Journal: Nature Communications

Article Title: Microglia-specific regulation of lipid metabolism in Alzheimer’s disease revealed by microglial depletion in 5xFAD Mice

doi: 10.1038/s41467-025-64161-z

Figure Lengend Snippet: A Experimental timeline and interventions. Left: Pharmacological microglial depletion in non-transgenic (Non-Tg) and 5xFAD transgenic mice (mixed B6SJL background) using short-term (2-week) or long-term (3.5 months) treatment. Right: Genetic microglial ablation (FIRE) in 5xFAD mice (B6 background). OSD OpenStandard Diet (Research Diets). Brain samples were collected at indicated time points and analyzed by shotgun lipidomics, NanoString Glial Profiling Panel, and immunofluorescence. Schematic created in BioRender. Xu, Z. (2025) https://BioRender.com/6dlymhx . B –J Pathological characterization after long-term intervention. Representative images and quantifications of microglial marker (P2Y12) ( B –D ), amyloid beta (Aβ, MOAB-2) ( E –G ), and astrocytic marker (GFAP) ( K –M ) immunofluorescence, as well as Methoxy-X04 ( H –J ) fluorescent staining for fibrillar β-sheet amyloid plaques. Each data point represents a brain section, two sections from 3–4 female mice/group were quantified. BZ-X800 Analyzer auto function was used to set a threshold for each section. Representative images shown at matched magnifications and thresholds. N –P APP and neuronal levels. Western blot analysis of APP using 6E10 antibody and synaptic marker Homer1 ( N ), with corresponding quantifications displayed in panels ( O ) and ( P ), respectively. Each data point represents one animal, n = 4–5 mice/group. Trem2 mRNA quantification in both long-term pharmacological ( Q ) and ( R ) genetic cohorts. Each data point represents one animal, n = 4–6 mice/group. All data presented as mean ± SEM. Statistical analysis was performed in GraphPad using one-way ANOVA with Tukey’s post-hoc correction. Representative images (indicated by stars in the corresponding dot plots) were chosen from brain sections present on the same slide, with each slide containing sections from each experimental group. Scale bars: 50 μm for main panels ( B – K ); 20 μm for insets.

Article Snippet: 5xFAD mice (Tg(APPSwFlLon,PSEN1*M146L*L286V)6799Vas/Mmjax), were obtained from the Mutant Mouse Resource and Research Center (MMRRC) at The Jackson Laboratory in two different genetic backgrounds (B6SJL: MMRRC_034840-JAX; and C57BL6: MMRRC_034848-JAX).

Techniques: Transgenic Assay, Immunofluorescence, Marker, Staining, Western Blot

Journal: Nature Communications

Article Title: Microglia-specific regulation of lipid metabolism in Alzheimer’s disease revealed by microglial depletion in 5xFAD Mice

doi: 10.1038/s41467-025-64161-z

Figure Lengend Snippet: A –D Quantification of specific BMP species levels in human postmortem temporal lobe Brodmann area 38 (BA38): comparison of AD vs non-AD controls. Human data normalized by log 10 transformation. Statistical analysis was performed in MetaboAnalyst using metadata table module linear model adjusted for sex and gray/white matter ratio (GWR) with false discovery rate correction for multiple comparisons (FDR = 0.05). E –H Levels of corresponding BMP species in mouse brains following long-term pharmacological microglial depletion. Data normalized by square root transformation and pareto scaling. Statistical analysis was performed in MetaboAnalyst using one-factor module t-testing correcting for multiple comparisons (FDR = 0.05). I –L Levels of same BMP species in mouse brains after genetic microglial depletion. Data normalized by log 10 transformation. Statistical analysis was performed in MetaboAnalyst using metadata table module linear model adjusted for sex correcting for multiple comparisons (FDR = 0.05). Heatmaps of lysosomal gene expression after long-term pharmacological ( M ) and genetic ( N ) microglial depletion. Significantly altered genes denoted: 5xFAD vs Non-Tg (#), and 5xFAD + PLX5622 vs 5xFAD (*) by two-tailed unpaired t-tests with Benjamini-Hochberg correction. Correlations between relative AA-BMP levels and total lysosome gene counts in microglia-depleted mouse brains following long-term pharmacological ( O ) and genetic ( P ) interventions. Statistical analysis was performed in MetaboAnalyst using metadata table module linear model regressing relative AA-BMP levels against all NanoString predefined pathways adjusting for multiple correlations (FDR = 0.05, significance threshold q ≤ 0.05). Each data point represents one animal, n = 4-8 mice/group. All data presented as mean ± SEM.

Article Snippet: 5xFAD mice (Tg(APPSwFlLon,PSEN1*M146L*L286V)6799Vas/Mmjax), were obtained from the Mutant Mouse Resource and Research Center (MMRRC) at The Jackson Laboratory in two different genetic backgrounds (B6SJL: MMRRC_034840-JAX; and C57BL6: MMRRC_034848-JAX).

Techniques: Comparison, Transformation Assay, Gene Expression, Two Tailed Test

Journal: Nature Communications

Article Title: Microglia-specific regulation of lipid metabolism in Alzheimer’s disease revealed by microglial depletion in 5xFAD Mice

doi: 10.1038/s41467-025-64161-z

Figure Lengend Snippet: Relative Grn mRNA expression after long-term pharmacological ( A ) and genetic ( B ) microglial depletion. Correlation analysis of relative AA-BMP levels with relative Grn mRNA expression in long-term pharmacological ( D ) and genetic ( E ) microglial depletion. Relative progranulin protein levels assessed via Western blot ( C ) were correlated with AA-BMP levels ( F ). Each data point represents one animal, n = 4-8 mice/group. Statistical analysis was performed in MetaboAnalyst using metadata table module linear model regressing relative AA-BMP levels against all genes with false discovery rate correction for multiple comparisons (FDR = 0.05). Representative immunofluorescence images showing progranulin (PGRN), Iba1 (microglia/macrophages), MOAB-2 (amyloid plaques) and DRAQ5 (nuclei) in non-Tg ( G ), 5xFAD ( H ) and 5xFAD + PLX5622 ( I ). J Quantification of high progranulin intensity, primarily in plaque-associated and activated microglia. Each data point represents a brain section, 1-2 sections from 3 female mice/group were quantified. All data presented as mean ± SEM, normalized to non-Tg controls. Statistical analysis was performed in GraphPad using ordinary one-way ANOVA with Tukey’s post hoc correction.

Article Snippet: 5xFAD mice (Tg(APPSwFlLon,PSEN1*M146L*L286V)6799Vas/Mmjax), were obtained from the Mutant Mouse Resource and Research Center (MMRRC) at The Jackson Laboratory in two different genetic backgrounds (B6SJL: MMRRC_034840-JAX; and C57BL6: MMRRC_034848-JAX).

