Journal: Journal of Advanced Research

Article Title: Aquaporin 4 and its isoforms regulation ameliorate AQP4 Mis-localization-induced glymphatic dysfunction in ischemic stroke

doi: 10.1016/j.jare.2025.05.022

Figure Lengend Snippet: Dynamic Evaluation of tMCAO Brain Injury and Glymphatic System Function in Mice Over Time. A Overview of the experimental process and group allocations. B Representative T2WI and DWI scans of the sham group and ischemic stroke group at various time points. C Ischemic lesion and brain swelling volumes in sham and tMCAO group at indicated time points (n = 8 mice per group). D Representative Western blots of AQP4 protein expression. E Quantification of relative AQP4 protein expression normalized (n = 8 mice per group). F Representative sagittal MRI images showing the flow of the contrast agent Gd-DTPA. G-I Quantitative analysis of signal-to-noise ratio variations and AUC in three ROIs across a 210-minute DCE Sequence (n = 4 mice per group). All data are shown as mean ± SD. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001 vs. sham;^P < 0.05,^^P < 0.05,^^^P < 0.001,^^^^P < 0.0001 vs. tMCAO 2 h; #P < 0.05, ##P < 0.01 vs. tMCAO 1 d; &&&P < 0.001 vs. tMCAO 3 d). All data were compared by one-way ANOVA with Tukey’s post hoc test.

Article Snippet: The AQP4 inhibitor TGN-020 (0.1 mg/20 g, HY-W008574, MedChemExpress,) was dissolved in 0.5 % CMC-Na (HY-Y1889A, MedChemExpress) and administered via intraperitoneal injection every 12 h, starting 10 min post-occlusion, as previously described [ ].

Techniques: Western Blot, Expressing, Sequencing

Journal: Journal of Advanced Research

Article Title: Aquaporin 4 and its isoforms regulation ameliorate AQP4 Mis-localization-induced glymphatic dysfunction in ischemic stroke

doi: 10.1016/j.jare.2025.05.022

Figure Lengend Snippet: TGN-020 Improves Brain Injury and Glymphatic System Dysfunction in tMCAO Mice. A Overview of the second part of the experimental process and group allocations. B Representative T2WI and DWI images of three distinct mouse groups. C Ischemic lesion volumes and brain swelling volumes in each group (n = 5 mice per group). D Schematic of intracerebral injection procedure in mice. Representative fluorescence microscopy images show Ovalbumin (45-kDa, red) inflow into CSF at 30- and 60-minute. The white arrow indicates the residual tracer deposition. Scale bar: 1000 μm. E Quantification of Ovalbumin inflow at 30- and 60-minute in brain sections (n = 4 mice per group). F Brain slices from each group show dextran (3-kDa, green) and ovalbumin (45-kDa, yellow) inflow into the brain cistern. Scale bar: 1000 μm. G Quantification of Glucan and Ovalbumin inflow in brain sections from each group (n = 4 mice per group). H Visual depiction of interstitial drainage (highlighted in red) following the introduction of the Ovalbumin tracer into the infarcted side. Scale bar: 1000 μm. I Quantification of the residual Ovalbumin tracer-covered area fraction in each group (n = 5 mice per group). All data are shown as mean ± SD. *P < 0.05, ****P < 0.0001 vs. sham; #P < 0.05, ###P < 0.001, ####P < 0.0001 vs. tMCAO + vehicle. One-way ANOVA with Tukey’s multiple comparisons test was performed in (C) and (I). Two-way repeated-measures ANOVA with Tukey’s post hoc test in (E) and (G).

Article Snippet: The AQP4 inhibitor TGN-020 (0.1 mg/20 g, HY-W008574, MedChemExpress,) was dissolved in 0.5 % CMC-Na (HY-Y1889A, MedChemExpress) and administered via intraperitoneal injection every 12 h, starting 10 min post-occlusion, as previously described [ ].

Techniques: Injection, Fluorescence, Microscopy

Journal: Journal of Advanced Research

Article Title: Aquaporin 4 and its isoforms regulation ameliorate AQP4 Mis-localization-induced glymphatic dysfunction in ischemic stroke

doi: 10.1016/j.jare.2025.05.022

Figure Lengend Snippet: TGN-020 Improves the Structure of Intracerebral Transport in the Glymphatic and Meningeal Lymphatic Systems. A Comparison of GFAP-positive areas (green) and AQP4 polarization (red) in the peri -infarct cortex. Scale bar, 50 μm. B Quantification of AQP4 polarization across the three groups (n = 6 mice per group). C-E Representative Western blot bands and densitometric quantification of AQP4-M1 and AQP4-M23 in the peri -infarction area (n = 6 per group). F Perivascular space structure observed by HE staining. TEM images showing perivascular astrocyte end-feet (green) in each group. H Representative images of meningeal lymphatic vessels stained with LYVE1. The enlarged view shows meningeal lymphatic vessels in the cortical subarachnoid space (COS) region. H-I Quantification of LYVE1 distribution and vessel diameter in the three groups (n = 6 mice per group). All data are shown as mean ± SD. *P < 0.05, ****P < 0.0001 vs. sham; ##P < 0.01 vs. tMCAO + vehicle. ALL data were compared by one-way ANOVA with Tukey’s post hoc test. AC, astrocytes; drAC, detached retracted astrocytes; EC, endothelial cell; RBC, red blood cell.

Article Snippet: The AQP4 inhibitor TGN-020 (0.1 mg/20 g, HY-W008574, MedChemExpress,) was dissolved in 0.5 % CMC-Na (HY-Y1889A, MedChemExpress) and administered via intraperitoneal injection every 12 h, starting 10 min post-occlusion, as previously described [ ].

Techniques: Comparison, Western Blot, Staining

Journal: Journal of Advanced Research

Article Title: Aquaporin 4 and its isoforms regulation ameliorate AQP4 Mis-localization-induced glymphatic dysfunction in ischemic stroke

doi: 10.1016/j.jare.2025.05.022

Figure Lengend Snippet: Effects of TGN-020 on cognitive dysfunction and metabolic pattern in tMCAO mice. A Representative image of swim traces from the Morris water maze test. B-D Data from the MWM test analyzing spatial learning and memory abilities of tMCAO mice (n = 10 mice per group). E Representative thermal imaging from the probe trial in the novel object recognition test. F Relative cognitive index with the novel object analyzed to evaluate memory ability following tMCAO (n = 10 mice per group). G Duration and speed in the rotarod test used to assess motor function in each group (n = 10 mice per group). H Transcriptomic changes in tMCAO mice. Red indicates down-regulated genes, blue dots indicate up-regulated genes, and gray dots indicate no significant changes. There was no significant change in the expression of AQP4 and SNTA1. I GSEA shows the ubiquitin-activity pathway. J Workflow for metabolomics analysis. K Volcano plot comparing differential metabolites between the tMCAO + vehicle and tMCAO + TGN-020 groups. L Pie chart depicting the proportions of different metabolite categories between the tMCAO + vehicle and tMCAO + TGN-020 groups. M Heatmap of the top 20 differentially expressed metabolites. N Analysis of affected metabolic pathways in comparisons between the tMCAO + vehicle and tMCAO + TGN-020 groups. All data are shown as mean ± SD. ****P < 0.0001 vs. sham; #P < 0.05, ##P < 0.01, ####P < 0.0001 vs. tMCAO + vehicle. One-way ANOVA with Tukey’s multiple comparisons test was performed in (B), (C) and (F). Two-way repeated-measures ANOVA with Tukey’s post hoc test in (D).

Article Snippet: The AQP4 inhibitor TGN-020 (0.1 mg/20 g, HY-W008574, MedChemExpress,) was dissolved in 0.5 % CMC-Na (HY-Y1889A, MedChemExpress) and administered via intraperitoneal injection every 12 h, starting 10 min post-occlusion, as previously described [ ].

Techniques: Imaging, Expressing, Ubiquitin Proteomics, Activity Assay

Journal: Journal of Advanced Research

Article Title: Aquaporin 4 and its isoforms regulation ameliorate AQP4 Mis-localization-induced glymphatic dysfunction in ischemic stroke

doi: 10.1016/j.jare.2025.05.022

Figure Lengend Snippet: Different AQP4 Isoforms Affect Brain Injury and Glymphatic Function in tMCAO Mice. A Overview of the third part of the experimental process and group allocations. B Relative mRNA expression levels of two isoforms of AQP4 in the peri -infarction area after tMCAO (n = 6 mice per group). C Representative T2 and DWI images of the three distinct groups of mice. D Ischemic lesion volumes and brain swelling volumes in each group (n = 6 mice per group). E Description of brain injection techniques and representative brain slice staining from three groups showing dextran (3-kDa, green) inflow into the brain cistern (above) and parenchyma (down) at 30 min. Scale bar: 1000 μm. F Quantification of Glucan inflow into the cistern (n = 3 mice per group). G Quantification of the residual Glucan in each group (n = 3 mice per group). H Comparison of GFAP-positive areas (green), AQP4 (red), and CD31 (grey) in the peri -infarct cortex. Scale bar: 100 μm. I Quantification of AQP4 polarization across the three groups (n = 4 mice per group). J Representative thermal imaging from the probe trial in the NOR test. K Relative cognitive index with the novel object analyzed to evaluate memory ability (n = 10 mice per group). L Duration and speed in the rotarod test used to assess motor function in each group (n = 10 mice per group). All data are shown as mean ± SD. *P < 0.05, **P < 0.01, ****P < 0.01 vs. tMCAO + AAV-CTRL; #P < 0.05, ##P < 0.01, ####P < 0.0001 vs. tMCAO + AAV-AQP4-M1. All data were compared by one-way ANOVA with Tukey’s post hoc test.

Article Snippet: The AQP4 inhibitor TGN-020 (0.1 mg/20 g, HY-W008574, MedChemExpress,) was dissolved in 0.5 % CMC-Na (HY-Y1889A, MedChemExpress) and administered via intraperitoneal injection every 12 h, starting 10 min post-occlusion, as previously described [ ].

Techniques: Expressing, Injection, Slice Preparation, Staining, Comparison, Imaging

Journal: Journal of Advanced Research

Article Title: Aquaporin 4 and its isoforms regulation ameliorate AQP4 Mis-localization-induced glymphatic dysfunction in ischemic stroke

doi: 10.1016/j.jare.2025.05.022

Figure Lengend Snippet: SNTA1 overexpression improves glymphatic clearance in tMCAO mice. A Overview of the last part of the experimental process and group allocations. B-F Representative Western blot bands and densitometric quantification of AQP4-M23, AQP4-M1, SNTA1 and AQP4-M23/AQP4-M1 in the peri -infarction area (n = 8 per group). G Co-IP of SNTA1 with AQP4 from tMCAO mice brain precipitated by the SNTA1 antibody. H Description of brain injection techniques and representative brain slice staining showing ovalbumin (45-kDa, green) inflow into the brain cistern (left) and parenchyma (right) at 30 min. Scale bar:1000 μm. I Quantification of Ovalbumin inflow into the cistern (n = 3 mice per group). J Quantification of residual Ovalbumin-covered area fraction (n = 3 mice per group). K Comparison of SNTA1(green), AQP4 polarization (red), and GFAP (grey) in the peri -infarct cortex. Scale bar: 200 μm. L Quantification of AQP4 polarization across the four groups (n = 3 mice per group). All data are shown as mean ± SD. **P < 0.01, vs. tMCAO + shCTRL; #P < 0.05, ##P < 0.01 vs tMCAO + AAV-CTRL. All data were compared by one-way ANOVA with Tukey’s post hoc test.

Article Snippet: The AQP4 inhibitor TGN-020 (0.1 mg/20 g, HY-W008574, MedChemExpress,) was dissolved in 0.5 % CMC-Na (HY-Y1889A, MedChemExpress) and administered via intraperitoneal injection every 12 h, starting 10 min post-occlusion, as previously described [ ].

Techniques: Over Expression, Western Blot, Co-Immunoprecipitation Assay, Injection, Slice Preparation, Staining, Comparison

Journal: Alzheimer's & Dementia

Article Title: Plasma extracellular vesicles from APOE3 Christchurch carriers display a protective phenotype in early stages of autosomal dominant Alzheimer's disease

doi: 10.1002/alz.71148

Figure Lengend Snippet: APOE3 Ch is associated with reduced pericyte and astrocyte EV markers alongside increased CD171+ EVs. A, Percentage of EVs positive for PDGFRβ (pericytes), AQP4 (astrocytes), CD171, and double‐positive PDGFRβ‐AQP4. B, Normalized mean fluorescence intensity (MFI) of EVs positive for markers in (A). C, Heatmap showing EVs positivity for the marker panel in (A), comparing PSEN1 E280A ‐APOE3 carriers with and without MCI. D, Heatmap of MFI for PSEN1 E280A ‐APOE3 carriers with and without MCI. E, Violin plots comparing positivity of markers in non‐MCI PSEN1 E280A carriers to APOE3 and APOE3 Ch . F, Violin plots comparing the MFI in non‐MCI PSEN1 E280A carriers to APOE3 and APOE3Ch . In (C) and (D), each column in the heatmap is labeled with a number, where each number represents an individual patient. Representative data from CNT, n = 10 (C1, C2, C3, C4, C5, C6, C7, C8, C9, C10); APOE3 Ch , n = 10 (Ch1, Ch2, Ch3, Ch4, Ch5, Ch6, Ch7, Ch8, Ch9, Ch10); MCI PSEN1 E280A ‐APOE3 , n = 9 (MCI1, MCI2, MCI3, MCI4, MCI5, MCI6, MCI7, MCI9, MCI10); non‐MCI PSEN1 E280A ‐APOE3 , n = 8 (non‐MCI‐1, non‐MCI‐2, non‐MCI‐3, non‐MCI‐4, non‐MCI‐5, non‐MCI‐7, non‐MCI‐8, non‐MCI‐9) and non‐MCI PSEN1 E280A ‐APOE3 Ch , n = 9 (E‐Ch1, E‐Ch2, E‐Ch3, E‐Ch4, E‐Ch5, E‐Ch6, E‐Ch7, E‐Ch8, E‐Ch9). Data are presented as boxplots or violin plots. Boxplots display the median, interquartile range, and whiskers representing minimum and maximum values. Violin plots illustrate the full data distribution, with individual data points overlaid and a line indicating the median. The data for the homozygous APOE3 Ch patient is indicated by a red dot. One‐way analysis of variance, Tukey multiple comparison test; unpaired t test, Mann–Whitney test were performed. Comparison to CNT is denoted by number sign (#); with non‐MCI PSEN1 E280A ‐APOE3 by asterisk (*). */# indicates P < 0.05, **/## P < 0.01, and ***/### P < 0.001. APOE , apolipoprotein E; AQP4, aquaporin 4; CD, cluster of differentiation; CNT, cognitively unimpaired control; EV, extracellular vesicle; MCI, mild cognitive impairment; MFI, mean fluorescence intensity; PDGFRβ, platelet‐derived growth factor beta; PSEN, presenilin

Article Snippet: The following markers were used: anti‐CD45 FITC (Clone HI30, BD Biosciences, Table ), anti‐CD105 PE (Clone 43A3, BioLegend, Table ), anti‐CD235a Pacific Blue (Clone HI264, BioLegend, Table ), anti‐CD41a FITC (Clone HIP8, BioLegend, Table ), anti‐aquaporin 4 (AQP4) FITC (Bioss, Table ), anti‐CD171 BV421 (Clone 5G3, BD, Table ), and anti‐CD140b (PDGFRβ) PE‐Cy7 (Clone APB5, Invitrogen, Table ).

Techniques: Fluorescence, Marker, Labeling, Comparison, MANN-WHITNEY, Control, Derivative Assay