rat l type vdcc  (Alomone Labs)

Journal: The Journal of Neuroscience

Article Title: Visual Experience Facilitates BDNF-Dependent Adaptive Recruitment of New Neurons in the Postembryonic Optic Tectum

doi: 10.1523/JNEUROSCI.1962-17.2018

Figure Lengend Snippet: Rearing larvae in dim light reduces production of BDNF in the medial PGZ by 10 dpf ( A–C ). Scale bar, 15 μm. Injecting 5 dpf larvae with 100 ng/ml recombinant human BDNF increased the amount of BDNF protein detected by immunohistochemistry

Article Snippet: To test whether exogenous BDNF was detectable in whole-brain lysates following ICV injections, we performed a Western blot using 5 dpf larvae injected with either 100 ng/ml recombinant human BDNF protein (#B-250, Alomone Labs) or a vehicle control.

Techniques: Recombinant, Immunohistochemistry

Journal: Journal of Neuroinflammation

Article Title: ProBDNF promotes sepsis-associated encephalopathy in mice by dampening the immune activity of meningeal CD4+ T cells

doi: 10.1186/s12974-020-01850-0

Figure Lengend Snippet: Schematic diagram showing how proBDNF dampens CD4 + T cell activity and contributes to the pathogenesis of SAE. In sepsis, proBDNF expression is increased in peripheral blood and meningeal immune cells, which then decreases the infiltration of CD4 + T cells in the meninges. As a result, meningeal pro-inflammatory cytokines such as IL-6 and IL-1β are upregulated, but anti-inflammatory cytokines including IL-4 and IL-13 are downregulated, finally leading to SAE. SAE, sepsis-associated encephalopathy

Article Snippet: 4 × 105 cells were put in each well of a 96-well flat-bottom plate and stimulated with 100, 200, or 500 ng ml−1 proBDNF protein (Alomone Labs, Israel, catalog: B243) as introduced by our previous studies [ ], respectively.

Techniques: Activity Assay, Expressing

Journal: Journal of Neuroinflammation

Article Title: ProBDNF promotes sepsis-associated encephalopathy in mice by dampening the immune activity of meningeal CD4+ T cells

doi: 10.1186/s12974-020-01850-0

Figure Lengend Snippet: Exogenous proBDNF protein reduced CD4 + T cells but increased CD8 + T cells of septic mice in vitro. Mice injected with saline or LPS (5 mg kg −1 ) for 5 days and the splenocytes were isolated and cultured for 3 days in vitro. Exogenous proBDNF did not alter the percentage of a CD3 + T cells in CD45 + cells or the percentage of b CD4 + T cells or c CD8 + T cells in CD3 + T cells in splenocytes from mice treated with saline. d Exogenous proBDNF did not alter the percentage of CD3 + T cells in CD45 + cells in splenocytes in septic mice. e–f ProBDNF treatment significantly decreased the percentage of e CD4 + T cells but increased the percentage of f CD8 + T cells in CD3 + T cells in splenocytes in LPS-treated mice. n = 4 in each group. Data were analyzed by one-way ANOVA and followed by Tukey post hoc test, * P

Article Snippet: 4 × 105 cells were put in each well of a 96-well flat-bottom plate and stimulated with 100, 200, or 500 ng ml−1 proBDNF protein (Alomone Labs, Israel, catalog: B243) as introduced by our previous studies [ ], respectively.

Techniques: Mouse Assay, In Vitro, Injection, Isolation, Cell Culture

Journal: Journal of Neuroinflammation

Article Title: ProBDNF promotes sepsis-associated encephalopathy in mice by dampening the immune activity of meningeal CD4+ T cells

doi: 10.1186/s12974-020-01850-0

Figure Lengend Snippet: Increased proBDNF expression in meningeal and peripheral blood immune cells in septic mice. Mice were i.p. injected with LPS (5 mg kg −1 ) and meninges and peripheral blood were harvested for immunofluorescence staining or flow cytometry. a Representative whole mount meningeal immunofluorescence images showed markedly increased proBDNF-positive staining cells in the meninges in mice at 1 day after LPS injection compared to saline injected mice. The high magnification images around the arrows are displayed in a white square as insets. Bar = 100 μm. b – f Representative meningeal single cell flow cytometry images ( upper panel ) and its statistical analysis ( lower panel ) indicated that proBDNF MFI was increased in meningeal b CD3 + T cells, c CD4 + T cells, d CD8 + T cells, and f CD11b + monocytes/macrophages at 1 day after LPS injection. proBDNF in meningeal e CD19 + B cells upregulated until 5 days after LPS injection. n = 10 in the Con group, n = 6 in LPS groups. g – k Upregulation of proBDNF in g CD3 + T cells, h CD4 + T cells, i CD8 + T cells, j CD19 + B cells, and k CD11b + monocytes/macrophages in peripheral blood in LPS-injected mice were detected. n = 9 in Con group, n = 8 in the LPS1d group, n = 4 in the LPS 5d group. Data b – k were analyzed by one-way ANOVA and followed by Tukey post hoc test, * P

Article Snippet: 4 × 105 cells were put in each well of a 96-well flat-bottom plate and stimulated with 100, 200, or 500 ng ml−1 proBDNF protein (Alomone Labs, Israel, catalog: B243) as introduced by our previous studies [ ], respectively.

