Journal: The Journal of Neuroscience

Article Title: Elevated TRPV4 Levels Contribute to Endothelial Damage and Scarring in Experimental Spinal Cord Injury

doi: 10.1523/JNEUROSCI.2035-19.2020

Figure Lengend Snippet: Primary antibody information used for immunohistochemistry analysis

Article Snippet: The fluorescence IMV was plotted as mean ± SEM in GraphPad Prism (version 5.01, GraphPad Software). table ft1 table-wrap mode="anchored" t5 Table 3. caption a7 Primary antibody Dilution Producer company Code number Anti-TRPV4 1:500 Alomone Labs, Israel ACC-034 Anti-RECA-1 1:100 Abcam, UK ab22492 Anti-NF 1:1000 Abcam, UK ab8135 Anti-Iba-1 1:200 FUJIFILM Wako, Japan 019-19741 Anti-laminin 1:50 Sigma L9393 Anti-GFAP 1:500 Abcam, UK ab10062 Anti-ZO-1 1:50 Invitrogen 40-2300 Anti-occludin 1:50 Invitrogen 33-1500 Anti-collagen-IV 1:500 Abcam, UK ab6586 Anti-nitrotyrosine 1:100 Abcam, UK ab183391 Anti-CD-68 1:200 Abcam, UK ab955 Anti-TGF-β1 1:100 Abcam, UK ab25121 Anti-CD-206 1:500 Abcam, UK ab64693 Anti-BDNF 1:300 Alomone Labs, Israel AGP-021 Anti-NT-3 1:200 Alomone Labs, Israel ANT-003 Anti-vWF 1:50 Millipore AB7356 Anti-ANG-1 1:200 Abcam, UK ab133425 Anti-NG-2 1:500 Abcam, UK ab129051 Anti-α-SMA 1:200 Abcam, UK ab21027 Anti-connexin-43 1:500 Abcam, UK ab11369 Anti-Tuj-1 1:1000 Abcam, UK ab18207 Anti-growth cone 1:3 Abcam, UK ab7762 Anti-NeuN 1:1000 Abcam, UK ab177487 Open in a separate window Primary antibody information used for immunohistochemistry analysis table ft1 table-wrap mode="anchored" t5 Table 4. caption a7 Secondary antibody Dilution Producer company Code number Goat anti-rabbit AlexaFluor-488 1:200 Abcam, UK ab150077 Goat anti-rabbit AlexaFluor-568 1:200 Abcam, UK ab175471 Goat anti-rabbit AlexaFluor-647 1:200 Abcam, UK ab150083 Goat anti-mouse AlexaFluor-568 1:200 Abcam, UK ab175473 Goat anti-mouse AlexaFluor-488 1:200 Abcam, UK ab150117 Goat anti-guinea pig AlexaFluor-488 1:200 Abcam, UK ab150185 Chicken-anti-goat 647 1:200 Invitrogen, USA A-21469 Open in a separate window Secondary antibody information used for immunohistochemistry analysis Immunocytochemistry For immunofluorescence microscopy, third passage hCMEC/D3 BBB cell line and HUVECs (0.5 × 10 5 cells/well) were cultured on a sterile rat tail collagen-I (Sigma Millipore)-coated coverslip in 24-well plates.

Techniques: Immunohistochemistry

Journal: The Journal of Neuroscience

Article Title: Monocyte-Derived Macrophages Contribute to Spontaneous Long-Term Functional Recovery after Stroke in Mice

doi: 10.1523/JNEUROSCI.4317-15.2016

Figure Lengend Snippet: Primary antibodies used for immunocytochemistry

Article Snippet: Corresponding fluorophore-conjugated secondary antibodies (Invitrogen or The Jackson Laboratory) were diluted in blocking solution and applied for 2 h, followed by three rinses in potassium–PBS. table ft1 table-wrap mode="anchored" t5 Table 2. caption a7 Antibodies Host species Dilution Company BDNF Rabbit 1:200 Alomone Labs CD31 Rat 1:400 BD Pharmingen CD68 (ED1) Rat 1:200 AbD Serotec GFAP Rabbit 1:400 Zymed (Invitrogen) GFP Chicken 1:3000 Millipore IB4-FITC conjugated Lectin from Bandeiraea simplicifolia 1:50 Sigma-Aldrich Iba1 Goat 1:200 AbD Serotec IL-6 Rabbit 1:300 Abcam NeuN Mouse 1:100 Millipore TGFβ Mouse 1:300 R&D Systems Open in a separate window Primary antibodies used for immunocytochemistry Single labeling for NeuN was performed with biotinylated horse anti-mouse antibody and visualized with avidin–biotin–peroxidase complex (Elite ABC kit; Vector Laboratories), followed by peroxidase-catalyzed diaminobenzidine reaction.

Techniques:

Journal: The Journal of Neuroscience

Article Title: Monocyte-Derived Macrophages Contribute to Spontaneous Long-Term Functional Recovery after Stroke in Mice

doi: 10.1523/JNEUROSCI.4317-15.2016

Figure Lengend Snippet: Proinflammatory and anti-inflammatory factors are expressed in the stroke-injured hemisphere. A–C, Quantitative PCR shows increased expression in the injured (ipsilateral) hemisphere of anti-inflammatory factor (Ym1) at 3 d (A), proinflammatory (IL-6, TNFα, IL-1β, NOS2) and anti-inflammatory (TGFβ1, Ym1, CXCL113, CCL22, CD163) factors at 7 d (B), and anti-inflammatory factors (TGFβ1, VCAM1) at 14 d (C) after stroke (n = 7). Data are means ± SEMs; *p < 0.05, unpaired t test. D, Fluorescence microscopic images of CXCR3–GFP+ chimeric mouse brain coronal sections showing double-immunostaining of MDMs (green) and IL-6, TGFβ, and BDNF (all red) at 3 and 7 d after stroke. Note the decreased immunoreactivity for IL-6 and increased staining for TGFβ and BDNF at 7 d compared with 3 d. Scale bar, 150 μm.

Article Snippet: Corresponding fluorophore-conjugated secondary antibodies (Invitrogen or The Jackson Laboratory) were diluted in blocking solution and applied for 2 h, followed by three rinses in potassium–PBS. table ft1 table-wrap mode="anchored" t5 Table 2. caption a7 Antibodies Host species Dilution Company BDNF Rabbit 1:200 Alomone Labs CD31 Rat 1:400 BD Pharmingen CD68 (ED1) Rat 1:200 AbD Serotec GFAP Rabbit 1:400 Zymed (Invitrogen) GFP Chicken 1:3000 Millipore IB4-FITC conjugated Lectin from Bandeiraea simplicifolia 1:50 Sigma-Aldrich Iba1 Goat 1:200 AbD Serotec IL-6 Rabbit 1:300 Abcam NeuN Mouse 1:100 Millipore TGFβ Mouse 1:300 R&D Systems Open in a separate window Primary antibodies used for immunocytochemistry Single labeling for NeuN was performed with biotinylated horse anti-mouse antibody and visualized with avidin–biotin–peroxidase complex (Elite ABC kit; Vector Laboratories), followed by peroxidase-catalyzed diaminobenzidine reaction.

Techniques: Real-time Polymerase Chain Reaction, Expressing, Fluorescence, Double Immunostaining, Staining

Journal:

Article Title: Brain-derived neurotrophic factor (BDNF) contributes to neuronal dysfunction in a model of allergic airway inflammation

doi: 10.1038/sj.bjp.0705638

Figure Lengend Snippet: Animal treatment protocol. Mice were sensitised to OVA adsorbed to Al(OH)3 or vehicle alone by intraperitoneal injections on days 1, 14 and 21. Prior to analysis, animals received two consecutive local aerosol challenges of 1% OVA (w v−1) diluted in PBS or PBS alone and delivered by 20-min aerosolisation on days 26 and 27. Intranasal application of polyclonal chicken IgY (isotype antibody) or anti-mouse BDNF was performed 3 h before each airway allergen challenge. In addition, animals received the antibodies i.p. on day 25. The response to acute allergen exposure was measured on day 35 in the body plethysmograph. All animals were analysed 24 h after the last challenge. Abbreviations: intraperitoneally (i.p.), intranasally (i.n.), aerosol challenge (aerosol).

Article Snippet: Polyclonal chicken IgY anti-mouse BDNF (2 × 25 μ l) (500 μ g ml −1 ) or isotype antibody (500 μ g ml −1 ) (Promega, Madison, U.S.A.) solved in sterile PBS was instilled intranasally 3 h before each airway allergen challenge.

Techniques: Aerosol

Journal:

Article Title: Brain-derived neurotrophic factor (BDNF) contributes to neuronal dysfunction in a model of allergic airway inflammation

doi: 10.1038/sj.bjp.0705638

Figure Lengend Snippet: Overview of experiments and animal numbers

Article Snippet: Polyclonal chicken IgY anti-mouse BDNF (2 × 25 μ l) (500 μ g ml −1 ) or isotype antibody (500 μ g ml −1 ) (Promega, Madison, U.S.A.) solved in sterile PBS was instilled intranasally 3 h before each airway allergen challenge.

