Journal: Alzheimer's & Dementia

Article Title: Neuronal ABCA7 deficiency aggravates mitochondrial dysfunction and neurodegeneration in Alzheimer's disease

doi: 10.1002/alz.70112

Figure Lengend Snippet: Neuronal ABCA7 deficiency exacerbates neuronal damage in 10‐ to 12‐month‐old 5×FAD mice. (A–C) Quantification of X34‐positive or LAMP1‐positive area and total X34‐associated LAMP1 area in the cortex (B) and hippocampus (C) of 5×FAD; control (male, N = 2 to 3; female, N = 3) and 5×FAD; n Abca7 −/− mice (male, N = 3; female, N = 3). Scale bars, 50 µm. (D and E) Quantification of PSD95 and SYP by Western blotting in t cortex from 5×FAD control (male, N = 3; female, N = 3) and 5×FAD; n Abca7 −/− mice (male, N = 3; female, N = 3). (F) NfL levels measured by Quanterix Simoa NfL assay kit in plasma from 5×FAD control (male, N = 16; female, N = 13) and 5×FAD; n Abca7 −/− mice (male, N = 12; female, N = 9). Data are shown in mean ± SEM. Open and closed symbols indicate female and male mice, respectively. * p <0.05, and n.s. by two‐way ANOVA test after adjusting for sex. NfL, neurofilament light; n.s., not significant; SYP, synaptophysin.

Article Snippet: Plaque‐associated Iba1‐positive microglia, which surrounds X34‐positive amyloid plaque within 15 µm, was quantified with Imaris using surface and plot.

Techniques: Control, Western Blot

Journal: Alzheimer's & Dementia

Article Title: PLCG2 modulates TREM2 expression and signaling in response to Alzheimer's disease pathology

doi: 10.1002/alz.70231

Figure Lengend Snippet: PLCG2 deficiency results in reduced TREM2 mRNA and protein expression in 5xFAD mice. A, Representative images of plaques (X34, blue), microglia (Iba1, green), and TREM2 (red) for qualitative assessment in the cortices of 7.5‐month‐old mice. Scale bar, 100 µm. Representative immunoblot (B) and quantification (C and D) of PLCG2 and TREM2 protein expression in the cortices of 7.5‐month‐old B6 WT , 5xFAD WT , 5xFAD Plcg2 ‐/− , and 5xFAD Trem2 ‐/− mice ( n = 4 per group; 2 male and 2 female mice; TREM2 and PLCG2 expression were normalized with β‐Actin). PLCG2 (E) and TREM2 (F) mRNA expression reported as transcripts/million (TPM) as measured by bulk‐RNA sequencing of the cortices of 7.5‐month‐old mice ( n = 6 per group; 3 males and 3 females for 5xFAD WT and 5xFAD Trem2 ‐/‐ and 4 males and 2 females for 5xFAD Plcg2 ‐/− ). Male mice are marked with a square (□), and female mice are marked with a circle (°). All data are presented as the mean ± standard error of the mean analyzed by analysis of variance followed by Tukey multiple comparisons test. * p < 0.05; ** p < 0.01; *** p < 0.001. ns, not significant; PLCG2, phospholipase C gamma 2; TREM2, triggering receptor expressed on myeloid cells 2.

Article Snippet: After drying, the slides were stained with X34 (Millipore Sigma, SML1954) by first rehydrating in PBST and then incubating with X34 (100 μM) in Coplin jars for 10 minutes at RT.

Techniques: Expressing, Western Blot, RNA Sequencing

Journal: Alzheimer's & Dementia

Article Title: PLCG2 modulates TREM2 expression and signaling in response to Alzheimer's disease pathology

