α btx  (Alomone Labs)

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: 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: American Journal of Physiology - Regulatory, Integrative and Comparative Physiology

Article Title: Dietary sodium modulates the interaction between efferent renal sympathetic nerve activity and afferent renal nerve activity: role of endothelin

doi: 10.1152/ajpregu.91029.2008

Figure Lengend Snippet: HNa diet, in vivo: effects of an ETB-R antagonist on the ARNA responses to reflex increases in ERSNA.

Article Snippet: After completion of the protocol for TSA for detection of GFAP or CGRP, the tissue sections were incubated with antiserum for ETB-R (rabbit; 1:100, Alomone Labs) and processed by the indirect immunofluorescence technique.

Techniques: In Vivo

Journal: American Journal of Physiology - Regulatory, Integrative and Comparative Physiology

Article Title: Dietary sodium modulates the interaction between efferent renal sympathetic nerve activity and afferent renal nerve activity: role of endothelin

doi: 10.1152/ajpregu.91029.2008

Figure Lengend Snippet: Immunofluorescence double-labeling of T 13 DRG shows colocalization of ETB-R-like immunoreactivity (ETB-R-LI) with glial fibrillary acidic protein (GFAP)-LI (arrows) surrounding neuronal cell bodies ( a–c ). Also, many ETB-R-ir fiber-like structures

Article Snippet: After completion of the protocol for TSA for detection of GFAP or CGRP, the tissue sections were incubated with antiserum for ETB-R (rabbit; 1:100, Alomone Labs) and processed by the indirect immunofluorescence technique.

Techniques: Immunofluorescence, Labeling

Journal: American Journal of Physiology - Regulatory, Integrative and Comparative Physiology

Article Title: Dietary sodium modulates the interaction between efferent renal sympathetic nerve activity and afferent renal nerve activity: role of endothelin

doi: 10.1152/ajpregu.91029.2008

Figure Lengend Snippet: Our studies suggest that ERSNA-induced increases in NE release lead to an increase in renal pelvic PGE 2 synthesis/release. In conditions of HNa dietary intake, activation of ETB-R on or close to unmyelinated Schwann cells surrounding the sensory nerves

Article Snippet: After completion of the protocol for TSA for detection of GFAP or CGRP, the tissue sections were incubated with antiserum for ETB-R (rabbit; 1:100, Alomone Labs) and processed by the indirect immunofluorescence technique.

Techniques: Activation Assay

Journal: American Journal of Physiology - Regulatory, Integrative and Comparative Physiology

Article Title: Dietary sodium modulates the interaction between efferent renal sympathetic nerve activity and afferent renal nerve activity: role of endothelin

doi: 10.1152/ajpregu.91029.2008

Figure Lengend Snippet: Immunofluorescence for ETB-R ( a ) in renal pelvic tissue shows nerve fiber-like structures (arrows) among smooth muscle cells. ETA-R-LI is present on smooth muscle cells in the renal pelvic wall and adjacent blood vessels (arrows, b ).

Article Snippet: After completion of the protocol for TSA for detection of GFAP or CGRP, the tissue sections were incubated with antiserum for ETB-R (rabbit; 1:100, Alomone Labs) and processed by the indirect immunofluorescence technique.

Techniques: Immunofluorescence

Journal: American Journal of Physiology - Regulatory, Integrative and Comparative Physiology

Article Title: Dietary sodium modulates the interaction between efferent renal sympathetic nerve activity and afferent renal nerve activity: role of endothelin

doi: 10.1152/ajpregu.91029.2008

Figure Lengend Snippet: Protein expression of ETB-R ( left ) and ETA-R ( right ) in T 9 –L 1 DRG and renal pelvic wall from rats fed the HNa and LNa diet. The ratio of ET-R/actin protein expression was normalized to the average value of the ratio in HNa diet rats. *

Article Snippet: After completion of the protocol for TSA for detection of GFAP or CGRP, the tissue sections were incubated with antiserum for ETB-R (rabbit; 1:100, Alomone Labs) and processed by the indirect immunofluorescence technique.

