Journal: Mediators of Inflammation

Article Title: Continuous Theta-Burst Stimulation Promotes Paravascular CSF-Interstitial Fluid Exchange through Regulation of Aquaporin-4 Polarization in APP/PS1 Mice

doi: 10.1155/2022/2140524

Figure Lengend Snippet: CTBS improved AQP4 polarization in APP/PS1 mice. (a) Immunofluorescent staining of AQP4 in the cortex and hippocampus (200×, scale bar: 100 μ m). (b) Statistical analysis of relative AQP4 expression levels in the cortex. (c) Statistical analysis of relative AQP4 expression levels in the hippocampus. (d) Statistical analysis of relative perivascular polarization of AQP4 in the cortex. (e) Statistical analysis of relative perivascular polarization of AQP4 in the hippocampus. Data are presented as the mean ± SD ( n = 6 mice per group, unpaired t -test for comparing two individual groups; ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001, ∗∗∗∗ P < 0.0001).

Article Snippet: Antibodies for AQP4 (Alomone Labs, 300-314; Jerusalem, Israel) and glial fibrillary acidic protein (GFAP, Sigma, G3893, USA) were used to detect aquaporins and astrocytes.

Techniques: Staining, Expressing

Journal: Journal of Neuroscience Research

Article Title: Light microscopic and heterogeneity analysis of astrocytes in the common marmoset brain

doi: 10.1002/jnr.24967

Figure Lengend Snippet: List of primary antibodies used

Article Snippet: Anti‐AQP4 antibody produced in rabbit , Peptide corresponding to AA 300‐314 of rat AQP4 , Alomone lab, # AQP4‐014, polyclonal antibody, RRID:AB_11122614 , 1.6 μg/ml/1:500.

Techniques: Concentration Assay, Produced, Recombinant, Purification, Derivative Assay, Transduction

Journal: Journal of Neuroscience Research

Article Title: Light microscopic and heterogeneity analysis of astrocytes in the common marmoset brain

doi: 10.1002/jnr.24967

Figure Lengend Snippet: Antibodies used and amino acid conservation between species

Article Snippet: Anti‐AQP4 antibody produced in rabbit , Peptide corresponding to AA 300‐314 of rat AQP4 , Alomone lab, # AQP4‐014, polyclonal antibody, RRID:AB_11122614 , 1.6 μg/ml/1:500.

Techniques:

Journal: Journal of Neuroscience Research

Article Title: Light microscopic and heterogeneity analysis of astrocytes in the common marmoset brain

doi: 10.1002/jnr.24967

Figure Lengend Snippet: Heterogeneity of hippocampal astrocytes. Hierarchical clustering algorithm identified different astrocytic clusters based on the KS distance displayed in the dendrogram (left panel on each row) for GFAP (A), AQP4 (B), and Kir4.1 (C). The spatial organization of astrocytes within clusters is represented at the right side of each image (a–c). Scale bars: 50 μm. Each astrocyte was detected using GS expression and it was assigned in S. radiatum, S. pyramidale, and S. oriens. Each astrocyte was identified with a number, S. radiatum and S. oriens in white and S. pyramidale in yellow. Analysis of GS, EAAT2, and GAT3 can be found in Figure . Every astrocyte within a cluster was identified and assigned a number with a particular color. The percentage of cells in each cluster in the S. oriens (Ori), S. pyramidale (Pyr), and S. radiatum (Rad) is shown in a′–c′

Article Snippet: Anti‐AQP4 antibody produced in rabbit , Peptide corresponding to AA 300‐314 of rat AQP4 , Alomone lab, # AQP4‐014, polyclonal antibody, RRID:AB_11122614 , 1.6 μg/ml/1:500.