Techniques: Expressing, Western Blot, Immunofluorescence

Journal: Alzheimer's & Dementia

Article Title: Degradation of perineuronal nets in hippocampal CA2 explains the loss of social cognition memory in Alzheimer's disease

doi: 10.1002/alz.70813

Figure Lengend Snippet: Progressive disruption of Hippocampal CA2 PNNs in 5XFAD mice. (A) Confocal micrographs showing immunohistochemical expression of WFA (magenta)‐labeled PNNs in coronal half‐brain sections from control and 5XFAD mice. Magnified areas in white rectangles highlight disruption of PNNs around hippocampal CA2 pyramidal neurons labeled with PCP4 (green). Scale 500 µm main images, 50 µm magnified images. (B) High magnification confocal micrographs of a single CA2 pyramidal neuron showing PNN disruption in 5XFAD and consequent loss of characteristic high‐intensity peaks and valley pattern in the line intensity profile (bottom). A line was drawn along the WFA signal in the PCP4 neuron periphery in control and 5XFAD group, showing many high‐intensity WFA peaks in control (dark magenta line) compared to the 5XFAD (light magenta line) group. Scale 2 µm. (C–F) Representative confocal micrographs of PNN (WFA‐magenta) immunofluorescence associated with CA2 pyramidal neurons (PCP4‐green) in 5XFAD mice at 3 (C), 6 (D), 9 (E), and 12 M (F) groups showing progressive PNN loss. Scale 50 µm. (G–H) Bar graphs of CA2 PNN area coverage (WFA/PCP4 area) (G), and CA2 PNN (WFA) intensity (H) in 5XFAD and their age‐matched control mice at 3, 6, 9, and 12 M showing significant decreases at and after 6 M in 5XFAD; n = 7–8 slices from five mice per group in 3 M, n = 9–10 brain slices from five mice per group in 6 M, n = 9–10 brain slices from five mice per group in 9 M, n = 10–12 brain slices from five mice per group in 12 M. (I–J) PV neuron density (I), and NeuN neuron density (J) in CA2 area at 3 and 6 M 5XFAD mice compared to their age‐matched controls showing no difference between any groups; n = 8–9 brain slices from five mice per group in 3 M, n = 9–10 brain slices from five mice per group in 6 M. Bar data in G–H indicate mean ± SEM and dots represent data points. Two‐way ANOVA, Tukey's multiple comparisons test in G–H; unpaired two‐tailed t ‐test in I–J. * p < 0.05, ** p < 0.01, *** p < 0.001,**** p < 0.0001, and ns p > 0.05.

Article Snippet: We obtained 5XFAD mice (B6SJL‐Tg (APPSwFlLon, PSEN1*M146L*L286V) 6799Vas/ Mmjax, strain 034840‐JAX) from the Jackson Laboratory and backcrossed for five generations to C57Bl/6J (000664‐JAX) wild‐type (WT) mice to obtain an incipient congenic line on a C57Bl/6J genetic background for experiments.

Techniques: Disruption, Immunohistochemical staining, Expressing, Labeling, Control, Immunofluorescence, Two Tailed Test

Journal: Alzheimer's & Dementia

Article Title: Degradation of perineuronal nets in hippocampal CA2 explains the loss of social cognition memory in Alzheimer's disease

doi: 10.1002/alz.70813

Figure Lengend Snippet: Impairment of social memory in 5XFAD mice at 6 M age. (A) Schematic of the direct interaction social memory test. The test mouse (red dot) interacts with Novel 1 mouse (yellow dot) who becomes Familiar (yellow dot) in the second trial. In the following trial, the test mouse (red dot) interacts with another novel (Novel 2) mouse (green dot). (B) Graph showing time spent investigating by 3 M old 5XFAD and age‐matched control mice with Novel 1, Familiar, and Novel 2 mice suggesting normal social memory (two‐way repeated measures ANOVA, Tukey's multiple comparisons test ( n = 7) (control); 9 (5XFAD) mice; * p < 0.05, *** p < 0.001). (C–D) Bar graphs showing no significant change in difference scores, (C) Novel 1 – Familiar, and (D) Familiar − Novel 2, in 3 M 5XFAD mice compared to age‐matched control mice ( n = 7) (control); 9 (5XFAD) mice in (C–D), unpaired two‐tailed t ‐test. (E) Graph showing impaired social memory in 6 M 5XFAD mice compared to 6 M control mice ( n = 10) (control); 9 (5XFAD) mice, two‐way repeated measures ANOVA, Tukey's multiple comparisons test; * p < 0.05, **** p < 0.0001. (F) Bar graph showing significant change in difference scores (Novel 1 − Familiar) in 6 M 5XFAD mice compared to 6 M control mice ( n = 10) (control); 9 (5XFAD) mice; unpaired two‐tailed t ‐test; * p < 0.05. (G) Bar graph showing significant change in difference scores (Familiar − Novel 2) in 6 M 5XFAD mice compared to 6 M control mice ( n = 10) (control); 9 (5XFAD) mice; unpaired two‐tailed t ‐test, ** p < 0.01. (H) Schematic of novel object recognition task. The test mouse is familiarized with two objects (blue), followed by replacing one object with a novel object (green) in the next trial. (I) Bar graph showing significantly higher time spent with the novel object compared to familiar by both 6 M control and 5XFAD ( n = 11) (control); 17 (5XFAD) mice, two‐way ANOVA mixed‐effects Šídák's; ** p < 0.01. (J–K) Bar graph showing no significant difference in discrimination index (J), and distance moved (K) by both 6 M control and 5XFAD ( n = 11) (J), 17 (K) control; 17 (J), 22 (K) 5XFAD mice; unpaired two‐tailed t ‐test.

Article Snippet: We obtained 5XFAD mice (B6SJL‐Tg (APPSwFlLon, PSEN1*M146L*L286V) 6799Vas/ Mmjax, strain 034840‐JAX) from the Jackson Laboratory and backcrossed for five generations to C57Bl/6J (000664‐JAX) wild‐type (WT) mice to obtain an incipient congenic line on a C57Bl/6J genetic background for experiments.

Techniques: Control, Two Tailed Test

Journal: Alzheimer's & Dementia

Article Title: Degradation of perineuronal nets in hippocampal CA2 explains the loss of social cognition memory in Alzheimer's disease

doi: 10.1002/alz.70813

Figure Lengend Snippet: Transcriptomic analysis of age‐dependent CA2 area gene expression changes in 5XFAD mice. (A) Schematic of experimental design. We tested social memory in 3 and 7 M old control and 5XFAD mice followed by acute brain slicing and isolation of the CA2 area using a tissue puncher. CA2 area tissues were frozen fixed and processed for bulk RNA‐sequencing. (B) Volcano plot showing DEGs in the CA2 area in 3 M 5XFAD and age‐matched control mice. Genes with an adjusted p ‐value < 0.05 are shown in blue, genes with log 2 fold‐change > 1.00 in green, and genes meeting both criteria in red. Gray dots represent genes that do not meet either criterion. (C) Volcano plot showing DEGs in the CA2 area in 7 M 5XFAD and age‐matched control mice. Color coding is consistent with that in (B). (D–E) GO term circle plot highlighting the top 10 GO terms, sorted by adjusted p ‐value in (D) 3 M, and (E) 7 M, 5XFAD versus age‐matched control mice. Z‐scores indicate the directionality of regulation, with red reflecting upregulated genes and blue reflecting downregulated genes. (F, G) GO term circle plot of the top 10 GO terms related to PNNs and ECM, ranked by adjusted p ‐value. Z‐scores indicate the directionality of regulation, with red reflecting upregulated genes. (H–I) Heatmaps showing the expression levels of key genes involved in, (H) PNN synthesis, and (I) PNN degradation across all samples and experimental conditions. Expression values were normalized by z‐score transformation.

Article Snippet: We obtained 5XFAD mice (B6SJL‐Tg (APPSwFlLon, PSEN1*M146L*L286V) 6799Vas/ Mmjax, strain 034840‐JAX) from the Jackson Laboratory and backcrossed for five generations to C57Bl/6J (000664‐JAX) wild‐type (WT) mice to obtain an incipient congenic line on a C57Bl/6J genetic background for experiments.

