gfp htau plasmids  (Addgene inc)

Journal: Zoological Research

Article Title: Novel mouse model of Alzheimer’s disease exhibits pathology through synergistic interactions among amyloid-β, tau, and reactive astrogliosis

doi: 10.24272/j.issn.2095-8137.2024.257

Figure Lengend Snippet: TauP301L overexpression and adenovirus-induced reactive astrogliosis in the hippocampus of APP/PS1 mice induces phosphorylated tau, reactive astrogliosis, and NFTs A: Representative immunohistochemical images of S199 (p-tau, red) and GFAP (astrocyte, blue) in 8-month-old WT-Control, APP/PS1-Control, APP/PS1-TauP301L, and APP/PS1-TauP301L-Adeno mice. Scale bars: 40 μm. B, C: Bar graph showing mean intensity of GFAP (B) and S199 (C). Each bar represents mean±SEM. Sample sizes: WT-control ( n =10), APP/PS1-Control ( n =9), APP/PS1-TauP301L ( n =11), APP/PS1-TauP301L-Adeno ( n =14), n : number of slices, collected from 3–4 mice per group. Mean±SEM values: (B) WT-control=439.354±36.525, APP/PS1-Control=1 007.23±69.796, APP/PS1-TauP301L=1 489.587±61.307, APP/PS1-TauP301L-Adeno=2 137.64±123.507. (C) WT-control=11.963±2.66, APP/PS1-Control=13.645±1.712, APP/PS1-TauP301L=96.347±9.63, and APP/PS1-TauP301L-Adeno=227.446±32.95. D: Representative Sholl analysis images of GFAP-positive astrocytes in WT-Control, APP/PS1-Control, APP/PS1-TauP301L, and APP/PS1-TauP301L-Adeno mice. Circles centered around soma are separated by radial intervals of 10 μm. Scale bars: 50 μm. E: Bar graph showing number of intersections in GFAP-positive cells. Each bar represents mean±SEM. Sample sizes: WT-control ( n =9), APP/PS1-Control ( n =13), APP/PS1-TauP301L ( n =13), APP/PS1-TauP301L-Adeno ( n =9), n : GFAP-positive cells collected from 3–4 mice per group. Mean±SEM values: WT-control=60.333±7.878, APP/PS1-Control=91.538±3.847, APP/PS1-TauP301L=82.069±4.423, and APP/PS1-TauP301L-Adeno=124.666±13.497. F: Representative photomicrographs of NFTs stained using ABC/DAB in APP/PS1-TauP301L-Adeno brain sections. Scale bars: 500 μm. G, H: Bar graph showing quantification of NFT-positive intensity per area (100 μm 2 ) in CA1 and DG of hippocampus. Each bar represents mean±SEM. (G, CA1 pyramidal layer). Sample sizes: WT-Control ( n =23), WT-TauP301L/Adeno ( n =24), APP/PS1-TauP301L ( n =27), and APP/PS1-TauP301L-Adeno ( n =23). (H, DG granule cell layer) Sample sizes: WT-Control ( n =22), WT-TauP301L/Adeno ( n =22), APP/PS1-TauP301L ( n =31), and APP/PS1-TauP301L-Adeno ( n =33). Each dot represents an individual area measurement, with data collected from 3–6 mice per group. Mean±SEM values: (G, CA1 pyramidal layer) WT-Control=1.459±0.588, WT-TauP301L/Adeno=5.382±1.043, APP/PS1-TauP301L=5.647±0.804, APP/PS1-TauP301L-Adeno=9.199±0.576, (H, DG granule cell layer) WT-Control=2.604±0.662, WT-TauP301L/Adeno=11.88±0.964, APP/PS1-TauP301L=15.006±0.825, and APP/PS1-TauP301L-Adeno=28.19±0.832. Statistical analysis was conducted using one-way ANOVA with Tukey’s post-hoc test, ns: Not significant; * : P <0.05; ** : P <0.01; *** : P <0.001; **** : P <0.0001.

Article Snippet: For AAV-based TauP301L expression, the TauP301L sequence from pRK5-EGFP-TauP301L (Addgene, #46908, USA) was inserted into the AAV-EF1α-EGFP vector backbone (Addgene, #60058, USA) at the KpnI and NcoI restriction sites using infusion cloning (Clontech, #639649, USA).

Techniques: Over Expression, Immunohistochemical staining, Control, Staining

Journal: Zoological Research

Article Title: Novel mouse model of Alzheimer’s disease exhibits pathology through synergistic interactions among amyloid-β, tau, and reactive astrogliosis

doi: 10.24272/j.issn.2095-8137.2024.257

Figure Lengend Snippet: TauP301L overexpression and adenovirus-induced reactive astrogliosis in the hippocampus of APP/PS1 mice induces robust amyloid pathologies A: Representative immunofluorescence images of Aβ (magenta) staining in hippocampal CA1 from WT-Control, APP/PS1-Control, APP/PS1-TauP301L, and APP/PS1-TauP301L-Adeno mice. Scale bars: 40 μm. B: Bar graph showing quantification of Aβ plaque size in the hippocampus. Each bar represents mean±SEM. Sample sizes: WT-control ( n =8), APP/PS1-Control ( n =13), APP/PS1-TauP301L ( n =17), APP/PS1-TauP301L-Adeno ( n =18), n : number of slices, collected from three mice per group. Mean±SEM values: WT-control=1.924±1.329, APP/PS1-Control=40.663±8.291, APP/PS1-TauP301L=91.938±10.201, and APP/PS1-TauP301L-Adeno=299.923±57.433. C: Representative images showing Aβ plaques stained with thioflavin-S (thio-S). S1 indicates S1 somatosensory cortex, Pir indicates piriform cortex. Scale bars: 1 mm. D, E: Bar graph showing quantification of number of Aβ plaques in the S1 somatosensory cortex (D) and piriform cortex (E). Each bar represents mean±SEM. Sample sizes: (D, E) WT-control ( n =10), APP/PS1-Control ( n =8), APP/PS1-TauP301L ( n =10), APP/PS1-TauP301L-Adeno ( n =8), with data collected from three mice per group. Mean±SEM values: (D) WT-control=0, APP/PS1-Control=158.038±24.752, APP/PS1-TauP301L=217.972±18.255, APP/PS1-TauP301L-Adeno=347.858±29.539. (E) WT-control=0, APP/PS1-Control=140.775±19.422, APP/PS1-TauP301L=224.287±18.873, APP/PS1-TauP301L-Adeno=402.746±37.664. Statistical analysis was conducted using one-way ANOVA with Tukey’s post-hoc test, * : P <0.05; *** : P <0.001; **** : P <0.0001.

Article Snippet: For AAV-based TauP301L expression, the TauP301L sequence from pRK5-EGFP-TauP301L (Addgene, #46908, USA) was inserted into the AAV-EF1α-EGFP vector backbone (Addgene, #60058, USA) at the KpnI and NcoI restriction sites using infusion cloning (Clontech, #639649, USA).

Techniques: Over Expression, Immunofluorescence, Staining, Control

Journal: Zoological Research

Article Title: Novel mouse model of Alzheimer’s disease exhibits pathology through synergistic interactions among amyloid-β, tau, and reactive astrogliosis

doi: 10.24272/j.issn.2095-8137.2024.257

Figure Lengend Snippet: TauP301L overexpression and adenovirus-induced reactive astrogliosis in the hippocampus of APP/PS1 mice increases astrocytic GABA and MAO-B enzyme activity A: Representative confocal images of astrocytes (GFAP, green) and GABA (red) in the molecular layer of CA3 from 8-month-old WT-Control, APP/PS1-Control, APP/PS1-TauP301L, and APP/PS1-TauP301L-Adeno mice. Scale bars: 5 μm. B: Violin plots showing GABA intensity in GFAP-positive astrocytes, representing the distribution of data within each group. Dashed lines indicate median, dotted lines represent quartiles. Sample sizes: WT-Control ( n =28), APP/PS1-Control ( n =38), APP/PS1-TauP301L ( n =31), APP/PS1-TauP301L/Adeno ( n =41), with GFAP-positive cells collected from four mice per group. Mean±SEM values: WT-Control=31.931±13.091, APP/PS1-Control=61.623±15.829, APP/PS1-TauP301L=200.939±33.855, and APP/PS1-TauP301L-Adeno=274.775±23.761. C: Schematic of MAO-B enzyme activity assay in the hippocampus (Top). Representative images of color change after 20 min of MAO-B reaction (Bottom, n =2 for 8-month-old WT-Control; n =3 for other groups). D: Bar graph showing MAO-B enzyme activity measured by absorbance at 570 nm in hippocampal tissue. Each bar represents mean absorbance±SEM, with sample sizes: WT-Control ( n =5), APP/PS1-Control ( n =6), APP/PS1-TauP301L ( n =3), and APP/PS1-TauP301L-Adeno ( n =3). One hippocampal tissue sample from one hemisphere was examined per mouse. Mean absorbance±SEM values: non-treated WT-control=0.157±0.004, selegiline-treated WT-control=0.052±0.003; non-treated APP/PS1-Control=0.211±0.007, selegiline-treated APP/PS1-Control=0.058±0.003; on-treated APP/PS1-TauP301L=0.22±0.015, selegiline-treated APP/PS1-TauP301L=0.06±0.004; non-treated APP/PS1-TauP301L-Adeno=0.24±0.023, and selegiline-treated APP/PS1-TauP301L-Adeno=0.24±0.023. Statistical analysis was conducted using one-way ANOVA with Tukey’s post-hoc test, *** : P <0.001; **** : P <0.0001.

Article Snippet: For AAV-based TauP301L expression, the TauP301L sequence from pRK5-EGFP-TauP301L (Addgene, #46908, USA) was inserted into the AAV-EF1α-EGFP vector backbone (Addgene, #60058, USA) at the KpnI and NcoI restriction sites using infusion cloning (Clontech, #639649, USA).

Techniques: Over Expression, Activity Assay, Control, Enzyme Activity Assay

Journal: Zoological Research

Article Title: Novel mouse model of Alzheimer’s disease exhibits pathology through synergistic interactions among amyloid-β, tau, and reactive astrogliosis

doi: 10.24272/j.issn.2095-8137.2024.257

Figure Lengend Snippet: TauP301L overexpression and adenovirus-induced reactive astrogliosis in the hippocampus of APP/PS1 mice accelerates neurodegeneration A: Representative confocal images of NeuN (neuron, white) in the CA1 pyramidal layer of 8-month-old WT-Control, APP/PS1-Control, APP/PS1-TauP301L, and APP/PS1-TauP301L-Adeno mice. Scale bars: 40 μm. B: Bar graph showing number of NeuN-positive cells per area (100 µm 2 ). Each bar represents mean±SEM. Sample sizes for all groups: n =10, with each dot representing the average count from three area per slice. Ten slices were analyzed from 3–4 mice per group. Mean±SEM values: WT-Control=2.605±0.069, APP/PS1-Control=2.995±0.157, APP/PS1-TauP301L=2.593±0.091, and APP/PS1-TauP301L-Adeno=0.895±0.226). Statistical analysis was conducted using one-way ANOVA with Tukey’s post-hoc test, **** : P <0.0001.

Article Snippet: For AAV-based TauP301L expression, the TauP301L sequence from pRK5-EGFP-TauP301L (Addgene, #46908, USA) was inserted into the AAV-EF1α-EGFP vector backbone (Addgene, #60058, USA) at the KpnI and NcoI restriction sites using infusion cloning (Clontech, #639649, USA).

Techniques: Over Expression, Control

Journal: Zoological Research

Article Title: Novel mouse model of Alzheimer’s disease exhibits pathology through synergistic interactions among amyloid-β, tau, and reactive astrogliosis

doi: 10.24272/j.issn.2095-8137.2024.257

Figure Lengend Snippet: TauP301L overexpression and adenovirus-induced reactive astrogliosis in the hippocampus of APP/PS1 mice impairs hippocampus-dependent memory A, B: Schematic of novel object recognition (NOR) test (A) and representative heatmap image in NOR square chamber (gray) during test session (B). C: Bar graph showing discrimination index, calculated as: Discrimination index=exploration time in novel object/total exploration time (novel+familiar). Each bar represents mean±SEM, with sample sizes (number of mice): WT-Control ( n =16), WT-TauP301L ( n =5), WT-TauP301L/Adeno ( n =5), APP/PS1-Control ( n =10), APP/PS1-TauP301L ( n =13), and APP/PS1-TauP301L-Adeno ( n =19). Statistical analysis was conducted using two-way ANOVA with Bonferroni test, ns: Not significant; * : P <0.05; ** : P <0.01; **** : P <0.0001. Mean±SEM values: (Familiarization) WT-Control=0.475±0.016, WT-TauP301L=0.475±0.019, WT-TauP301L/Adeno=0.499±0.012, APP/PS1-Control=0.476±0.009, APP/PS1-TauP301L=0.497±0.019 and APP/PS1-TauP301L-Adeno=0.507±0.019. (Test) WT-Control=0.65±0.026, WT-TauP301L=0.56±0.025, WT-TauP301L/Adeno=0.7±0.037, APP/PS1-Control=0.602±0.039, APP/PS1-TauP301L=0.603±0.031, and APP/PS1-TauP301L-Adeno=0.513±0.028. D, E: Schematic of Y-maze (D) and representative heatmap of tracks (E) in Y-maze spontaneous alternation test. F: Bar graph showing average percentage of triad alternations relative to total arm entries. Each bar represents mean±SEM, with sample sizes (number of mice): WT-Control ( n =6), APP/PS1-Control ( n =7), APP/PS1-TauP301L ( n =8), and APP/PS1-TauP301L-Adeno ( n =7). Statistical analysis was conducted using one-way ANOVA with Tukey’s post-hoc test, * : P <0.05; ** : P <0.01. Mean±SEM values: WT-Control=65.855±2.892, APP/PS1-Control=59.946±2.999, APP/PS1-TauP301L=68.399±2.794, and APP/PS1-TauP301L-Adeno=52.629±3.402. G: Schematic of passive avoidance test (PAT). H: Bar graph showing time for latency to enter dark chamber. Each bar represents mean±SEM, with sample sizes (number of mice): WT-Control ( n =6), WT-TauP301L ( n =5), WT-TauP301L-Adeno ( n =5), APP/PS1-Control ( n =7), APP/PS1-TauP301L ( n =8), and APP/PS1-TauP301L-Adeno ( n =7). Statistical analysis was conducted using two-way ANOVA with Bonferroni test, ns: Not significant; ** : P <0.01. Mean±SEM values: (Acquisition) WT-Control=20.75±3.748, WT-TauP301L=29.8±10.918, WT-TauP301L-Adeno=32.9±8.532, APP/PS1-Control=38.214±12.636, APP/PS1-TauP301L=29.188±6.408, APP/PS1-TauP301L-Adeno=63.886±21.87. (Retention) WT-Control=421.85±80.612, WT-TauP301L=322.5±101.528, WT-TauP301L-Adeno=342.4±119.582, APP/PS1-Control=203.614±53.403, APP/PS1-TauP301L=319.55±43.637, and APP/PS1-TauP301L-Adeno=142.471±58.3.

Article Snippet: For AAV-based TauP301L expression, the TauP301L sequence from pRK5-EGFP-TauP301L (Addgene, #46908, USA) was inserted into the AAV-EF1α-EGFP vector backbone (Addgene, #60058, USA) at the KpnI and NcoI restriction sites using infusion cloning (Clontech, #639649, USA).

Techniques: Over Expression, Control

Journal: Zoological Research

Article Title: Novel mouse model of Alzheimer’s disease exhibits pathology through synergistic interactions among amyloid-β, tau, and reactive astrogliosis

doi: 10.24272/j.issn.2095-8137.2024.257

Figure Lengend Snippet: Summary of APP/PS1-TauP301L-Adeno mouse model Summary figure illustrates progression of pathological features in both APP/PS1-TauP301L and APP/PS1-TauP301L-Adeno mice. At 8 months of age, 3 months post-injection, APP/PS1-TauP301L mice exhibited increased reactive astrogliosis and accumulation of Aβ plaque in the hippocampus compared to APP/PS1-control mice, but did not display any significant impairment of hippocampus-dependent cognitive function. In contrast, APP/PS1-TauP301L-Adeno mice showed exacerbated reactive astrogliosis and further accumulation of Aβ plaques, as well as NFT development, further exacerbating hippocampus-dependent cognitive impairment.

Article Snippet: For AAV-based TauP301L expression, the TauP301L sequence from pRK5-EGFP-TauP301L (Addgene, #46908, USA) was inserted into the AAV-EF1α-EGFP vector backbone (Addgene, #60058, USA) at the KpnI and NcoI restriction sites using infusion cloning (Clontech, #639649, USA).

Techniques: Injection, Control

Journal: Zoological Research

Article Title: Novel mouse model of Alzheimer’s disease exhibits pathology through synergistic interactions among amyloid-β, tau, and reactive astrogliosis

doi: 10.24272/j.issn.2095-8137.2024.257

Figure Lengend Snippet: TauP301L overexpression and adenovirus-induced reactive astrogliosis in the hippocampus of APP/PS1 mice induces phosphorylated tau, reactive astrogliosis, and NFTs A: Representative immunohistochemical images of S199 (p-tau, red) and GFAP (astrocyte, blue) in 8-month-old WT-Control, APP/PS1-Control, APP/PS1-TauP301L, and APP/PS1-TauP301L-Adeno mice. Scale bars: 40 μm. B, C: Bar graph showing mean intensity of GFAP (B) and S199 (C). Each bar represents mean±SEM. Sample sizes: WT-control ( n =10), APP/PS1-Control ( n =9), APP/PS1-TauP301L ( n =11), APP/PS1-TauP301L-Adeno ( n =14), n : number of slices, collected from 3–4 mice per group. Mean±SEM values: (B) WT-control=439.354±36.525, APP/PS1-Control=1 007.23±69.796, APP/PS1-TauP301L=1 489.587±61.307, APP/PS1-TauP301L-Adeno=2 137.64±123.507. (C) WT-control=11.963±2.66, APP/PS1-Control=13.645±1.712, APP/PS1-TauP301L=96.347±9.63, and APP/PS1-TauP301L-Adeno=227.446±32.95. D: Representative Sholl analysis images of GFAP-positive astrocytes in WT-Control, APP/PS1-Control, APP/PS1-TauP301L, and APP/PS1-TauP301L-Adeno mice. Circles centered around soma are separated by radial intervals of 10 μm. Scale bars: 50 μm. E: Bar graph showing number of intersections in GFAP-positive cells. Each bar represents mean±SEM. Sample sizes: WT-control ( n =9), APP/PS1-Control ( n =13), APP/PS1-TauP301L ( n =13), APP/PS1-TauP301L-Adeno ( n =9), n : GFAP-positive cells collected from 3–4 mice per group. Mean±SEM values: WT-control=60.333±7.878, APP/PS1-Control=91.538±3.847, APP/PS1-TauP301L=82.069±4.423, and APP/PS1-TauP301L-Adeno=124.666±13.497. F: Representative photomicrographs of NFTs stained using ABC/DAB in APP/PS1-TauP301L-Adeno brain sections. Scale bars: 500 μm. G, H: Bar graph showing quantification of NFT-positive intensity per area (100 μm 2 ) in CA1 and DG of hippocampus. Each bar represents mean±SEM. (G, CA1 pyramidal layer). Sample sizes: WT-Control ( n =23), WT-TauP301L/Adeno ( n =24), APP/PS1-TauP301L ( n =27), and APP/PS1-TauP301L-Adeno ( n =23). (H, DG granule cell layer) Sample sizes: WT-Control ( n =22), WT-TauP301L/Adeno ( n =22), APP/PS1-TauP301L ( n =31), and APP/PS1-TauP301L-Adeno ( n =33). Each dot represents an individual area measurement, with data collected from 3–6 mice per group. Mean±SEM values: (G, CA1 pyramidal layer) WT-Control=1.459±0.588, WT-TauP301L/Adeno=5.382±1.043, APP/PS1-TauP301L=5.647±0.804, APP/PS1-TauP301L-Adeno=9.199±0.576, (H, DG granule cell layer) WT-Control=2.604±0.662, WT-TauP301L/Adeno=11.88±0.964, APP/PS1-TauP301L=15.006±0.825, and APP/PS1-TauP301L-Adeno=28.19±0.832. Statistical analysis was conducted using one-way ANOVA with Tukey’s post-hoc test, ns: Not significant; * : P <0.05; ** : P <0.01; *** : P <0.001; **** : P <0.0001.

Article Snippet: For AAV-based TauP301L expression, the TauP301L sequence from pRK5-EGFP-TauP301L (Addgene, #46908, USA) was inserted into the AAV-EF1α-EGFP vector backbone (Addgene, #60058, USA) at the KpnI and NcoI restriction sites using infusion cloning (Clontech, #639649, USA).

Techniques: Over Expression, Immunohistochemical staining, Control, Staining

Journal: Zoological Research

Article Title: Novel mouse model of Alzheimer’s disease exhibits pathology through synergistic interactions among amyloid-β, tau, and reactive astrogliosis

doi: 10.24272/j.issn.2095-8137.2024.257

Figure Lengend Snippet: TauP301L overexpression and adenovirus-induced reactive astrogliosis in the hippocampus of APP/PS1 mice induces robust amyloid pathologies A: Representative immunofluorescence images of Aβ (magenta) staining in hippocampal CA1 from WT-Control, APP/PS1-Control, APP/PS1-TauP301L, and APP/PS1-TauP301L-Adeno mice. Scale bars: 40 μm. B: Bar graph showing quantification of Aβ plaque size in the hippocampus. Each bar represents mean±SEM. Sample sizes: WT-control ( n =8), APP/PS1-Control ( n =13), APP/PS1-TauP301L ( n =17), APP/PS1-TauP301L-Adeno ( n =18), n : number of slices, collected from three mice per group. Mean±SEM values: WT-control=1.924±1.329, APP/PS1-Control=40.663±8.291, APP/PS1-TauP301L=91.938±10.201, and APP/PS1-TauP301L-Adeno=299.923±57.433. C: Representative images showing Aβ plaques stained with thioflavin-S (thio-S). S1 indicates S1 somatosensory cortex, Pir indicates piriform cortex. Scale bars: 1 mm. D, E: Bar graph showing quantification of number of Aβ plaques in the S1 somatosensory cortex (D) and piriform cortex (E). Each bar represents mean±SEM. Sample sizes: (D, E) WT-control ( n =10), APP/PS1-Control ( n =8), APP/PS1-TauP301L ( n =10), APP/PS1-TauP301L-Adeno ( n =8), with data collected from three mice per group. Mean±SEM values: (D) WT-control=0, APP/PS1-Control=158.038±24.752, APP/PS1-TauP301L=217.972±18.255, APP/PS1-TauP301L-Adeno=347.858±29.539. (E) WT-control=0, APP/PS1-Control=140.775±19.422, APP/PS1-TauP301L=224.287±18.873, APP/PS1-TauP301L-Adeno=402.746±37.664. Statistical analysis was conducted using one-way ANOVA with Tukey’s post-hoc test, * : P <0.05; *** : P <0.001; **** : P <0.0001.

Article Snippet: For AAV-based TauP301L expression, the TauP301L sequence from pRK5-EGFP-TauP301L (Addgene, #46908, USA) was inserted into the AAV-EF1α-EGFP vector backbone (Addgene, #60058, USA) at the KpnI and NcoI restriction sites using infusion cloning (Clontech, #639649, USA).

Techniques: Over Expression, Immunofluorescence, Staining, Control

Journal: Zoological Research

Article Title: Novel mouse model of Alzheimer’s disease exhibits pathology through synergistic interactions among amyloid-β, tau, and reactive astrogliosis

doi: 10.24272/j.issn.2095-8137.2024.257

Figure Lengend Snippet: TauP301L overexpression and adenovirus-induced reactive astrogliosis in the hippocampus of APP/PS1 mice increases astrocytic GABA and MAO-B enzyme activity A: Representative confocal images of astrocytes (GFAP, green) and GABA (red) in the molecular layer of CA3 from 8-month-old WT-Control, APP/PS1-Control, APP/PS1-TauP301L, and APP/PS1-TauP301L-Adeno mice. Scale bars: 5 μm. B: Violin plots showing GABA intensity in GFAP-positive astrocytes, representing the distribution of data within each group. Dashed lines indicate median, dotted lines represent quartiles. Sample sizes: WT-Control ( n =28), APP/PS1-Control ( n =38), APP/PS1-TauP301L ( n =31), APP/PS1-TauP301L/Adeno ( n =41), with GFAP-positive cells collected from four mice per group. Mean±SEM values: WT-Control=31.931±13.091, APP/PS1-Control=61.623±15.829, APP/PS1-TauP301L=200.939±33.855, and APP/PS1-TauP301L-Adeno=274.775±23.761. C: Schematic of MAO-B enzyme activity assay in the hippocampus (Top). Representative images of color change after 20 min of MAO-B reaction (Bottom, n =2 for 8-month-old WT-Control; n =3 for other groups). D: Bar graph showing MAO-B enzyme activity measured by absorbance at 570 nm in hippocampal tissue. Each bar represents mean absorbance±SEM, with sample sizes: WT-Control ( n =5), APP/PS1-Control ( n =6), APP/PS1-TauP301L ( n =3), and APP/PS1-TauP301L-Adeno ( n =3). One hippocampal tissue sample from one hemisphere was examined per mouse. Mean absorbance±SEM values: non-treated WT-control=0.157±0.004, selegiline-treated WT-control=0.052±0.003; non-treated APP/PS1-Control=0.211±0.007, selegiline-treated APP/PS1-Control=0.058±0.003; on-treated APP/PS1-TauP301L=0.22±0.015, selegiline-treated APP/PS1-TauP301L=0.06±0.004; non-treated APP/PS1-TauP301L-Adeno=0.24±0.023, and selegiline-treated APP/PS1-TauP301L-Adeno=0.24±0.023. Statistical analysis was conducted using one-way ANOVA with Tukey’s post-hoc test, *** : P <0.001; **** : P <0.0001.

Article Snippet: For AAV-based TauP301L expression, the TauP301L sequence from pRK5-EGFP-TauP301L (Addgene, #46908, USA) was inserted into the AAV-EF1α-EGFP vector backbone (Addgene, #60058, USA) at the KpnI and NcoI restriction sites using infusion cloning (Clontech, #639649, USA).

Techniques: Over Expression, Activity Assay, Control, Enzyme Activity Assay

Journal: Zoological Research

Article Title: Novel mouse model of Alzheimer’s disease exhibits pathology through synergistic interactions among amyloid-β, tau, and reactive astrogliosis

doi: 10.24272/j.issn.2095-8137.2024.257

Figure Lengend Snippet: TauP301L overexpression and adenovirus-induced reactive astrogliosis in the hippocampus of APP/PS1 mice accelerates neurodegeneration A: Representative confocal images of NeuN (neuron, white) in the CA1 pyramidal layer of 8-month-old WT-Control, APP/PS1-Control, APP/PS1-TauP301L, and APP/PS1-TauP301L-Adeno mice. Scale bars: 40 μm. B: Bar graph showing number of NeuN-positive cells per area (100 µm 2 ). Each bar represents mean±SEM. Sample sizes for all groups: n =10, with each dot representing the average count from three area per slice. Ten slices were analyzed from 3–4 mice per group. Mean±SEM values: WT-Control=2.605±0.069, APP/PS1-Control=2.995±0.157, APP/PS1-TauP301L=2.593±0.091, and APP/PS1-TauP301L-Adeno=0.895±0.226). Statistical analysis was conducted using one-way ANOVA with Tukey’s post-hoc test, **** : P <0.0001.

Article Snippet: For AAV-based TauP301L expression, the TauP301L sequence from pRK5-EGFP-TauP301L (Addgene, #46908, USA) was inserted into the AAV-EF1α-EGFP vector backbone (Addgene, #60058, USA) at the KpnI and NcoI restriction sites using infusion cloning (Clontech, #639649, USA).

Techniques: Over Expression, Control

Journal: Zoological Research

Article Title: Novel mouse model of Alzheimer’s disease exhibits pathology through synergistic interactions among amyloid-β, tau, and reactive astrogliosis

doi: 10.24272/j.issn.2095-8137.2024.257

Figure Lengend Snippet: TauP301L overexpression and adenovirus-induced reactive astrogliosis in the hippocampus of APP/PS1 mice impairs hippocampus-dependent memory A, B: Schematic of novel object recognition (NOR) test (A) and representative heatmap image in NOR square chamber (gray) during test session (B). C: Bar graph showing discrimination index, calculated as: Discrimination index=exploration time in novel object/total exploration time (novel+familiar). Each bar represents mean±SEM, with sample sizes (number of mice): WT-Control ( n =16), WT-TauP301L ( n =5), WT-TauP301L/Adeno ( n =5), APP/PS1-Control ( n =10), APP/PS1-TauP301L ( n =13), and APP/PS1-TauP301L-Adeno ( n =19). Statistical analysis was conducted using two-way ANOVA with Bonferroni test, ns: Not significant; * : P <0.05; ** : P <0.01; **** : P <0.0001. Mean±SEM values: (Familiarization) WT-Control=0.475±0.016, WT-TauP301L=0.475±0.019, WT-TauP301L/Adeno=0.499±0.012, APP/PS1-Control=0.476±0.009, APP/PS1-TauP301L=0.497±0.019 and APP/PS1-TauP301L-Adeno=0.507±0.019. (Test) WT-Control=0.65±0.026, WT-TauP301L=0.56±0.025, WT-TauP301L/Adeno=0.7±0.037, APP/PS1-Control=0.602±0.039, APP/PS1-TauP301L=0.603±0.031, and APP/PS1-TauP301L-Adeno=0.513±0.028. D, E: Schematic of Y-maze (D) and representative heatmap of tracks (E) in Y-maze spontaneous alternation test. F: Bar graph showing average percentage of triad alternations relative to total arm entries. Each bar represents mean±SEM, with sample sizes (number of mice): WT-Control ( n =6), APP/PS1-Control ( n =7), APP/PS1-TauP301L ( n =8), and APP/PS1-TauP301L-Adeno ( n =7). Statistical analysis was conducted using one-way ANOVA with Tukey’s post-hoc test, * : P <0.05; ** : P <0.01. Mean±SEM values: WT-Control=65.855±2.892, APP/PS1-Control=59.946±2.999, APP/PS1-TauP301L=68.399±2.794, and APP/PS1-TauP301L-Adeno=52.629±3.402. G: Schematic of passive avoidance test (PAT). H: Bar graph showing time for latency to enter dark chamber. Each bar represents mean±SEM, with sample sizes (number of mice): WT-Control ( n =6), WT-TauP301L ( n =5), WT-TauP301L-Adeno ( n =5), APP/PS1-Control ( n =7), APP/PS1-TauP301L ( n =8), and APP/PS1-TauP301L-Adeno ( n =7). Statistical analysis was conducted using two-way ANOVA with Bonferroni test, ns: Not significant; ** : P <0.01. Mean±SEM values: (Acquisition) WT-Control=20.75±3.748, WT-TauP301L=29.8±10.918, WT-TauP301L-Adeno=32.9±8.532, APP/PS1-Control=38.214±12.636, APP/PS1-TauP301L=29.188±6.408, APP/PS1-TauP301L-Adeno=63.886±21.87. (Retention) WT-Control=421.85±80.612, WT-TauP301L=322.5±101.528, WT-TauP301L-Adeno=342.4±119.582, APP/PS1-Control=203.614±53.403, APP/PS1-TauP301L=319.55±43.637, and APP/PS1-TauP301L-Adeno=142.471±58.3.

Article Snippet: For AAV-based TauP301L expression, the TauP301L sequence from pRK5-EGFP-TauP301L (Addgene, #46908, USA) was inserted into the AAV-EF1α-EGFP vector backbone (Addgene, #60058, USA) at the KpnI and NcoI restriction sites using infusion cloning (Clontech, #639649, USA).

Techniques: Over Expression, Control

Journal: Zoological Research

Article Title: Novel mouse model of Alzheimer’s disease exhibits pathology through synergistic interactions among amyloid-β, tau, and reactive astrogliosis

doi: 10.24272/j.issn.2095-8137.2024.257

Figure Lengend Snippet: Summary of APP/PS1-TauP301L-Adeno mouse model Summary figure illustrates progression of pathological features in both APP/PS1-TauP301L and APP/PS1-TauP301L-Adeno mice. At 8 months of age, 3 months post-injection, APP/PS1-TauP301L mice exhibited increased reactive astrogliosis and accumulation of Aβ plaque in the hippocampus compared to APP/PS1-control mice, but did not display any significant impairment of hippocampus-dependent cognitive function. In contrast, APP/PS1-TauP301L-Adeno mice showed exacerbated reactive astrogliosis and further accumulation of Aβ plaques, as well as NFT development, further exacerbating hippocampus-dependent cognitive impairment.

Article Snippet: For AAV-based TauP301L expression, the TauP301L sequence from pRK5-EGFP-TauP301L (Addgene, #46908, USA) was inserted into the AAV-EF1α-EGFP vector backbone (Addgene, #60058, USA) at the KpnI and NcoI restriction sites using infusion cloning (Clontech, #639649, USA).

Techniques: Injection, Control