doi: 10.1002/alz.70231

Figure Lengend Snippet: PLCG2 and TREM2 deficiency exacerbate amyloid pathology and impair microglial engagement with Aβ plaques. (A) Representative images of dense‐core plaques (X34, blue) and dystrophic neurites (LAMP1, magenta) in the cortices of 7.5‐month‐old 5xFAD WT , 5xFAD Plcg2 ‐/− , and 5xFAD Trem2 ‐/− mice. Scale bar, 1000 µm. Percentage of X34+ area (B) or LAMP1+ area (D) in the cortices of 7.5‐month‐old mice of each genotype. Average X34+ plaque size (C), and LAMP1 total area per X34+ plaque (E) in the cortices of 7.5‐month‐old mice of each genotype. F, Representative images of dense‐core plaques (X34, blue) and microglia (Iba1, green) in cortices of 7.5‐month‐old mice. Total percentage of Iba1+ area (G) within the cortex, total percent of X34+ plaque area colocalized with Iba1+ area (H), and percentage of periplaque Iba1+ microglia (I) within a circular area with a radius of 25 µm centered on X34 + plaques ( n = 6 mice and > 100 plaques per genotype) in the cortices of 7.5‐month‐old mice of each genotype. J,K, Representative images illustrating periplaque regions in 5xFAD WT and 5xFAD Plcg2 ‐/− mice. Scale bar, 25 µm. n = 6 per group; 3 males and 3 females for 5xFAD WT and 5xFAD Trem2 ‐/‐ and 4 males and 2 females for 5xFAD Plcg2 ‐/− . Male mice are marked with a square (□), and female mice are marked with a circle (°). Data points represent means of three matched sagittal sections per mouse. All data are presented as the mean ± standard error of the mean analyzed by analysis of variance followed by Tukey multiple comparisons test. * p < 0.05; ** p < 0.01; *** p < 0.001. Aβ, amyloid beta; LAMP1, lysosomal‐associated membrane protein 1; ns, not significant; PLCG2, phospholipase C gamma; TREM2, triggering receptor expressed on myeloid cells 2.

Article Snippet: After drying, the slides were stained with X34 (Millipore Sigma, SML1954) by first rehydrating in PBST and then incubating with X34 (100 μM) in Coplin jars for 10 minutes at RT.

Techniques: Membrane

Journal: Alzheimer's & Dementia

Article Title: PLCG2 modulates TREM2 expression and signaling in response to Alzheimer's disease pathology

doi: 10.1002/alz.70231

Figure Lengend Snippet: PLCG2 and TREM2 deficiency similarly impair immune pathways. Cortices of 5xFAD WT , 5xFAD Plcg2 ‐/− , and 5xFAD Trem2 ‐/‐ mice were harvested at 7.5 months for bulk RNA‐sequencing analysis. A, Volcano plot showing significant DEGs (FDR < 0.05, FC > 1.5) between 5xFAD Plcg2 ‐/‐ and 5xFAD WT mice. B, Volcano plot showing significant DEGs (FDR < 0.05, FC > 1.5) between 5xFAD Trem2 ‐/‐ and 5xFAD WT . C, Top 10 GO terms for biological processes using all DEGs that met the significance and FC threshold between 5xFAD Plcg2 ‐/‐ and 5xFAD WT mice. D, Top 10 GO terms for biological processes using all DEGs that met the significance and FC threshold between 5xFAD Trem2 ‐/− and 5xFAD WT mice. E, Heatmap depicting mRNA expression of disease‐associated microglia genes between 5xFAD WT , 5xFAD Plcg2 ‐/− , and 5xFAD Trem2 ‐/‐ mice. F, Representative images of dense‐core plaques (X34, blue), microglia, (Iba1, green), and phagolysosomes (CD68, red). Scale bar, 100 µm. G, Total percentage of CD68 + area. H, Total percentage of CD68 area normalized to total Iba1 + area. I, Percentage of periplaque CD68 + area within a circular area with a radius of 25 µm centered on X34 + plaques. J, CD68 mRNA expression reported as transcripts/million (TPM). Representative immunoblot (K) and quantification (L) of CD11c protein from in 5xFAD WT , 5xFAD Plcg2 ‐/− , and 5xFAD Trem2 ‐/− cortices. Male mice are marked with a square (□), and female mice are marked with a circle (°). All data are presented as the mean ± standard error of the mean analyzed by analysis of variance followed by Tukey multiple comparisons test. * p < 0.05; ** p < 0.01; *** p < 0.001. DEGs, differentially expressed genes; FC, fold change; FDR, false discovery rate; GO, Gene Ontology; ns, not significant; PLCG2, phospholipase C gamma; TREM2, triggering receptor expressed on myeloid cells 2.

Article Snippet: After drying, the slides were stained with X34 (Millipore Sigma, SML1954) by first rehydrating in PBST and then incubating with X34 (100 μM) in Coplin jars for 10 minutes at RT.

Techniques: RNA Sequencing, Expressing, Western Blot