Techniques: Gene Expression, Control, Isolation, RNA Sequencing, Expressing, Transformation Assay

Journal: Alzheimer's & Dementia

Article Title: Degradation of perineuronal nets in hippocampal CA2 explains the loss of social cognition memory in Alzheimer's disease

doi: 10.1002/alz.70813

Figure Lengend Snippet: The MMP blocker GM6001 delays social memory deficits and inhibits PNN degradation in 5XFAD mice. (A) Schematic of experimental design. We assessed social memory in 5 M 5XFAD and age‐matched control mice followed by i.p. GM6001 or vehicle injections for 1 month. Mice were again assessed for social memory after the injections and brains were collected for IHC. (B) Graph showing normal social memory in 5 M 5XFAD and age‐matched control mice ( n = 7) (control), 11 (5XFAD) mice; two‐way ANOVA, Tukey's multiple comparisons test; * p < 0.05, ** p < 0.01. (C–D) Bar graphs showing no significant change in difference scores, (C) Novel 1 – Familiar, and (D) Familiar − Novel 2, in 5 M 5XFAD mice compared to age‐matched control mice ( n = 7) (control); 11 (5XFAD) mice in (C–D), unpaired two‐tailed t ‐test. (E) Graph showing normal social memory in a month‐long GM6001‐injected 6 M 5XFAD mice whereas vehicle‐injected 6 M 5XFAD mice show disrupted social memory compared to the age‐matched control mice ( n = 14) (5XFAD + GM6001), 12 (5XFAD + vehicle) mice; two‐way ANOVA, Tukey's multiple comparisons test; * p < 0.05, **** p < 0.0001. (F–G) Bar graphs showing significant difference in difference scores (F) Novel 1 – Familiar, and (G) Familiar − Novel 2, in GM6001‐injected 6 M 5XFAD in 6 M compared to vehicle‐injected 6 M 5XFAD mice ( n = 14) (5XFAD + GM6001), 12 (5XFAD + vehicle) mice in (F–G); unpaired two‐tailed t ‐test; *** p < 0.001, **** p < 0.0001. (H) Representative confocal micrographs of PNN (WFA) immunofluorescence in CA2 area (PCP‐4) from 6 M 5XFAD mice injected with vehicle (top), or GM6001 (2nd from top), and 6 M Control (5XFAD‐) mice injected with vehicle (3rd from top) or GM6001 (bottom). Scale 50 µm. (I) Bar graphs showing significantly higher CA2 PNN expression in GM6001‐treated mice compared to vehicle‐treated 5XFAD mice whereas age‐matched GM6001‐treated or vehicle‐treated controls exhibited PNN expression comparable to 6 M GM6001‐treated 5XFAD mice (one‐way ANOVA, Tukey's multiple comparisons test; n = 4–5 mice per group; * p < 0.05, ** p < 0.01).

Article Snippet: We obtained 5XFAD mice (B6SJL‐Tg (APPSwFlLon, PSEN1*M146L*L286V) 6799Vas/ Mmjax, strain 034840‐JAX) from the Jackson Laboratory and backcrossed for five generations to C57Bl/6J (000664‐JAX) wild‐type (WT) mice to obtain an incipient congenic line on a C57Bl/6J genetic background for experiments.

Techniques: Control, Two Tailed Test, Injection, Immunofluorescence, Expressing

Journal: iScience

Article Title: Cross-species comparative hippocampal transcriptomics in Alzheimer’s disease

doi: 10.1016/j.isci.2023.108671

Figure Lengend Snippet: Shared DEGs among EOAD, LOAD, and mouse models of AD Schematic summary of samples used and analysis workflow to obtain DEGs. (A) Venn diagram showing DEGs overlap between EOAD (B) and LOAD (C) with 5xFAD, APP/PS1, and hAβ-KI mice. Mosaic plot of hAβ-KI, 5xFAD, and APP/PS1 overlap (D) Mosaic plot of EOAD-model (left) and model-EOAD (right) DEGs overlap with 5xFAD, APP/PS1, and hAβ-KI mice (E) Mosaic plot of LOAD-model (left) and model-LOAD (right) DEGs overlap with 5xFAD, APP/PS1, and hAβ-KI mice (F) PPI network of DEGs exclusively shared between hAβ-KI mice and LOAD (G) or EOAD (H) patients. Mosaic plot of EOAD, LOAD, or MS DEGs overlaps with 5xFAD mice (I) Mosaic plot of EOAD, LOAD, or MS DEGs overlaps with APP/PS1 mice (J) Mosaic plot of EOAD, LOAD, or MS DEGs overlaps with hAβ-KI mice (K) The size of red and yellow boxes reflects the proportion of overlapping and non-overlapping DEGs, respectively. Pearson’s Chi−squared test with Yates’ continuity correction was applied for the mosaic plot analysis. EOAD, early-onset Alzheimer’s disease; LOAD, late-onset Alzheimer’s disease; MS, multiple sclerosis; hAβ-KI, humanized amyloid-β knock-in. Genes with unadjusted p value <0.05 were considered as DEGs.

Article Snippet: Moreover, considering the total of DEGs identified in the mouse models as reference (model-disease overlap), the intersection of DEGs between 5xFAD and EOAD was significantly higher (14%) than the overlap between both APP/PS1 (10.9%; Chi-square adjusted p value = 0.0061) or hAβ-KI (10.7%; Chi-square adjusted p value = 0.0054) with EOAD ( E - right).

Techniques: Knock-In

Journal: iScience

Article Title: Cross-species comparative hippocampal transcriptomics in Alzheimer’s disease

doi: 10.1016/j.isci.2023.108671

Figure Lengend Snippet: Number of differentially expressed genes by each brain cell type

Article Snippet: Moreover, considering the total of DEGs identified in the mouse models as reference (model-disease overlap), the intersection of DEGs between 5xFAD and EOAD was significantly higher (14%) than the overlap between both APP/PS1 (10.9%; Chi-square adjusted p value = 0.0061) or hAβ-KI (10.7%; Chi-square adjusted p value = 0.0054) with EOAD ( E - right).

Techniques:

Journal: iScience

Article Title: Cross-species comparative hippocampal transcriptomics in Alzheimer’s disease

doi: 10.1016/j.isci.2023.108671

Figure Lengend Snippet: GOBP overlap among EOAD, LOAD, and mouse models of AD Schematic summary of samples used and analysis workflow to obtain GOBPs. (A) Venn diagram of GOBPs intersections in EOAD (B) and LOAD (C) with 5xFAD, APP/PS1, and hAβ-KI mice. Mosaic plot of hAβ-KI-5xFAD and -APP/PS1 GOBP overlap (D) Mosaic plot of EOAD-model (left) and model-EOAD (right) GOBP overlap with 5xFAD, APP/PS1, and hAβ-KI mice (E) Mosaic plot of LOAD-model (left) and model-LOAD (right) GOBP overlap with 5xFAD, APP/PS1, and hAβ-KI mice (F) Mosaic plot of EOAD, LOAD, or MS GOBP overlap with 5xFAD mice (G) Mosaic plot of EOAD, LOAD, or MS GOBP overlap with APP/PS1 mice. (H) Mosaic plot of EOAD, LOAD, or MS GOBP overlap with hAβ-KI mice. (I) The size of the red and yellow boxes reflects the proportion of overlapping and non-overlapping GOBPs, respectively. Pearson’s Chi−squared test with Yates' continuity correction was applied for the mosaic plot analysis. EOAD, early-onset Alzheimer’s disease; LOAD, late-onset Alzheimer’s disease; MS, multiple sclerosis; hAβ-KI, humanized amyloid-β knock-in.