Techniques: Expressing

Journal: American Journal of Physiology - Regulatory, Integrative and Comparative Physiology

Article Title: Dietary sodium modulates the interaction between efferent renal sympathetic nerve activity and afferent renal nerve activity: role of endothelin

doi: 10.1152/ajpregu.91029.2008

Figure Lengend Snippet: A : in vivo, effects of renal pelvic administration of the ETB-R antagonist BQ788 on the ERSNA and ARNA responses to placing the rat's tail in 47°C water in rats fed HNa diet. B : effects of renal pelvic administration of the ETA-R antagonist BQ123

Article Snippet: After completion of the protocol for TSA for detection of GFAP or CGRP, the tissue sections were incubated with antiserum for ETB-R (rabbit; 1:100, Alomone Labs) and processed by the indirect immunofluorescence technique.

Techniques: In Vivo

Journal: American Journal of Physiology - Regulatory, Integrative and Comparative Physiology

Article Title: Dietary sodium modulates the interaction between efferent renal sympathetic nerve activity and afferent renal nerve activity: role of endothelin

doi: 10.1152/ajpregu.91029.2008

Figure Lengend Snippet: Immunofluorescence double-labeling of renal pelvic tissue shows ETB-R-ir fiber-like structures among smooth muscle cells ( a–c ) close to calcitonin gene-related peptide-immunoreactive (CGRP-ir) sensory nerve fibers ( d–f ). αSMA,

Article Snippet: After completion of the protocol for TSA for detection of GFAP or CGRP, the tissue sections were incubated with antiserum for ETB-R (rabbit; 1:100, Alomone Labs) and processed by the indirect immunofluorescence technique.

Techniques: Immunofluorescence, Labeling

Journal: eLife

Article Title: Synchronous and asynchronous modes of synaptic transmission utilize different calcium sources

doi: 10.7554/eLife.01206

Figure Lengend Snippet: Delayed release in ω-conotoxin GVIA-treated fish is also observed by means of the exocytotic indicator synaptopHluorin. ( A ) Images taken from a single focal plane of the CaP motor neuron terminals showing the motor neuron fill with Alexa Fluor 647 (gray), postsynaptic labeling with α-btx (red), peak stimulus-induced synaptopHluorin fluorescence (green) and α-btx/synaptopHluorin overlay. These images are shown for a single control (left) and ω-conotoxin GVIA-treated (right) fish. The scale bar corresponds to 10 μm. ( B ) Stimulus-driven fluorescence change, expressed as ΔF/F 0 for the three representative boutons shown for control (gray) and ω-conotoxin GVIA treated (colored) synaptopHluorin motor neurons. The synaptopHluorin signal was measured at each ROI during the time course of 100 Hz stimulation (indicated by the black bar in B ). ( C ) The histogram showing time required to reach 50% maximal fluorescence increase for boutons of control (gray, 127 boutons from 8 fish) and ω-conotoxin GVIA-treated fish (red, 55 boutons from 4 fish). Experiments were performed with 5 mM EGTA in the intracellular solution. DOI: http://dx.doi.org/10.7554/eLife.01206.005

Article Snippet: TTX, ω-conotoxin GVIA and α-btx were obtained from Alomone Labs (Jerusalem, Israel).

Techniques: Fluorescence In Situ Hybridization, Labeling, Fluorescence

Journal: eLife

Article Title: Synchronous and asynchronous modes of synaptic transmission utilize different calcium sources

doi: 10.7554/eLife.01206

Figure Lengend Snippet: Stimulus-driven calcium signals in CaP motor neuron terminals occurred at synaptic boutons. The fill corresponds to a maximal intensity projection image of the motor neuron filled with Alexa Fluor 647. An arrowhead indicates the soma. A single plane of focus in the filled neuron showing postsynaptic α-btx label, peak Fluo-4 calcium signal and merged α-btx and Fluo-4 signal. The scale bar corresponds to 10 μm. DOI: http://dx.doi.org/10.7554/eLife.01206.006

Article Snippet: TTX, ω-conotoxin GVIA and α-btx were obtained from Alomone Labs (Jerusalem, Israel).

Techniques: