Journal: Alzheimer's Research & Therapy

Article Title: Microglia prevent beta-amyloid plaque formation in the early stage of an Alzheimer’s disease mouse model with suppression of glymphatic clearance

doi: 10.1186/s13195-020-00688-1

Figure Lengend Snippet: AQP4 deletion increased intraneuronal Aβ and IDE activity without causing plaque formation in the cerebral cortex of 3-month-old APP/PS1 mice. a , b Brain sections stained by thioflavine-S florescence and Aβ 1–40 immunohistochemistry showing no Aβ plaque disposition. c Double immunofluorescence for total-Aβ and NeuN. There was increased Aβ immunostaining in cerebral neurons of AQP4 −/− APP/PS1 mice compared to those in APP/PS1 controls. d , g Representative Western blot bands and densitometry analysis of APP, CTF-β, and Aβ monomer/oligomers. e , h Representative Western blot bands and densitometry analysis of BACE1, PS1, LRP1, IDE, and NEP in the cortex. f Quantification of Aβ-positive area fraction. i Enzyme activity assay of NEP and IDE. Data in f were analyzed by Student’s t test; other data were analyzed by the two-way ANOVA with Tukey’s post hoc test. Data are mean ± SEM. n = 4 per group. * p < 0.05; ** p < 0.01; *** p < 0.001

Article Snippet: Brain slices were blocked for 1 h at room temperature with 5% BSA, incubated with primary antibodies including rabbit anti-NeuN (1:500; Abcam, #ab177487), mouse anti-glial fibrillary acidic protein (GFAP) (1:800; Millipore, #AB3594), rabbit anti-GFAP (1:400; Abcam, #ab7260), rabbit anti-ionized calcium-binding adaptor molecule 1 (Iba1) (1:1000; Wako, #019-19741), rat anti-CD68 (1:100; bio-rad, #MCA1957), goat anti-lysosome-associated membrane glycoprotein 1 (Lamp1) (1:200, R&D systems, #AF4320), mouse anti-glutamine synthetase (GS) (1:100, BD, #610517), rabbit anti-Aβ 1–40 (1:250; CST, #12990), mouse anti-total Aβ (Aβ 1–16 ) (1:1000; Covance, # 803001), rabbit anti-AQP4 (1:400; Millipore, #AB3594), rabbit anti-apoE (1:200; Abcam, #ab20874), or mouse anti-apoE (1:200; Abcam, #ab1906) overnight at 4 °C.

Techniques: Activity Assay, Staining, Immunohistochemistry, Immunofluorescence, Immunostaining, Western Blot, Enzyme Activity Assay

Journal: Alzheimer's Research & Therapy

Article Title: Microglia prevent beta-amyloid plaque formation in the early stage of an Alzheimer’s disease mouse model with suppression of glymphatic clearance

doi: 10.1186/s13195-020-00688-1

Figure Lengend Snippet: Increased microglial activation, and Aβ plaque deposition following eliminating microglia in AQP4 −/− /APP/PS1 mice. a Double immunofluorescence for total-Aβ and Iba1. Microglia were apparently activated in the cerebral cortex of AQP4 −/− /APP/PS1 mice, compared to those in APP/PS1 controls. Dot-like signals of total-Aβ (arrowheads) were frequently observed in AQP4 −/− /APP/PS1 microglial cells. b , c Quantification of Iba1-positive, and iba1 and total-Aβ double-positive area fraction in the cerebral cortex, respectively. d , f Double immunofluorescence and quantification for CD68 and Iba1. CD68-positive microglia (arrowheads) were apparently increased in the cerebral cortex of AQP4 −/− /APP/PS1 mice, compared to those in APP/PS1 controls. e , g Double immunofluorescence and quantification for Lamp1 and Iba1. Lamp1-positive microglia (arrowheads) were also apparently increased in the cerebral cortex of AQP4 −/− /APP/PS1 mice. h , i Triple immunofluorescence and quantification for Lamp1, total-Aβ, and Iba1. Aβ immunoreactive products were restrictively localized to Lamp1-positive lysosome of Iba1-positive microglia. j Iba1 positive microglia were almost eliminated in both APP/PS1 mice and AQP4 −/− /APP/PS1 with clodronate liposome treatment. k , l Brain sections stained by thioflavine-S and Aβ 1–40 , respectively. Aβ plaque disposition was present at the cerebral cortex of AQP4 −/− /APP/PS1 mice received local injection of clodronate liposomes. m Double immunofluorescence for total-Aβ and NeuN. Total-Aβ immunoreactivity (arrowheads) was increased within the cytoplasm of NeuN positive neurons of AQP4 −/− /APP/PS1 mice injected clodronate liposomes. n – s Quantification of Iba1, thioflavine-S, Aβ 1–40 , and total-Aβ-positive area fraction or number in the cerebral cortex, respectively. Data in c , f , g , and i were analyzed by Student’s t test and in b , n , q , and s were analyzed by the two-way ANOVA with Newman-Keuls post hoc test. Data are mean ± SEM, n = 4 per group, * p < 0.05; ** p < 0.01; *** p < 0.001

Article Snippet: Brain slices were blocked for 1 h at room temperature with 5% BSA, incubated with primary antibodies including rabbit anti-NeuN (1:500; Abcam, #ab177487), mouse anti-glial fibrillary acidic protein (GFAP) (1:800; Millipore, #AB3594), rabbit anti-GFAP (1:400; Abcam, #ab7260), rabbit anti-ionized calcium-binding adaptor molecule 1 (Iba1) (1:1000; Wako, #019-19741), rat anti-CD68 (1:100; bio-rad, #MCA1957), goat anti-lysosome-associated membrane glycoprotein 1 (Lamp1) (1:200, R&D systems, #AF4320), mouse anti-glutamine synthetase (GS) (1:100, BD, #610517), rabbit anti-Aβ 1–40 (1:250; CST, #12990), mouse anti-total Aβ (Aβ 1–16 ) (1:1000; Covance, # 803001), rabbit anti-AQP4 (1:400; Millipore, #AB3594), rabbit anti-apoE (1:200; Abcam, #ab20874), or mouse anti-apoE (1:200; Abcam, #ab1906) overnight at 4 °C.

Techniques: Activation Assay, Immunofluorescence, Staining, Injection