Article Snippet: Moreover, considering the total of DEGs identified in the mouse models as reference (model-disease overlap), the intersection of DEGs between 5xFAD and EOAD was significantly higher (14%) than the overlap between both APP/PS1 (10.9%; Chi-square adjusted p value = 0.0061) or hAβ-KI (10.7%; Chi-square adjusted p value = 0.0054) with EOAD ( E - right).

Techniques: Knock-In

Journal: iScience

Article Title: Cross-species comparative hippocampal transcriptomics in Alzheimer’s disease

doi: 10.1016/j.isci.2023.108671

Figure Lengend Snippet: Functional enrichment analysis of KEGG terms in mouse models of AD and human disease Venn diagrams showing the KEGGs overlap among EOAD (A) and LOAD (B) patients with 5xFAD, APP/PS1, and hAβ-KI mice. Mosaic plot of 5xFAD, APP/PS1, and hAβ-KI KEGGs overlap with EOAD (C) or LOAD (D). Dual-plots showing the up- (red) and down- (blue) regulated KEGGs for EOAD (E), LOAD (F), 5xFAD (G), APP/PS1 (H), and hAβ-KI (I). The size of the red and yellow boxes reflects the proportion of overlapping and non-overlapping KEGGs, respectively. Pearson’s Chi−squared test with Yates' continuity correction was applied for the mosaic plot analysis. The pathways in the dual-plots are represented in the Y axis; the left and the right part of the X axis corresponds to the number of genes enriched in the KEGG terms and their q-value (gray bars), respectively. EOAD, early-onset Alzheimer’s disease; LOAD, late-onset Alzheimer’s disease; MS, multiple sclerosis; hAβ-KI, humanized amyloid-β knock-in.

Article Snippet: Moreover, considering the total of DEGs identified in the mouse models as reference (model-disease overlap), the intersection of DEGs between 5xFAD and EOAD was significantly higher (14%) than the overlap between both APP/PS1 (10.9%; Chi-square adjusted p value = 0.0061) or hAβ-KI (10.7%; Chi-square adjusted p value = 0.0054) with EOAD ( E - right).

Techniques: Functional Assay, Knock-In

Journal: iScience

Article Title: Cross-species comparative hippocampal transcriptomics in Alzheimer’s disease

doi: 10.1016/j.isci.2023.108671

Figure Lengend Snippet: Master regulator analysis of mouse models of AD and human disease Venn diagrams of EOAD (A) or LOAD (B) with 5xFAD, APP/PS1, and hAβ-KI MRs overlap. Tile-plot of regulons significantly enriched with DEGs in EOAD, LOAD, 5xFAD, APP/PS1, and hAβ-KI (only regulatory units significant in ≥4 contexts are represented). (C) Tile-plot showing the activation state of the transcription factors acting as MR in EOAD, LOAD, 5xFAD, APP/PS1, and hAβ-KI. (D) Venn diagram showing the overlap between previously MR candidates found by Vargas et al. and the current study. (E) EOAD, early-onset Alzheimer’s disease; LOAD, late-onset Alzheimer’s disease; hAβ-KI, humanized amyloid-β knock-in; ns, non-significant.

Article Snippet: Moreover, considering the total of DEGs identified in the mouse models as reference (model-disease overlap), the intersection of DEGs between 5xFAD and EOAD was significantly higher (14%) than the overlap between both APP/PS1 (10.9%; Chi-square adjusted p value = 0.0061) or hAβ-KI (10.7%; Chi-square adjusted p value = 0.0054) with EOAD ( E - right).

Techniques: Activation Assay, Knock-In

Journal: iScience

Article Title: Cross-species comparative hippocampal transcriptomics in Alzheimer’s disease

doi: 10.1016/j.isci.2023.108671

Figure Lengend Snippet:

Article Snippet: Moreover, considering the total of DEGs identified in the mouse models as reference (model-disease overlap), the intersection of DEGs between 5xFAD and EOAD was significantly higher (14%) than the overlap between both APP/PS1 (10.9%; Chi-square adjusted p value = 0.0061) or hAβ-KI (10.7%; Chi-square adjusted p value = 0.0054) with EOAD ( E - right).

Techniques: Reverse Transcription, Gene Expression, Microarray, Software

Journal: bioRxiv

Article Title: Vascular smooth muscle cell loss, but not neuroinflammation, drives cerebrovascular reactivity impairment in Alzheimer’s disease

doi: 10.64898/2025.12.01.691642

Figure Lengend Snippet: Comparisons of MRI measurements across young wild-type (WT; n = 8, 5F/3M), young 5xFAD ( n = 10, 5F/5M), older WT ( n = 12, 9F/3M), and older 5xFAD ( n = 16, 7F/9M) mice, including brain volume (a), cerebral blood flow (CBF; b), global cerebrovascular reactivity (CVR; c), regional mean CBF maps (d), and isocortical and hippocampal CVR (e). Red and blue dots denote female and male mice, respectively. (f) Representative immunofluorescent images of α-SMA and Collagen IV staining. Scale bars: 1 mm (whole-slice views) and 50 µm (regional views). (g) Comparisons of α-smooth muscle actin (α-SMA) coverage index across young WT ( n = 5, 3F/2M), young 5xFAD ( n = 5, 3F/2M), older WT ( n = 5, 3F/2M), and older 5xFAD ( n = 5, 1F/4M) mice, which is defined as the ratio of the co-localized α-SMA and Collagen IV-positive area (α-SMA + Col-IV + ) to the total Collagen IV-positive area (Col-IV + ). For group-wise comparisons: * p < 0.05; ** p < 0.01; n.s., not significant.

Article Snippet: The 5xFAD line (Jackson Laboratory, stock #034848) is an APP/PS1 double-transgenic model carrying five familial AD mutations: the Swedish (K670N/M671L), Florida (I716V), and London (V717I) mutations in APP, and the M146L and L286V mutations in PSEN1.

Techniques: Staining

Journal: bioRxiv

Article Title: Vascular smooth muscle cell loss, but not neuroinflammation, drives cerebrovascular reactivity impairment in Alzheimer’s disease

doi: 10.64898/2025.12.01.691642

Figure Lengend Snippet: (a, b) Slice-by-slice and regional comparisons of immunofluorescent staining for amyloid-β (Aβ) in representative young wild-type (WT), young 5xFAD, older WT, and older 5xFAD mice. Scale bars: 1 mm (whole-slice views) and 50 µm (regional views). (c) Amyloid-β coverage index, defined as the ratio of 6E10-positive area (6E10 + ) to the total regional area, in the isocortex and hippocampus across the four groups. Red and blue dots represent female and male mice, respectively. Error bars indicate standard deviation. (d) Vascular amyloid coverage index, defined as the ratio of co-localized 6E10- and Collagen IV-positive area (6E10 + Col-IV + ) to the total 6E10-positive area (6E10 + ), in the isocortex and hippocampus. For group-wise comparisons: * p < 0.05; *** p < 0.001.

Article Snippet: The 5xFAD line (Jackson Laboratory, stock #034848) is an APP/PS1 double-transgenic model carrying five familial AD mutations: the Swedish (K670N/M671L), Florida (I716V), and London (V717I) mutations in APP, and the M146L and L286V mutations in PSEN1.

