed the maintenance neurogenesis neural stem cells (NSCs) hippocampus subventricular zone (SVZ) from 5xFAD mice with cKO. Our data indicated cKO reduced NSC number but not mice. However, SVZ, only impaired NSCs Interestingly, 5xFAD; Fip200 Atg14 did show defects. These autophagy genes exhibited a higher activity their SVZ. Together, our indicate sex-specific role for microglial postnatal

Язык: Английский

Процитировано

0
Association of soluble TREM2 with Alzheimer’s disease and mild cognitive impairment: a systematic review and meta-analysis DOI Creative Commons

Ruiqi Wang,

Yijun Zhan, Wenyan Zhu

и другие.

Frontiers in Aging Neuroscience, Год журнала: 2024, Номер 16

Опубликована: Май 22, 2024

Objective Soluble triggering receptor expressed on myeloid cells 2 (sTREM2) is a potential neuroinflammatory biomarker linked to the pathogenesis of Alzheimer’s disease (AD) and mild cognitive impairment (MCI). Previous studies have produced inconsistent results regarding sTREM2 levels in various clinical stages AD. This study aims establish correlation between AD progression through meta-analysis cerebrospinal fluid (CSF) blood. Methods Comprehensive searches were conducted PubMed, Embase, Web Science, Cochrane Library identify observational reporting CSF blood patients, MCI healthy controls. A random effects was used calculate standardized mean difference (SMD) 95% confidence intervals (CIs). Results Thirty-six involving 3,016 3,533 4,510 controls included. significantly higher both [SMD = 0.28, CI (0.15, 0.41)] groups 0.30, (0.13, 0.47)] compared control group. However, no significant differences expression detected 0.09, (−0.09, 0.26)]. Furthermore, increased plasma associated with risk 0.42, (0.01, 0.83)]. Conclusion are positively an MCI. Plasma notably group than may serve as promising for diagnosing not effective distinguishing different Further investigations needed explore longitudinal changes levels, particularly during progression. Systematic review registration https://www.crd.york.ac.uk/prospero/display_record.php?ID=CRD42024514593

Язык: Английский

Процитировано

3
Crosstalk between the DNA damage response and cellular senescence drives aging and age-related diseases DOI
Ajmal Ahmad,

Anneliesse Braden,

Sazzad Khan

и другие.

Seminars in Immunopathology, Год журнала: 2024, Номер 46(3-4)

Опубликована: Авг. 1, 2024

Язык: Английский

Процитировано

3
Minding the Gap: Exploring Neuroinflammatory and Microglial Sex Differences in Alzheimer’s Disease DOI Open Access
Erin G. Reed-Geaghan, Phaedra R. Keller-Norrell

International Journal of Molecular Sciences, Год журнала: 2023, Номер 24(24), С. 17377 - 17377

Опубликована: Дек. 12, 2023

Research into Alzheimer's Disease (AD) describes a link between AD and the resident immune cells of brain, microglia. Further, this suspected is thought to have underlying sex effects, although mechanisms these effects are only just beginning be understood. Many insights result policies put in place by funding agencies such as National Institutes Health (NIH) consider biological variable (SABV) move towards precision medicine due continued lackluster therapeutic options. The purpose review provide an updated assessment current research that summarizes differences pertaining microglia their varied responses AD.

Язык: Английский

Процитировано

8
Sex matters: The MouseX DW-ALLEN Atlas for mice diffusion-weighted MR imaging DOI Creative Commons
Patricia Martínez-Tazo,

Alexandra Santos,

Mohamed Kotb Selim

и другие.

NeuroImage, Год журнала: 2024, Номер 292, С. 120573 - 120573

Опубликована: Март 21, 2024

Overcoming sex bias in preclinical research requires not only including animals of both sexes the experiments, but also developing proper tools to handle such data. Recent work revealed sensitivity diffusion-weighted MRI glia morphological changes response inflammatory stimuli, opening up exciting possibilities characterize inflammation a variety models pathologies, great majority them available mice. However, there are limited resources dedicated mouse imaging, like those required for data processing and analysis. To fill this gap, we build template structural diffusion contrasts, with anatomical annotation according Allen Mouse Brain Atlas, most detailed public resource brain investigation. achieve standardized resource, use large cohort vivo, include sexes. prove utility integrate imaging molecular data, demonstrate significant association between mean diffusivity from gene expression-based density. need equitable representation, compared across warp fields needed match male-based template, our built Then, templates analysing mice obtained different ages, demonstrating that using creates spurious effects, present otherwise. All all, MouseX DW-ALLEN Atlas will be widely useful getting us one step closer healthcare.

Язык: Английский

Процитировано

1
Neuroimaging correlates of Alzheimer's disease biomarker concentrations in a racially diverse high‐risk cohort of middle‐aged adults DOI Creative Commons
Maria Misiura,

Chinkuli Munkombwe,

Kay C. Igwe

и другие.

Alzheimer s & Dementia, Год журнала: 2024, Номер 20(9), С. 5961 - 5972

Опубликована: Авг. 13, 2024

Abstract INTRODUCTION In this study, we investigated biomarkers in a midlife, racially diverse, at‐risk cohort to facilitate early identification and intervention. We examined neuroimaging measures, including resting state functional magnetic resonance imaging (fMRI), white matter hyperintensity vo (WMH), hippocampal volumes, alongside cerebrospinal fluid (CSF) markers. METHODS Our data set included 76 cognitively unimpaired, middle‐aged, Black Americans ( N = 29, F/M 17/12) Non‐Hispanic White 47, 27/20) individuals. compared phosphorylated tau141 amyloid beta (Aβ)42 fMRI default mode network (DMN) subnetwork connectivity, WMH volumes. RESULTS Results revealed significant race × Aβ42 interaction Americans: lower was associated with reduced DMN connectivity increased volumes regions but not non‐Hispanic DISCUSSION findings suggest that precuneus temporal WMHs may be linked Alzheimer's disease risk pathology during middle age, particularly Americans. Highlights Cerebrospinal relates Black, White, Higher volume CSF Precuneus hub for changes detected by connectivity.

Язык: Английский

Процитировано

1