Techniques: Aerosol

Journal:

Article Title: Brain-derived neurotrophic factor (BDNF) contributes to neuronal dysfunction in a model of allergic airway inflammation

doi: 10.1038/sj.bjp.0705638

Figure Lengend Snippet: Effect of anti-BDNF treatment on allergen-induced airway obstruction by HBP. Measurement of expiratory flow in response to OVA (intranasal application during continuous measurement of airflow in the HBP). (a) Integration of EF50 values between 15 and 25 min after OVA application. (b) Time course of n=11 animals in each group. Data from three separate experiments are shown. Grey bar: possible artefacts from OVA application. The error bars represent standard error of the mean (s.e.m.). Student́s t-test: ***P<0.001.

Article Snippet: Polyclonal chicken IgY anti-mouse BDNF (2 × 25 μ l) (500 μ g ml −1 ) or isotype antibody (500 μ g ml −1 ) (Promega, Madison, U.S.A.) solved in sterile PBS was instilled intranasally 3 h before each airway allergen challenge.

Techniques:

Journal:

Article Title: Brain-derived neurotrophic factor (BDNF) contributes to neuronal dysfunction in a model of allergic airway inflammation

doi: 10.1038/sj.bjp.0705638

Figure Lengend Snippet: Effect of BDNF and anti-BDNF treatment on inflammation. (a, b) Cytokine levels in BALF. IL-4, IL-5 and IFN-γ were measured in BALF from nonsensitised or OVA-sensitised mice 24 h after aerosol challenge by ELISA. The BAL recovery was 1.4±0.2 ml in all groups. (c, d) Cell differentiation in BALF after OVA aerosol challenge on day 28. Lymphocytes, macrophages, eosinophils and neutrophils were differentiated according to morphological criteria in BALF from nonsensitised or OVA-sensitised mice 24 h after the last aerosol challenge. (e) Cell differentiation in BALF 24 h after nasal OVA challenge on day 36. Student's t-test did not reveal any statistically significant difference (P<0.05) between groups. In all, 12 mice were analysed in each group, in two separate experiments. The error bars represent standard deviation (s.d.).

Article Snippet: Polyclonal chicken IgY anti-mouse BDNF (2 × 25 μ l) (500 μ g ml −1 ) or isotype antibody (500 μ g ml −1 ) (Promega, Madison, U.S.A.) solved in sterile PBS was instilled intranasally 3 h before each airway allergen challenge.

Techniques: Aerosol, Enzyme-linked Immunosorbent Assay, Cell Differentiation, Standard Deviation

Journal:

Article Title: Brain-derived neurotrophic factor (BDNF) contributes to neuronal dysfunction in a model of allergic airway inflammation

doi: 10.1038/sj.bjp.0705638

Figure Lengend Snippet: Effect of BDNF on airway hyper-responsiveness in response to EFS. Airway hyper-responsiveness was measured in response to EFS. The frequency that caused 50% of maximal airway smooth muscle constriction was defined as ES50. Airway contractility was expressed as mean±s.d. Statistics were performed using t-test with Welch's correction. P<0.05 was regarded as a statistically significant difference. (a) Nonsensitised BALB/c mice were treated intranasally either with recombinant human BDNF (5 μg ml−1, 50 μl) or BSA (0.1%, 50 μl) as a control on two consecutive days. AHR was measured 24 h after the last treatment. In all, 12 animals were examined in each group in two separate experiments. (a) OVA-sensitised Balb/c mice were treated intranasally either with anti-BDNF (500 μg ml−1, 50 μl) or istotype IgY (500 μg ml−1, 50 μl) as a control on two consecutive days, 3 h before OVA aerosol challenge. In addition, the animals received 100 μl of the antibody solution i.p. 24 h before the first OVA challenge (day 25). AHR was measured 24 h after the last treatment. A total of 12 animals were examined in each group in two separate experiments.

Article Snippet: Polyclonal chicken IgY anti-mouse BDNF (2 × 25 μ l) (500 μ g ml −1 ) or isotype antibody (500 μ g ml −1 ) (Promega, Madison, U.S.A.) solved in sterile PBS was instilled intranasally 3 h before each airway allergen challenge.

Techniques: Recombinant, Control, Aerosol

Journal:

Article Title: Brain-derived neurotrophic factor (BDNF) contributes to neuronal dysfunction in a model of allergic airway inflammation

doi: 10.1038/sj.bjp.0705638

Figure Lengend Snippet: Effect of anti-BDNF treatment on airway hyper-responsiveness in response to methacholine and capsaicin by HBP. (a) Dose–response curve of expiratory flow (EF50) in response to inhaled methacholine. (b) Dose–response curve of expiratory flow (EF50) in response to inhaled capsaicin. (c) Dose–response curve of sensory irritation (TB) in response to inhaled methacholine. (d) Dose–response curve of sensory irritation (TB) in response to inhaled capsaicin. The error bars represent standard error of the mean (s.e.m.), n=8–10 animals in each group in three separate experiments. Student's t-test: *P<0.05. (a) Nonsensitised against OVA-sensitised+isotype, (d) OVA-sensitised+isotype against OVA-sensitised+anti-BDNF.

Article Snippet: Polyclonal chicken IgY anti-mouse BDNF (2 × 25 μ l) (500 μ g ml −1 ) or isotype antibody (500 μ g ml −1 ) (Promega, Madison, U.S.A.) solved in sterile PBS was instilled intranasally 3 h before each airway allergen challenge.

Techniques:

Journal:

Article Title: Brain-derived neurotrophic factor (BDNF) contributes to neuronal dysfunction in a model of allergic airway inflammation

doi: 10.1038/sj.bjp.0705638

Figure Lengend Snippet: Tachykinin containing sensory neurons from ovalbumin-sensitised mice

Article Snippet: Polyclonal chicken IgY anti-mouse BDNF (2 × 25 μ l) (500 μ g ml −1 ) or isotype antibody (500 μ g ml −1 ) (Promega, Madison, U.S.A.) solved in sterile PBS was instilled intranasally 3 h before each airway allergen challenge.

Techniques:

Journal: Frontiers in Molecular Neuroscience

Article Title: Evaluation of Biochemical and Epigenetic Measures of Peripheral Brain-Derived Neurotrophic Factor (BDNF) as a Biomarker in Huntington’s Disease Patients

doi: 10.3389/fnmol.2019.00335

Figure Lengend Snippet: Plasma BDNF levels according to sex and diagnostic group. BDNF levels in plasma (50 ul) were determined by ELISA. (A) BDNF levels did not differ by sex among subjects included in our study as per Student’s t test, unpaired; two-tailed. (B) No significant differences were detected according to diagnostic group between premanifest (PM), manifest HD (HD), and normal controls (NC) as per One-way ANOVA. Data is shown as individual BDNF levels ± SD.

Article Snippet: Subsequently, a polyclonal chicken anti-human BDNF (100 ul; 2.5 μg/ml; Promega) was added to the plate and incubated for 2.5 h. The plate was washed five times with PBST, then anti-chicken IgY-HRP (1 μg/ml; Promega) was added to each well for a 1-h incubation at RT.

Techniques: Clinical Proteomics, Diagnostic Assay, Enzyme-linked Immunosorbent Assay, Two Tailed Test

Journal: Frontiers in Molecular Neuroscience

Article Title: Evaluation of Biochemical and Epigenetic Measures of Peripheral Brain-Derived Neurotrophic Factor (BDNF) as a Biomarker in Huntington’s Disease Patients

doi: 10.3389/fnmol.2019.00335

Figure Lengend Snippet: Correlations between plasma BDNF levels and clinical data in HD patients and control subjects.

Article Snippet: Subsequently, a polyclonal chicken anti-human BDNF (100 ul; 2.5 μg/ml; Promega) was added to the plate and incubated for 2.5 h. The plate was washed five times with PBST, then anti-chicken IgY-HRP (1 μg/ml; Promega) was added to each well for a 1-h incubation at RT.

Techniques: Clinical Proteomics, Control

Journal: Frontiers in Molecular Neuroscience

Article Title: Evaluation of Biochemical and Epigenetic Measures of Peripheral Brain-Derived Neurotrophic Factor (BDNF) as a Biomarker in Huntington’s Disease Patients

doi: 10.3389/fnmol.2019.00335

Figure Lengend Snippet: Salivary BDNF levels according to sex and diagnostic group. BDNF levels in saliva (50 ul) were determined by ELISA. (A) No significant differences in BDNF were observed by sex (Student’s t test, unpaired; two-tailed). (B) BDNF levels are lower in premanifest (PM) and manifest HD (HD) cases when compared to normal controls (NC). ∗ p < 0.05; ∗∗ p < 0.01 in comparison to NC group as per One-way ANOVA. (C) BDNF levels are significantly lower in subjects who are >10 years from their predicted age of onset compared to those who are <10 years from predicted onset. ∗ p = 0.05. Predicted age to onset was calculated using the Langbehn formula . Data is shown as individual BDNF levels ± SD.

Article Snippet: Subsequently, a polyclonal chicken anti-human BDNF (100 ul; 2.5 μg/ml; Promega) was added to the plate and incubated for 2.5 h. The plate was washed five times with PBST, then anti-chicken IgY-HRP (1 μg/ml; Promega) was added to each well for a 1-h incubation at RT.