Techniques: Staining, Standard Deviation

Journal: bioRxiv

Article Title: Vascular smooth muscle cell loss, but not neuroinflammation, drives cerebrovascular reactivity impairment in Alzheimer’s disease

doi: 10.64898/2025.12.01.691642

Figure Lengend Snippet: Comparisons of MRI measurements across pre-saline (Pre-Sal), post-saline (Post-Sal), pre-LPS, and post-LPS groups, including brain volume (a), cerebral blood flow (CBF; b), global cerebrovascular reactivity (CVR; c), regional mean CBF maps (d), and hippocampal CBF and CVR (e). Red and blue dots denote female and male mice, respectively. Dots connected by a line correspond to the same mouse. (f) Representative immunofluorescent images showing ionized calcium-binding adaptor molecule 1 (Iba1, red) and CD68 (green) staining in the hippocampus. Scale bars: 50 µm. Both the LPS-induced neuroinflammation and 5xFAD models are shown for comparison. (g) Comparisons of the CD68 coverage index across pre- (Pre-Sal and Pre-LPS; n = 5, 2F/3M), post-Sal ( n = 5, 3F/2M), and post-LPS ( n = 5, 3F/2M) groups in the hippocampus and isocortex. Error bars indicate standard deviation. The CD68 coverage index is defined as the ratio of the co-localized Iba1- and CD68-positive area (Iba1 + CD68 + ) to the total Iba1-positive area (Iba1 + ). (h) Comparisons of the CD68 coverage index across young WT ( n = 5, 3F/2M), young 5xFAD ( n = 5, 3F/2M), older WT ( n = 5, 3F/2M), and older 5xFAD ( n = 5, 1F/4M) mice.

Article Snippet: The 5xFAD line (Jackson Laboratory, stock #034848) is an APP/PS1 double-transgenic model carrying five familial AD mutations: the Swedish (K670N/M671L), Florida (I716V), and London (V717I) mutations in APP, and the M146L and L286V mutations in PSEN1.

Techniques: Saline, Binding Assay, Staining, Comparison, Standard Deviation

Journal: bioRxiv

Article Title: Functional border-associated macrophages limit Alzheimer’s Disease progression

doi: 10.64898/2026.01.31.703045

Figure Lengend Snippet: A, Volcano plot with all significant differentially expressed genes between BAM2 (Cluster 4 in ) and microglia (Clusters 0,1,2,5,6) (n=4 WT mice 5-6 Mo, 2M, 2F). B , All KEGG pathways enriched in BAM2 > microglia. C , Similar to ( A ) but only for all mapped genes associated with an AD risk allele (MONDO_0004975). D , Whisker plots surrounding whisker plots with median, 25% and 75% depicting the distribution of AD enrichment score per cell within total clusters, microglia, or BAM. E , Experimental design for rhodamine dextran i.v. administration; F , representative histograms for each brain macrophage subtype; G , quantification of % of rhodamine positive cells; H , quantification of median fluorescence intensity (MFI) for intravenous rhodamine endocytosis assay (n=4 WT mice). I-L As with ( E-H ) but for an experiment in which rhodamine dextran was injected into CSF via cisterna magna (n=6 WT mice). M-P, As with ( E-H ) but for an experiment in which single cell suspensions were incubated with fluorescein amidite (FAM) tagged Aβ 1-42 oligomers for 15 minutes (n=5 WT mice). Q-S, As with ( E-G) but for an experiment in which Methoxy-X04 was injected intraperitoneally (IP) 3 hours prior to the generation of single cell suspensions (n=5 5xFAD mice + WT mouse for control comparison). T, Representative image with white arrows depicting LYVE1 + CD206 + BAM2 in human post-mortem superior frontal gyrus colocalized with Aβ deposited around CD31 + vasculature. Scale bar=50 µm. U , Analysis of the change in expression of human AD GWAS genes comparing AD cells to healthy cells within the same cluster (genes from MONDO_0004975, see Materials and methods section for datasets used). A Hampel filter was used to calculate threshold values and colored dots represent clusters enriched or depleted in change from healthy to AD. ( D ) Wilcoxon rank sum test with continuity correction. ( G-H, K-L, O-P, S ) One-way matched ANOVA with Sidak’s multiple comparisons test. *P<.05, **P<.01, ***P<.001, ****P<.0001. Bar plots with connecting lines represent populations from the same animal. AD= Alzheimer’s Disease.

Article Snippet: C57BL/6J (000664), LySM Cre (04781), and Csf1r Cre (021024), 5xFAD (034840) strains were purchased from the Jackson Laboratories.

Techniques: Whisker Assay, Fluorescence, Endocytosis Assay, Injection, Single Cell, Incubation, Control, Comparison, Expressing

Journal: bioRxiv

Article Title: Functional border-associated macrophages limit Alzheimer’s Disease progression

doi: 10.64898/2026.01.31.703045

Figure Lengend Snippet: A, Schematic of oligomeric soluble Aβ 1-42 endocytosis assay (as in ); B , representative histograms of Aβ 1-42 positivity (right of dotted line) for BAM2 (top) and microglia (bottom) with respective quantification of fold change in 3-4 mo mice (n= 5 WT, 5 5xFAD mice); c , as with ( B ) but for 7-9 mo mice. WT avg (n=9 WT, 14 5xFAD mice). D-E , Quantification of ( D) BAM and ( E ) microglia as percentage of CD64 + (total) macrophages (n=6 WT, 7 5xFAD mice); ( F) representative zebra plots from 7-9 mo mice from WT (top, black) and 5xFAD (bottom, red) mice with percentage of each brain macrophage subtype; G-H, as with ( D-E ) but for 7-9 mo mice (n=8 WT, 6 5xFAD mice); I , UMAP of aggregated human brain macrophages from healthy control and AD patients highlighting microglia clusters (light blue box) and BAM clusters (dark blue box). J , UMAP of BAM subclusters. K , BAM1/BAM2 fraction per patient from the ROSMAP dataset (n=151 healthy, 81 AD patients). L , Representative micrographs of CODEX-stained AD and age-matched control superior frontal gyrus for BAM2 maker LYVE1, CD206, vasculature maker CD31 and CD34, and Aβ along with large vessel ROIs. M-O , quantification of the ( M ) % of vessel associated cells that are BAM2 (LYVE1 + ), (n=6 healthy, 14 AD), ( N ) BAM1 (CD206 + LYVE1 - ), (n=6 healthy, 14 AD) and ( O ) the ratio of BAM2/BAM1 (LYVE1 + /CD206 + LYVE1 - ) (n=6 healthy, 11 AD). P,Q , Forrest plots demonstrating correlation between ( P ) BAM2/BAM1 enrichment score and different AD-clinical measures. ( Q ) % of BAM2s of total macrophages and different AD-clinical measures. ( B,C,D,G ) Two-way matched ANOVA with Sidak’s multiple comparisons test. ( E,H,K,M,N,O ) Two-tailed unpaired t-test. *P<.05, **P<01, ****P<.0001. ( i ) Scale bars=500µm (left), 100 µm (middle/right). Fold change normalized to 1 (as average) includes individual values that are not 1; these derive from littermates with the same genotype and slightly different individual values. UMAP= uniform manifold approximation and projection.

Article Snippet: C57BL/6J (000664), LySM Cre (04781), and Csf1r Cre (021024), 5xFAD (034840) strains were purchased from the Jackson Laboratories.

Techniques: Endocytosis Assay, Control, Staining, Two Tailed Test

Journal: bioRxiv

Article Title: Functional border-associated macrophages limit Alzheimer’s Disease progression

doi: 10.64898/2026.01.31.703045

Figure Lengend Snippet: A, IBA1 + (red) microglia staining and association with X-34 staining (cyan) of compacted amyloid in sections of BAM2-depleted Csf1R Cre/+ Maf F/F 5xFAD mice and Maf F/F 5xFAD controls from similar regions of cortex and hippocampus. B-C , quantification IBA1 area normalized to amyloid area in ( B ) cortex and ( C ) hippocampus (n=4 Maf F/F 5xFAD, 5 Csf1R Cre/+ Maf F/F 5xFAD mice). Two-tailed unpaired student’s t-test. Scale bar=100µm low magnification images, 50µm for high magnification insets.

Article Snippet: C57BL/6J (000664), LySM Cre (04781), and Csf1r Cre (021024), 5xFAD (034840) strains were purchased from the Jackson Laboratories.

Techniques: Staining, Two Tailed Test

Journal: bioRxiv

Article Title: Functional border-associated macrophages limit Alzheimer’s Disease progression

doi: 10.64898/2026.01.31.703045

Figure Lengend Snippet: A , Representative micrographs of X-34 amyloid staining in cortex and hippocampus in 3-5 month-old Cre + 5xFAD (BAM2 depleted) and Cre - (BAM2 sufficient) 5xFAD littermates (top) with regions of interest (ROIs, numbered boxes below) depicting pial and penetrating CD31 + blood vessels in cortex and the hippocampal artery in hippocampus. B , Quantification of X-34 cortical area fraction normalized to littermates at 3-5 mo (n= 11 Cre - , 12 Cre + mice) and 7-9 mo (n=7 Cre - , 6 Cre + mice). C , as with ( B ) but for hippocampus. D , Quantification of the percentage of pial and penetrating blood vessel area covered with CAA, presented as fold change normalized to littermates at 3-5 mo (n= 6 Cre - , 6 Cre + mice) and 7-9mo (n=7 Cre - , 6 Cre + mice) mice. E-H , ELISA based quantification of 3 mo whole brain ( E) soluble Aβ 1-40 (n= 6 Cre - , 7 Cre + mice), ( F) soluble Aβ 1-42 (n= 6 Cre - , 7 Cre + mice), ( G ) insoluble Aβ 1-40 (n= 5 Cre - , 4 Cre + mice), and ( H ) insoluble Aβ 1-42 (n= 6 Cre - , 6 Cre + mice). I , representative micrographs of LAMP1 staining in cortex and hippocampus in 3-5 mo Cre + and Cre - littermates, along with high magnification ROIs (numbered boxes below) in sections. Insets 1 and 2 show sections from Cre + mice depicting LAMP1 localization around X-34 + plaque (Inset 1) and vasculature (Inset 2, showing salient CAA). Inset 3 depicts CA3 granular cell layer in hippocampus from Cre + mice in which LAMP1 staining appears in gaps in NeuN+ neuron staining (red arrows). J , Quantification of cortical LAMP1 area fraction at 3-5mo (n=10 Cre - , 9 Cre + mice) and 7-9mo (n=7 Cre - , 7 Cre + mice); K , as with ( J ) but for hippocampal LAMP1 at 3-5mo (n=9 Cre - , n=12 Cre + mice), and 7-9mo (n= 6 Cre - , 5 Cre + mice). L-M , neuronal loss in Cre + mice. ( L ) representative micrograph depicting NeuN+ neuron staining in cortical layers from 9 mo Cre + and Cre - littermates; M , quantification of L5 neuron number/area normalized to littermates at 3-5 mo (n=6 Cre - , 6 Cre + mice) and 7-9 mo (n=8 Cre - , 6 Cre + ). N-O, behavioral tests. (N), schematic of contextual fear conditioning paradigm; ( O) , quantification of freezing proportion in first 5 minutes of memory retrieval (blue=male (M), red=female (F), n=8 WT M , 7WT F , 11 Cre + M , 10 Cre + F , 11 5xFAD M , 9 5xFAD F , 9 Cre + 5xFAD M , 10 Cre + 5xFAD F ). Main effect of sex (F(1,67)= 10.133, p = 0.002) and Cre status (F(1,67)= 10.012, p = 0.002). P , Schematic of Barnes maze test along with longitudinal design including trials, days, and age (months). Q-S Quantification, Cre x 5xFAD x Age interaction, and 5xFAD main effect respectively of ( Q ) average distance traveled (F(6,1855) = 2.978, p = 0.007) and (F(1,1855) = 6.200, p = 0.013); ( R ) committed (F(6,1856) = 3.267, p = 0.003) and (F(1,1856) = 12.550, p < .001); and ( S ) latency (s) to locate the escape hole for each day and age (F(6,1856) = 2.098, p = 0.051) and (F(1,1856) = 8.447, p = 0.004)). See Materials and Methods for n values). ( A, I ) White dotted line represents hippocampal area. ( B-D, J-K, M, O ) Two-way ANOVA with Sidak’s multiple comparisons test, ( E-H ) two-tailed unpaired t-test, ( Q-S ) Linear Mixed Effects Model with Sidak’s multiple comparisons test. Significant pairwise comparisons within time points indicated by shapes associated with respective genotypes in the panel legend. *P<.05,**P<.01, ***P<.001, ****p<.0001. ( A,I,L ) Scale bars=200µm large images, 50µm high mag ROIs. Fold change normalized to 1 (as average) includes individual values that are not 1; these derive from littermates with the same genotype and slightly different individual values.

Article Snippet: C57BL/6J (000664), LySM Cre (04781), and Csf1r Cre (021024), 5xFAD (034840) strains were purchased from the Jackson Laboratories.

Techniques: Staining, Enzyme-linked Immunosorbent Assay, Two Tailed Test

Journal: bioRxiv

Article Title: Functional border-associated macrophages limit Alzheimer’s Disease progression

doi: 10.64898/2026.01.31.703045

Figure Lengend Snippet: A . Representative micrographs of X-34 amyloid staining in cortex and hippocampus in 7-9 month old Cre + (BAM2 depleted) and Cre - 5xFAD littermates with regions of interest (ROIs, green boxes) depicting CAA in pial and penetrating CD31 + blood vessels in cortex. B. As with A but for LAMP1 neurodystrophy marker. Scale bars=200µm large images, 50µm high mag ROIs.

Article Snippet: C57BL/6J (000664), LySM Cre (04781), and Csf1r Cre (021024), 5xFAD (034840) strains were purchased from the Jackson Laboratories.

Techniques: Staining, Marker

Journal: bioRxiv

Article Title: Functional border-associated macrophages limit Alzheimer’s Disease progression

doi: 10.64898/2026.01.31.703045

Figure Lengend Snippet: A, Heatmap of relative Lys2 ( LysM ) expression per cell in clusters detailed in Fig. S1b . B , Representative contour plots of brain macrophage populations (BAM2, top box; BAM1, right box; MG, bottom left box) with quantification of percentage of CD64 + gated cells. Note an increase in percentage of BAM1, which does not reflect a conversion of BAM2 into BAM1, but a rebound of BAM2, which are repopulated from monocytes . C , Representative micrographs depicting X-34 amyloid staining and CD31 + endothelial cells in LysM Cre/+ Maf F/F 5xFAD and Maf F/F 5xFAD mice with high magnification insets depicting prominent CAA in LysM Cre/+ Maf F/F 5xFAD mice. D-F , Quantification of ( D ) CAA % blood vessel area, ( E ) cortical, and ( F ) hippocampal X-34 % area normalized to same sex littermate controls. Mean+/- S.E.M. Scale bars=200µm large images, 50µm insets.

Article Snippet: C57BL/6J (000664), LySM Cre (04781), and Csf1r Cre (021024), 5xFAD (034840) strains were purchased from the Jackson Laboratories.

Techniques: Expressing, Staining

Journal: bioRxiv

Article Title: Functional border-associated macrophages limit Alzheimer’s Disease progression

doi: 10.64898/2026.01.31.703045

Figure Lengend Snippet: A , Contextual Fear Conditioning experiment recall ( from ) binned by 1-minute intervals. Significant 5xFAD x Minute interaction (Huynh-Feldt: F(6.982,432.880) = 2.397, p=0.021) and main effect of Cre (F(1,62) = 7.101, p=0.01). See main figure legend for n values. B-G , ( B ) Barnes Maze experiment ( from ) with ( C ) average number of trials frozen per animal by day, ( D ) avg trials with hole found by day (Cre x 5xFAD x Day interaction (F(6,1858) = 3.766, p < 0.001) ( E ) avg Distance by trail, ( F ) avg Errors by trial, and ( G ) avg Latency per animal by trial. Two-way repeated measures ANOVA with Sidak’s multiple comparisons test. See Materials and Methods for n values. Significant pairwise comparisons and select p values within time points indicated by shapes associated with respective genotypes in the panel legend. Linear Mixed Effects Model revealed a main effect of Trial across Barnes Maze performance measures: ( D ) F(3,1856) = 17.854, p<0.001), ( E ) F(3,1856) = 13.585, p<0.001), ( F ) F(3,1856) = 9.188, p<0.001). ( C ) Sidak comparisons also revealed that Cre + 5xFAD mice had a reduced ability to find the escape hole across days (all p<0.05), unlike all other groups. *P<.05,**P<.01, ***P<.001. TP=Time Point (months), D=days.

Article Snippet: C57BL/6J (000664), LySM Cre (04781), and Csf1r Cre (021024), 5xFAD (034840) strains were purchased from the Jackson Laboratories.

Techniques:

Journal: bioRxiv

Article Title: Functional border-associated macrophages limit Alzheimer’s Disease progression

doi: 10.64898/2026.01.31.703045

Figure Lengend Snippet: A , Schematic of TMRE assay. B , Representative histogram of TMRE staining in macrophage subtypes along with TMRE level delineation (dotted lines). C-D , quantification of ( c ) TMRE MFI per cell type and ( D ) percentage of TMRE hi . E , representative histograms of TMRE populations within BAM2 (top) and microglia (bottom) from 3-4 mo mice, along with respective quantification of percentage of TMRE hi population normalized to avg WT levels for each cell type (n=5 WT, 5 5xFAD mice). F, as with ( E ) but for 7-9 mo mice (n=7 WT, 7 5xFAD mice). G , Schematic of experiment in which TMRE and Aβ 1-42 were co-incubated prior to flow cytometry. H , representative heatmap comparing TMRE (x axis, high vs low/negative) to Aβ positivity (y axis, positive vs negative) in BAM2 cells, and respective quantification of Aβ positivity within each TMRE population (n=4 WT, 5 5xFAD mice). I , Schematic of experiment in which single cell suspensions were incubated with FCCP or DMSO (control) prior to Aβ 1-42 and TMRE. J-K , Quantification of ( J ) mean fluorescence intensity (MFI) of TMRE and ( K ) intracellular FAM-Aβ 1-42 for each cell type in experiment I, with microglia MFI magnified in inset. L , Quantification of percentage of BAM2 and microglia positive for mitochondrial superoxide (MitoSOX Red) from 7-9 mo mice (n=4 WT, 4 5xFAD mice). M , Quantification of percentage of BAM2 and microglia positive for cytoplasmic ROS using CellROX Deep Red probe in 7-9 mo mice (n=5 WT, 5 5xFAD mice). N , Quantification of percentage of senescent BAM2 and microglia using CellEvent TM Senescence Green probe in 7-9 mo mice (n=5 WT, 5 5xFAD mice) with microglia percentage magnified in inset. O-Q , ( O ) Schematic of experiment multiplexing Methoxy-X04, CellROX, and CellEvent TM with respective quantification ( P ) Percentage of Methoxy-X04 positive BAM2 cells correlative to CellROX positive or negative BAM2, and ( Q ) Percentage of Senescent (CellEvent TM positive) BAM2 cells correlative with CellROX positive or negative BAM2 from 7-9 mo mice (n= 5 5xFAD mice). ( C,D ) One-way matched ANOVA with Sidak’s post hoc test. ( E-F, J-N ) Two-way matched ANOVA with Sidak’s multiple comparisons test. ( P-Q ) Two-tailed paired t-test. ( K,N insets) Two-tailed unpaired t-test *P<.05, **P<.01, ***P<.001, ****P<.0001. ( C-D, H, P-Q ) connecting lines=same animal. βgal= β-galactosidase, ns= not significant, ROS/ROX= reactive oxygen species, SOX=superoxide, FCCP= carbonyl cyanide-p-trifluoromethoxyphenylhydrazone, TMRE= tetramethylrhodamine, ethyl ester. Fold change normalized to 1 (as average) includes individual values that are not 1; these derive from littermates with the same genotype and slightly different individual values.

Article Snippet: C57BL/6J (000664), LySM Cre (04781), and Csf1r Cre (021024), 5xFAD (034840) strains were purchased from the Jackson Laboratories.

Techniques: Staining, Incubation, Flow Cytometry, Single Cell, Control, Fluorescence, Multiplexing, Two Tailed Test

Journal: Neurotherapeutics

Article Title: Microglial GM3 accumulation impairs Aβ phagocytic activity and promotes neuroinflammation in Alzheimer's disease

doi: 10.1016/j.neurot.2026.e00851

Figure Lengend Snippet: Elevated expression of Hexa and Hexb in the brains of 5xFAD mice and human patients with AD. (A) Hexa and Hexb , but not St3gal5 , mRNA levels are increased in the cerebral cortex of 5xFAD animals compared to their wild-type littermates (n = 3–5 per group, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001, one-way ANOVA and Tukey's post hoc analysis). (B) Hexa and Hexb , but not St3gal5 , mRNA levels are increased in the hippocampus of 5xFAD animals compared to their wild-type littermates (n = 3–4 per group, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001, one-way ANOVA and Tukey's post hoc analysis). (C) Normalized expression levels of ST3GAL5 , HEXA , and HEXB in the human entorhinal cortex ( GSE48350 ). HEXA expression is significantly increased in female patients with AD compared to healthy female controls (∗∗p < 0.01, Student's t-test). (D) Normalized expression levels of ST3GAL5 , HEXA , and HEXB in the human hippocampus ( GSE48350 ). HEXA expression is significantly increased in female patients with AD compared to healthy female controls (∗∗∗p < 0.001, Student's t-test).

Article Snippet: 5xFAD animals (C57BL/6J background, stock no: 034848) were purchased from the Jackson Laboratory and maintained in temperature-, light-, and humidity-controlled conditions with ad libitum access to food and water.