Techniques: Expressing

Journal: Journal of Neuroscience Research

Article Title: Light microscopic and heterogeneity analysis of astrocytes in the common marmoset brain

doi: 10.1002/jnr.24967

Figure Lengend Snippet: GFAP labels marmoset cortical astrocytic populations including complex interlaminar astrocytes. (A–E) Sparsely labeled GFAP‐positive astrocytes across cortical layers. The approximate location of cortical layers indicated by Roman numerals I to IV (yellow). Orange arrowheads show interlaminar processes and green arrowheads define protoplasmic astrocytes (A). Detailed images of an interlaminar astrocyte (B), protoplasmic astrocyte (C), astrocytic endfeet (D), and white matter astrocytes (E). DAPI (blue) is used as a nuclear marker and AQP4 (blue) as an endfoot marker. Scale bar: 50 μm (A), 20 μm (B, C, and E), and 5 μm (D). Subtypes of interlaminar astrocytes (ILA). Marmoset pial ILAs (F–J) and subpial ILAs (K–O) labeled by S100β (F, K), GS (G, L), Sox9 (H, M) and specific astrocytic markers such as Kir4.1 (I, N) and AQP4 (J, O). Yellow arrowheads show astrocytic soma and processes in F–O. Orange arrowheads describe a punctate AQP4 staining. Scale bars: 20 μm (F–O)

Article Snippet: Anti‐AQP4 antibody produced in rabbit , Peptide corresponding to AA 300‐314 of rat AQP4 , Alomone lab, # AQP4‐014, polyclonal antibody, RRID:AB_11122614 , 1.6 μg/ml/1:500.

Techniques: Labeling, Marker, Staining

Journal: Journal of Neuroscience Research

Article Title: Light microscopic and heterogeneity analysis of astrocytes in the common marmoset brain

doi: 10.1002/jnr.24967

Figure Lengend Snippet: Expression of glutamate and GABA transporters, Kir4.1, and AQP4 in the marmoset hippocampus. Labeling of coronal sections of marmoset hippocampus with astrocytic markers associated with subpopulations of astrocytes. Double immunolabeling of EAAT2 and GS (A), GAT3 and GS (B), Kir4.1 and GS (C) and water channel Aquaporin 4 (AQP4) and GS (D). The third panel of each row shows a merged image and the fourth panel is a heat map showing the density of expression of each specific marker. A purple–blue color indicates lower intensity signal and red–yellow color means higher intensity signal. White arrows highlight areas of particularly high signal intensity. Scale bars: 50 μm

Article Snippet: Anti‐AQP4 antibody produced in rabbit , Peptide corresponding to AA 300‐314 of rat AQP4 , Alomone lab, # AQP4‐014, polyclonal antibody, RRID:AB_11122614 , 1.6 μg/ml/1:500.

Techniques: Expressing, Labeling, Immunolabeling, Marker

Journal: Journal of Neuroscience Research

Article Title: Light microscopic and heterogeneity analysis of astrocytes in the common marmoset brain

doi: 10.1002/jnr.24967

Figure Lengend Snippet: Molecular heterogeneity of astrocytes in the marmoset cerebellum. (A) Lower magnification images showing labeling for astrocytic proteins with respect to the molecular layer (ML), Purkinje cell layer (PCL), granule cell layer (GCL), and white matter (WM). Calbindin is used as a Purkinje cell marker. DAPI staining (blue) shows the nuclear staining of interneurons in the ML and granule cells in GCL. Scale bar: 100 μm. (B) Detailed images of ML and GCL stained with specific markers for Bergmann glia (BG) such as GluA1, Kirk4.1, and velate astrocytes (VA) such as AQP4. EAAT2 was expressed in both BGs and VAs with higher expression in VAs. Purkinje cell bodies are indicated with a white asterisk. Scale bar: 20 μm. bv, blood vessel

Article Snippet: Anti‐AQP4 antibody produced in rabbit , Peptide corresponding to AA 300‐314 of rat AQP4 , Alomone lab, # AQP4‐014, polyclonal antibody, RRID:AB_11122614 , 1.6 μg/ml/1:500.