Techniques: Diagnostic Assay, Enzyme-linked Immunosorbent Assay, Two Tailed Test, Comparison

Journal: Frontiers in Molecular Neuroscience

Article Title: Evaluation of Biochemical and Epigenetic Measures of Peripheral Brain-Derived Neurotrophic Factor (BDNF) as a Biomarker in Huntington’s Disease Patients

doi: 10.3389/fnmol.2019.00335

Figure Lengend Snippet: Correlations between salivary BDNF levels and clinical data.

Article Snippet: Subsequently, a polyclonal chicken anti-human BDNF (100 ul; 2.5 μg/ml; Promega) was added to the plate and incubated for 2.5 h. The plate was washed five times with PBST, then anti-chicken IgY-HRP (1 μg/ml; Promega) was added to each well for a 1-h incubation at RT.

Techniques:

Journal: Frontiers in Molecular Neuroscience

Article Title: Evaluation of Biochemical and Epigenetic Measures of Peripheral Brain-Derived Neurotrophic Factor (BDNF) as a Biomarker in Huntington’s Disease Patients

doi: 10.3389/fnmol.2019.00335

Figure Lengend Snippet: Methylation sites in the BDNF promoter IV region. (A) Schematic representation of human BDNF gene indicating alternative promoters. CpG sites analyzed are detailed in enlarged box. Position is in relation to transcription start site. (B) Detailed sequence of BDNF promoter IV. Underlined text in yellow boxes indicate regions analyzed with each probe by pyrosequencing. Genomic position is based on chromone build GRCh37.p13 primary assembly.

Article Snippet: Subsequently, a polyclonal chicken anti-human BDNF (100 ul; 2.5 μg/ml; Promega) was added to the plate and incubated for 2.5 h. The plate was washed five times with PBST, then anti-chicken IgY-HRP (1 μg/ml; Promega) was added to each well for a 1-h incubation at RT.

Techniques: Methylation, Sequencing

Journal: Frontiers in Molecular Neuroscience

Article Title: Evaluation of Biochemical and Epigenetic Measures of Peripheral Brain-Derived Neurotrophic Factor (BDNF) as a Biomarker in Huntington’s Disease Patients

doi: 10.3389/fnmol.2019.00335

Figure Lengend Snippet: DNA methylation status at selected CpG sites from BDNF promoter IV.

Article Snippet: Subsequently, a polyclonal chicken anti-human BDNF (100 ul; 2.5 μg/ml; Promega) was added to the plate and incubated for 2.5 h. The plate was washed five times with PBST, then anti-chicken IgY-HRP (1 μg/ml; Promega) was added to each well for a 1-h incubation at RT.

Techniques: DNA Methylation Assay

Journal: Frontiers in Molecular Neuroscience

Article Title: Evaluation of Biochemical and Epigenetic Measures of Peripheral Brain-Derived Neurotrophic Factor (BDNF) as a Biomarker in Huntington’s Disease Patients

doi: 10.3389/fnmol.2019.00335

Figure Lengend Snippet: DNA methylation levels of BDNF promoter IV in blood. Methylation was quantified by pyrosequencing and expressed as mean percent methylation representing the averaged value of cases (HD Gene+) or control subjects (NC) for each of individual CpG. Data represents mean value ± SEM. ∗ p < 0.05 as per two-tailed Student’s t -test.

Article Snippet: Subsequently, a polyclonal chicken anti-human BDNF (100 ul; 2.5 μg/ml; Promega) was added to the plate and incubated for 2.5 h. The plate was washed five times with PBST, then anti-chicken IgY-HRP (1 μg/ml; Promega) was added to each well for a 1-h incubation at RT.

Techniques: DNA Methylation Assay, Methylation, Control, Two Tailed Test

Journal: Frontiers in Molecular Neuroscience

Article Title: Evaluation of Biochemical and Epigenetic Measures of Peripheral Brain-Derived Neurotrophic Factor (BDNF) as a Biomarker in Huntington’s Disease Patients

doi: 10.3389/fnmol.2019.00335

Figure Lengend Snippet: Correlations between BDNF promoter IV methylation and clinical measures in HD patients.

Article Snippet: Subsequently, a polyclonal chicken anti-human BDNF (100 ul; 2.5 μg/ml; Promega) was added to the plate and incubated for 2.5 h. The plate was washed five times with PBST, then anti-chicken IgY-HRP (1 μg/ml; Promega) was added to each well for a 1-h incubation at RT.

Techniques: Methylation