Techniques: Expressing

Journal: Neurotherapeutics

Article Title: Microglial GM3 accumulation impairs Aβ phagocytic activity and promotes neuroinflammation in Alzheimer's disease

doi: 10.1016/j.neurot.2026.e00851

Figure Lengend Snippet: Hexa and Hexb protein expression is selectively elevated in 5xFAD and APP NLGF mice but not in JNPL3 tauopathy mice. (A) Hexa and Hexb protein expression, but not St3gal5, was significantly increased in the cerebral cortex of 5xFAD mice. Notably, female 5xFAD mice exhibited higher Hexa and Hexb expression levels than their male counterparts (∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, one-way ANOVA and Tukey's post hoc analysis). (B) Hexa and Hexb protein levels, but not St3gal5, progressively increased in the hippocampus of female 5xFAD mice as the disease advanced (∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001, one-way ANOVA and Tukey's post hoc analysis). (C) Hexa and Hexb protein levels, but not St3gal5, progressively increased in the cortex of APP NLGF mice with disease progression (∗∗∗p < 0.001, ∗∗∗∗p < 0.0001, one-way ANOVA and Tukey's post hoc analysis, circles represent males; triangles represent females). (D) The expression levels of St3gal5, Hexa, and Hexb remained consistent across all groups during disease progression in the JNPL3 tauopathy mouse model.

Article Snippet: 5xFAD animals (C57BL/6J background, stock no: 034848) were purchased from the Jackson Laboratory and maintained in temperature-, light-, and humidity-controlled conditions with ad libitum access to food and water.

Techniques: Expressing, Biomarker Discovery

Journal: Neurotherapeutics

Article Title: Microglial GM3 accumulation impairs Aβ phagocytic activity and promotes neuroinflammation in Alzheimer's disease

doi: 10.1016/j.neurot.2026.e00851

Figure Lengend Snippet: Hexa and Hexb expression is predominantly upregulated in microglial cells of the cerebral cortex and hippocampus in 5xFAD mice. (A) The bar graph shows the average expression levels of Hexa and Hexb across different cell types in the cerebral cortex. The data were obtained from the Gene Expression Omnibus (GEO) public repository ( GSE140399 ). (B) The bar graph shows Hexa expression across different cell types in the cerebral cortex under both wild-type and 5xFAD conditions (∗p < 0.05, ∗∗p < 0.01, Mann-Whitney U test). (C) The bar graph shows Hexb expression across different cell types in the cerebral cortex under both wild-type and 5xFAD conditions (∗∗p < 0.01, ∗∗∗∗p < 0.0001, Mann-Whitney U test). (D) The bar graph displays the average expression levels of Hexa and Hexb across different cell types in the hippocampus. The data were obtained from the GEO public repository ( GSE140399 ). (E) The bar graph displays Hexa expression across different cell types in the hippocampus under both wild-type and 5xFAD conditions (∗p < 0.05, ∗∗p < 0.01, ∗∗∗∗p < 0.0001, Mann-Whitney U test). (F) The bar graph displays Hexb expression across different cell types in the hippocampus under both wild-type and 5xFAD conditions (∗∗p < 0.01, ∗∗∗∗p < 0.0001, Mann-Whitney U test). (G) Representative images of Iba1 and GM3 staining in the cerebral cortex from male and female wild-type and 5xFAD mice. Quantification of GM3 + /Iba1 + signal reveals that 5xFAD animals exhibit higher GM3 levels in microglia compared to their wild-type littermates. Additionally, female 5xFAD mice show increased GM3 + /Iba1 + signal relative to male 5xFAD mice (∗∗∗∗p < 0.0001, one-way ANOVA and Tukey's post hoc analysis, scale bar = 50 μm, inset scale bar = 20 μm). (H) Representative images of Iba1 and GM3 staining in the hippocampus from male and female wild-type and 5xFAD mice. GM3 + /Iba1 + signal quantification indicates that GM3 levels in microglia are elevated in 5xFAD mice relative to wild-type controls. Moreover, female 5xFAD mice display a greater GM3 + /Iba1 + signal than their male counterparts (∗p < 0.05, ∗∗p < 0.01, ∗∗∗∗p < 0.0001, one-way ANOVA and Tukey's post hoc analysis, scale bar = 50 μm, inset scale bar = 20 μm).

Article Snippet: 5xFAD animals (C57BL/6J background, stock no: 034848) were purchased from the Jackson Laboratory and maintained in temperature-, light-, and humidity-controlled conditions with ad libitum access to food and water.

Techniques: Expressing, Gene Expression, MANN-WHITNEY, Staining

Journal: Neurotherapeutics

Article Title: Microglial GM3 accumulation impairs Aβ phagocytic activity and promotes neuroinflammation in Alzheimer's disease

doi: 10.1016/j.neurot.2026.e00851

Figure Lengend Snippet: Microglial Hexa and Hexb knockdown improves cognition and reduces both Aβ burden and neuroinflammation in 5xFAD mice. (A) Experimental scheme describing the timeline of virus delivery and subsequent behavioral testing. (B) In the familiarization session, all groups spent comparable amounts of time exploring the two identical objects. In the test session, scramble shRNA–treated 5xFAD mice displayed a reduced discrimination index compared to wild-type controls, whereas Hexa / Hexb shRNA–treated 5xFAD mice showed a significant rescue of cognitive deficits (∗∗p < 0.01, one-way ANOVA and Tukey's post hoc analysis, n = 4–6 per group). (C) Representative images of Aβ plaque deposition (left), along with quantitative analysis (right), show that Hexa / Hexb shRNA-injected 5xFAD mice display a reduced Aβ burden compared with scramble shRNA-treated 5xFAD mice (∗p < 0.05, ∗∗∗∗p < 0.0001, one-way ANOVA and Tukey's post hoc analysis, n = 3–4 per group, scale bar = 500 μm). (D) Representative Iba1 staining images (left) and quantitative assessment (right) indicate that Hexa / Hexb shRNA-injected 5xFAD mice show a substantial reduction in microglial activation compared with scramble shRNA-treated 5xFAD mice (∗∗p < 0.01, ∗∗∗∗p < 0.0001, one-way ANOVA and Tukey's post hoc analysis, n = 3–4 per group, scale bar = 500 μm). (E) Representative images of GFAP immunostaining (left) and corresponding quantification (right) reveal that Hexa / Hexb shRNA-treated 5xFAD mice exhibit a significantly attenuated astrocyte activation relative to scramble shRNA-injected 5xFAD mice (∗p < 0.05, ∗∗∗∗p < 0.0001, one-way ANOVA and Tukey's post hoc analysis, n = 3–4 per group, scale bar = 500 μm).

Article Snippet: 5xFAD animals (C57BL/6J background, stock no: 034848) were purchased from the Jackson Laboratory and maintained in temperature-, light-, and humidity-controlled conditions with ad libitum access to food and water.

Techniques: Knockdown, Virus, shRNA, Injection, Staining, Activation Assay, Immunostaining

Journal: Neurotherapeutics

Article Title: Microglial GM3 accumulation impairs Aβ phagocytic activity and promotes neuroinflammation in Alzheimer's disease

doi: 10.1016/j.neurot.2026.e00851

Figure Lengend Snippet: Schematic illustration of the proposed mechanism by which HexA contributes to AD pathology and how its inhibition may offer therapeutic benefits. Top panel: In 5xFAD mice, microglial overexpression of HexA (composed of α and β subunits) promotes lysosomal conversion of GM2 to GM3, leading to GM3 accumulation. Elevated GM3 impairs microglial Aβ phagocytosis and enhances neuroinflammatory responses. Bottom panel: Genetic inhibition of HexA by reducing Hexa and Hexb expression reduces GM3 accumulation, restores Aβ phagocytic capacity, and attenuates microglia-mediated inflammation, collectively contributing to improved AD pathology.

Article Snippet: 5xFAD animals (C57BL/6J background, stock no: 034848) were purchased from the Jackson Laboratory and maintained in temperature-, light-, and humidity-controlled conditions with ad libitum access to food and water.

Techniques: Inhibition, Over Expression, Expressing