Commensal microbiota divergently affect myeloid subsets in the mammalian central nervous system during homeostasis and disease (2021)
Figure 7

Figure 7: Increased Aβ uptake by perivascular macrophages in GF 5xFAD mice.

(A-C) Representative immunofluorescence images visualizing (A) cortical mMΦ, (B) cpMΦ located in the lateral ventricle and (C) hippocampal pvMΦ from SPF (left), GF (middle) and ABX-treated 5xFAD mice (right). Iba1 (green) and Aβ (6E10, red). Nuclei were stained with DAPI (blue). White arrowheads indicate Iba1+Aβ+ MΦ and white arrows show Iba1+Aβ- MΦ. Scale bar: 50 μm.

(D) Quantification of the number of 6E10+ Aβ-plaques per mm2. Each symbol represents one mouse. Means ± s.e.m. are indicated. Significant differences were evaluated by one-way ANOVA followed by Tukey's post-hoc comparison test and marked with asterisks (***P < 0.001). Data are representative of four independent experiments.

(E-G) Quantification of Iba1+ (E) mMΦ, (F) cpMΦ and (G) pvMΦ per mm2. Each symbol represents one mouse. At least three slides were examined per individual mouse. Data are presented as mean ± s.e.m. Significant differences were determined by two-way ANOVA followed by Tukey's post-hoc comparison test and marked with asterisks (*P < 0.05, **P < 0.01, ***P < 0.001). Data are representative of two independent experiments.

(H-J) Percentage of Iba1+Aβ+ (H) cortical mMΦ, (I) cpMΦ and (J) hippocampal pvMΦ of SPF, GF and ABX-treated 5xFAD mice. Means ± s.e.m. are indicated. Significant differences were evaluated by one-way ANOVA followed by Tukey's post-hoc comparison test and marked with asterisks (*P < 0.05). Data are representative of two independent experiments.

(K) Representative cytometric graph of methoxy-X-O4+ labelled pvMΦ from SPF (green line), GF (yellow line) and ABX-treated (blue line) 5xFAD mice and respective WT controls (dashed lines). Quantification of percentages of methoxy-X-O4+ labelled pvMΦ are depicted. Each symbol represents one mouse. Data are presented as mean ± s.e.m. Significant differences were determined by two-way ANOVA followed by Tukey's post-hoc comparison test (*P < 0.05, ***P < 0.001). Data are representative of three independent experiments.

TABLES

Cell counts per condition for all cells in Figure 1.

Cell counts per condition for microglia in Figure 2.

Cell counts per condition for cpMΦ in Figure 3.

Cell counts per condition for pvMΦ in Figure 4.

Cell counts per condition for mMΦ in Figure 5.

DATASETS

Cluster marker genes for all cells.

Hypergeometric test statistics for a) the enrichment of different compartments and b) GF and SPF cells in the clusters of all cells.

Genes along the microglia trajectory.

Genes along the CAM trajectory.

Overview of enriched GO terms in all cells in Appendix S1.

Cluster marker genes for microglia.

Hypergeometric test statistics for the enrichment of GF and SPF cells in the microglia clusters.

Results of differential gene expression analysis between SPF and GF microglia.

Cluster marker genes for cpMΦ.

Hypergeometric test statistics for the enrichment of GF and SPF cells in the cpMΦ clusters.

Results of differential gene expression analysis between SPF and GF a) CPepi and b) cpMΦ.

Cluster marker genes for pvMΦ.

Hypergeometric test statistics for the enrichment of GF and SPF cells in the pvMΦ clusters.

Results of differential gene expression analysis between SPF and GF pvMΦ.

Cluster marker genes for mMΦ.

Hypergeometric test statistics for the enrichment of GF and SPF cells in the mMΦ clusters.

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