Techniques: Expressing, Mouse Assay, Injection, Immunofluorescence, Staining, Flow Cytometry

Journal: Journal of Neuroinflammation

Article Title: ProBDNF promotes sepsis-associated encephalopathy in mice by dampening the immune activity of meningeal CD4+ T cells

doi: 10.1186/s12974-020-01850-0

Figure Lengend Snippet: Effect of i.c.v. injection of anti-proBDNF antibody on fear memory and meningeal immune activity in the septic mice. Mice were bilateral i.c.v. injected with 1 μg McAb-proB 3 days before LPS injection. Behavior tests were performed 1 day after LPS injection. a Representative images showed the broad and thorough diffusion of drugs in cerebroventricular lumen following i.c.v. injection with methylene blue. b McAb-proB i.c.v. injection did not influence weight of mice. c–e There was no difference of c fear conditioning acquiring performance in each group, nor McAb-proB increased the freezing time of d contextual or e cued fear conditioning test as compared to IgG control after LPS injection. n = 6 in each group. Data b , c , and e were analyzed by repeated measures ANOVA and followed by Bonferroni post hoc test and data d was analyzed by one-way ANOVA and followed by Tukey post hoc test, * P

Article Snippet: 4 × 105 cells were put in each well of a 96-well flat-bottom plate and stimulated with 100, 200, or 500 ng ml−1 proBDNF protein (Alomone Labs, Israel, catalog: B243) as introduced by our previous studies [ ], respectively.

Techniques: Injection, Activity Assay, Mouse Assay, Diffusion-based Assay

Journal: Journal of Neuroinflammation

Article Title: ProBDNF promotes sepsis-associated encephalopathy in mice by dampening the immune activity of meningeal CD4+ T cells

doi: 10.1186/s12974-020-01850-0

Figure Lengend Snippet: Systemic blockade of proBDNF ameliorated cognitive dysfunction and restored meningeal and peripheral CD4 + T cell ratio in septic mice. Mice were i.p. injected with proBDNF 30 min before LPS (5 mg kg −1 ) injection. Fear conditioning testing was performed 1 day after LPS injection. Meninges and peripheral blood were harvested 5 days after LPS injection for flow cytometry. a McAb-proB did not influence the weight of mice or b fear conditioning acquiring. c , d McAb-proB greatly alleviated memory deficit induced by LPS injection in mice as indicated by the increased freezing time in ( c ) contextual and ( d ) cued fear conditioning tests in the McAb-proB group relative to the IgG control. n = 8 in each group. Data a , b , and d were analyzed by repeated measures ANOVA and followed by Bonferroni post hoc tests. Data c was analyzed by one-way ANOVA and followed by Tukey post hoc test, * P

Article Snippet: 4 × 105 cells were put in each well of a 96-well flat-bottom plate and stimulated with 100, 200, or 500 ng ml−1 proBDNF protein (Alomone Labs, Israel, catalog: B243) as introduced by our previous studies [ ], respectively.

Techniques: Mouse Assay, Injection, Flow Cytometry

Journal: Journal of Neuroinflammation

Article Title: ProBDNF promotes sepsis-associated encephalopathy in mice by dampening the immune activity of meningeal CD4+ T cells

doi: 10.1186/s12974-020-01850-0

Figure Lengend Snippet: Systemic blockade of proBDNF restored meningeal pro-inflammatory microenvironment in septic mice. Mice were i.p. injected with proBDNF 30 min before LPS (5 mg kg −1 ) injection. Meninges were harvested 5 days after LPS injection for qPCR. a The level of CD4 gene expression was higher in the meninges of the McAb-proB group than in IgG controls in septic mice. b–f Gene levels were significantly lower in b IL-1β and c IL-6 but higher in d IL-4, e IFN-γ, and f IL-13 in the meninges after LPS injection in the McAb-proB group as compared to IgG control. n = 5 in each group. All experiments were performed at least in triplicate. Data were analyzed by unpaired T test, * P

Article Snippet: 4 × 105 cells were put in each well of a 96-well flat-bottom plate and stimulated with 100, 200, or 500 ng ml−1 proBDNF protein (Alomone Labs, Israel, catalog: B243) as introduced by our previous studies [ ], respectively.