Techniques: Labeling, Marker, Staining, Expressing

Journal: Journal of Neuroscience Research

Article Title: Light microscopic and heterogeneity analysis of astrocytes in the common marmoset brain

doi: 10.1002/jnr.24967

Figure Lengend Snippet: Interactions of marmoset astrocytes with capillaries, neurons, and microglia. (A) Schema showing the gliovascular unit and blood vessel‐associated microglia. (B) GFAP and S100β labeling in astrocytic processes and endfeet surrounding a capillary in the marmoset cortex. Yellow circle shows high S100β expression and red circles highlight low S100β expression. Scale bar: 20 µm. (C) Cross‐sectional view of a capillary in the marmoset hippocampus displaying expression of AQP4 and GFAP at the astrocyte endfeet. Yellow arrowheads indicate GFAP expression at endfeet and white arrowheads indicate areas of co‐localization between GFAP and AQP4. Laminin (Lam) and DAPI labels show the basement membrane and nuclei of endothelial cells and pericytes (green arrowheads), respectively. Scale bars: 5 µm. (D) Cross‐sectional view of a capillary showing expression of Kir4.1 and GFAP at astrocyte endfeet and DAPI in endothelial cell and pericyte nuclei. Yellow arrowheads indicate GFAP expression, white arrowheads show areas of co‐localization between GFAP and Kir4.1, and green arrowheads highlight nuclei. Scale bars: 5 µm. (E–I) Co‐labeling of GFAP with neuronal markers including MAP2 (E, temporal cortex; G, hippocampus), Kv2.1 (F, temporal cortex; H hippocampus), and Calbindin (I, cerebellum). Higher magnification images are shown to the right of each panel (panels e′–i′). White arrowheads highlight the tight spatial neuronal‐astrocyte interaction. (J–L) Co‐labeling of GFAP with the microglial marker Iba1 in temporal cortex (J and K) and hippocampus (L). Images to the right of each panel (panels j′–l′) show higher magnifications. (J) Vessel‐associated microglia. Higher magnification images show microglia around the blood vessel (orange arrowheads) and microglia attached at the capillary (white arrowheads) (panels j′, j″). White arrowheads highlight the close spatial microglial–astrocyte interaction. Scale bars in panels E–L are 50 µm and e′–l′ are 20 µm. bv, blood vessel

Article Snippet: Anti‐AQP4 antibody produced in rabbit , Peptide corresponding to AA 300‐314 of rat AQP4 , Alomone lab, # AQP4‐014, polyclonal antibody, RRID:AB_11122614 , 1.6 μg/ml/1:500.

Techniques: Labeling, Expressing, Marker

Journal: Molecular Medicine Reports

Article Title: Alterations in AQP4 expression and polarization in the course of motor neuron degeneration in SOD1G93A mice

doi: 10.3892/mmr.2017.6786

Figure Lengend Snippet: GFAP and AQP4 expression and localization in the ventral horn of the lumbar spinal cord. (A) GFAP and AQP4 expression and localization in wild type control animals. (B) GFAP and AQP4 expression and localization in SOD1G93A mice at the presymptomatic stage. (C) GFAP and AQP4 expression and localization in SOD1G93A mice at the disease onset stage. (D) GFAP and AQP4 expression and localization in SOD1G93A mice at the end stage. The arrows indicate areas of positive AQP4 staining. Scale bars, 150 µm for the left three columns and 75 µm for the far right column. (E) GFAP and AQP4 expression was quantified at the presymptomatic, disease onset and end stages using the area fraction (percentage of GFAP or AQP4 immunoreactivity in the overall field). Increased global expression of GFAP and AQP4 was observed as the disease progressed. (F) AQP4 polarization decreased as the disease progressed. *P<0.05 vs. wild type control (one-way analysis of variance; n=5 animals/group). GFAP, glial fibrillary acidic protein; AQP4, aquaporin-4; SOD1, superoxide dismutase 1.

Article Snippet: Primary antibodies included goat anti-mouse choline O-acetyltransferase (ChAT) polyclonal antibody (cat. no. AB144P; EMD Millipore, Billerica, MA, USA) at 1:400 dilution; mouse anti-mouse glial fibrillary acidic protein (GFAP) monoclonal antibody (cat. no. 3670; Cell Signaling Technology, Inc., Danvers, MA, USA) at 1:500 dilution; rabbit anti-mouse AQP4 polyclonal antibody (cat. no. AQP-014; Alomone Labs, Jerusalem, Israel) at 1:400 dilution; and rabbit anti-mouse GLT-1 polyclonal antibody (cat. no. ab106289; Abcam, Cambridge, UK) at 1:300 dilution, overnight at 4°C.

Techniques: Expressing, Staining

Journal: Molecular Medicine Reports

Article Title: Alterations in AQP4 expression and polarization in the course of motor neuron degeneration in SOD1G93A mice

doi: 10.3892/mmr.2017.6786

Figure Lengend Snippet: GFAP and AQP4 expression and localization in the ventral horn of the cervical spinal cord. (A) GFAP and AQP4 expression and localization in control animals. (B) GFAP and AQP4 expression and localization in SOD1G93A mice at the presymptomatic stage. (C) GFAP and AQP4 expression and localization in SOD1G93A mice at the disease onset stage. (D) GFAP and AQP4 expression and localization in SOD1G93A mice at the end stage. The arrows indicate areas of positive AQP4 staining. Scale bars, 150 µm for the left three columns and 75 µm for the far right column. (E) GFAP and AQP4 expression was quantified at the presymptomatic, disease onset and end stages using the area fraction (percentage of GFAP or AQP4 immunoreactivity in the overall field). Increased global expression of GFAP and AQP4 was observed as the disease progressed. (F) AQP4 polarization decreased as the disease progressed. *P<0.05 vs. wild type control (one-way analysis of variance; n=5 animals/group). GFAP, glial fibrillary acidic protein; AQP4, aquaporin-4; SOD1, superoxide dismutase 1.

Article Snippet: Primary antibodies included goat anti-mouse choline O-acetyltransferase (ChAT) polyclonal antibody (cat. no. AB144P; EMD Millipore, Billerica, MA, USA) at 1:400 dilution; mouse anti-mouse glial fibrillary acidic protein (GFAP) monoclonal antibody (cat. no. 3670; Cell Signaling Technology, Inc., Danvers, MA, USA) at 1:500 dilution; rabbit anti-mouse AQP4 polyclonal antibody (cat. no. AQP-014; Alomone Labs, Jerusalem, Israel) at 1:400 dilution; and rabbit anti-mouse GLT-1 polyclonal antibody (cat. no. ab106289; Abcam, Cambridge, UK) at 1:300 dilution, overnight at 4°C.

Techniques: Expressing, Staining

Journal: Investigative Ophthalmology & Visual Science

Article Title: Rearing Light Intensity Affects Inner Retinal Pathology in a Mouse Model of X-Linked Retinoschisis but Does Not Alter Gene Therapy Outcome

doi: 10.1167/iovs.16-21016

Figure Lengend Snippet: Effect of light rearing on AQP-4, Kir4.1, and GFAP levels in 1- and 4-month-old Rs1 -KO and WT mice. Expression levels of AQP4, Kir4.1, and GFAP normalized to internal control protein actin are presented as mean ± standard error for 1- and 4-month-old Rs1 -KO and WT mice reared in either LL or ML. Rs1 -KO mice showed no significant difference in AQP4 and GFAP levels between LL and ML conditions at 1 and 4 months ( A , C ). Kir4.1 levels were unchanged between LL and ML Rs1 -KO mice at 1 month, but at 4 months they were significantly increased by 7-fold in ML-reared Rs1 -KO mice compared to those reared in LL ( B ). AQP4, Kir4.1, and GFAP levels were unchanged in WT mice irrespective of light conditions and age ( D – F ) but their levels were overall higher in Rs1 -KO mice than in WT animals, likely as a result of Müller cell response to the presence of intraretinal cavities. * P < 0.05. n.s., not significant.

Article Snippet: The membranes were incubated in Odyssey blocking buffer (LI-COR Biosciences, Lincoln, NE, USA) for 1 hour and later incubated overnight with one of the indicated primary antibodies diluted in PBS containing 0.05% Tween 20 (PBST), pH 7.5, at 4°C: anti–inward rectifier K+ ion channel 4.1 (Kir4.1, 1:400; Alomone Labs, Jerusalem, Israel), anti–water channel aquaporin-4 (AQP4, 1:1000; Alomone Labs), anti–glial fibrillary acidic protein (GFAP, 1:5000; Sigma-Aldrich Corp., St. Louis, MO, USA), and anti–β-actin (mouse monoclonal, 1:6000; Sigma-Aldrich).

Techniques: Expressing