Techniques: Mouse Assay, Injection, Real-time Polymerase Chain Reaction, Expressing

Journal: Oncogene

Article Title: Neurotensin receptor type 2 protects B-cell chronic lymphocytic leukemia cells from apoptosis

doi: 10.1038/onc.2017.365

Figure Lengend Snippet: NTSR2 phosphorylation in B-CLL and recruitment of G i α proteins. ( a ) Representative western blot analysis of Src phosphorylation in MEC-1 cells overexpressing NTSR2 (pCMV6-NTSR2) in the presence or absence of BDNF (100 ng/ml), pertussis toxin (PTX, 200 ng/ml) or K252a (100 nM) for 24 h. ( b ) Ratio of phosphorylated Src vs pan-Src protein, normalized against actin. Values are means±s.e.m. of three independent experiments in a.u. ( c , d ). After immunoprecipitation (IP) of NTSR2 from MEC-1 cells overexpressing NTSR2 (pCMV6-NTSR2) or not (EV), or from B-CLL cells in the presence or absence of BDNF (100 ng/ml) or NTS (40 μ M /ml), the immunocomplexes were immunoblotted (IB) with anti-G i α1/2 antibodies. ( e , f ) After immunoprecipitation (IP) of anti-pan-phosphoprotein was performed on MEC-1 cells overexpressing NTSR2 and B-CLL cells in the presence or absence of BDNF (100 ng/ml) or SR142948A (67 μ M ), the phosphorylation of NTSR2 was detected by immunoblotting (IB) with anti-NTSR2 antibodies. ( g ) Apoptotic ratio, assessed by cell death ELISA, in B-CLL in the presence or absence of SR142948A (67 μ M ) for 24 h. Values are mean ratios of apoptotic cells (±s.e.m.) of three independent experiments from different patients ( n =3). Significant P -values are indicated in the graphs * P

Article Snippet: Drugs and treatments Cultured cells were incubated for 24 h with 40 μM NTS (Calbiochem/Merck Millipore, Fontenay sous Bois, France) or 100 ng/ml human recombinant BDNF (Alomone labs, Jerusalem, Israel).

Techniques: Western Blot, Immunoprecipitation, Enzyme-linked Immunosorbent Assay

Journal: The Journal of Neuroscience

Article Title: Altered Ion Channels in an Animal Model of Charcot-Marie-Tooth Disease Type IA

doi: 10.1523/JNEUROSCI.3328-04.2005

Figure Lengend Snippet: Kv3.1b is a component of the PNS nodes in Trembler-J nerves. These are images of unfixed teased sciatic nerve fibers from WT and Trembler-J ( TrJ ) mice, labeled as indicated. In WT mice, a few nodes are Kv3.1b positive (apposed arrowheads in A ), whereas

Article Snippet: Frozen sections and teased fibers were permeabilized by immersion in -20°C acetone for 10 min, blocked at room temperature for 1 h with 5% fish skin gelatin containing 0.1% Triton X-100 in PBS, and incubated overnight at 4°C with various combinations of primary antibodies: rabbit antisera against KCNQ2 [1:200 ( )], Kv3.1b (1:100; Alomone Labs, Jerusalem, Israel), Kv1.2 (1:100; Alomone Labs), Nav1.6 [1:100 ( )], Nav1.8 [1:100 ( )], Nav1.8 [1:500 ( )], ankyrin-G [1:100 ( ; )], contactin [1:100 ( )], ezrin-binding protein 50 kDa (EBP-50) (ab3452; 1:100; Abcam, Cambridge, MA), syndecan-3 [1:300 ( )], or Caspr [1:500 ( )]; and mouse monoclonal antibodies against panNav channels (K58/35; 1:250; Sigma, St. Louis, MO), Nav1.2 [1:100 ( ; )], Nav1.2 (1:50; Upstate Biotechnology, Lake Placid, NY), Caspr [1:50 ( )], annexin II light chain (1:50; BD Transduction Laboratories, San Jose, CA), or myelin-associated glycoprotein (MAG) (clone 513, 1:100; Boehringer Mannheim, Indianapolis, IN); rat monoclonal antibody against E-cadherin (1:50; Sigma); chicken antibody against βIV-spectrin [1:500 ( )]; and a soluble construct of receptor protein tyrosine phosphatase β conjugated to the human Fc fragment (RPTPβ-Fc) [1:3 ( )].

Techniques: Mouse Assay, Labeling