Journal: Journal of Perinatal Medicine

Article Title: Nicotine affects the expression of brain-derived neurotrophic factor mRNA and protein in the hippocampus of hypoxic newborn piglets

doi: 10.1515/jpm.2009.081

Figure Lengend Snippet: Figure 1 Relative expression of BDNF mRNA in hippocampus. The group treated with nicotine 130 mg/kg/h show a significant increase in BDNF mRNA expression compared to the group treated with saline. 1snicotine 130 mg/kg/h; 2snicotine 260 mg/kg/h; 3sadrena- line 0.05 mg/kg/minqsaline; 4ssaline. *Ps0.038 for nicotine 130 mg/kg/h vs. saline.

Article Snippet: Monoclonal anti-human BDNF antibody titer 1:25 (Clone 35928; R&D Systems, Minneapolis, MN, USA) was used as primary antibody.

Techniques: Expressing, Saline

Journal: Journal of Perinatal Medicine

Article Title: Nicotine affects the expression of brain-derived neurotrophic factor mRNA and protein in the hippocampus of hypoxic newborn piglets

doi: 10.1515/jpm.2009.081

Figure Lengend Snippet: Figure 4 Immunohistochemistry of BDNF protein, nicotine vs. saline. Pictures from the dentate gyrus 20=enlarged. A is from an ani- mal treated with nicotine 130 mg/kg/h; B is from an animal treat- ed with saline; picture A presents a larger number of BDNF positive cells than picture B.

Article Snippet: Monoclonal anti-human BDNF antibody titer 1:25 (Clone 35928; R&D Systems, Minneapolis, MN, USA) was used as primary antibody.

Techniques: Immunohistochemistry, Saline

Journal: Journal of Perinatal Medicine

Article Title: Nicotine affects the expression of brain-derived neurotrophic factor mRNA and protein in the hippocampus of hypoxic newborn piglets

doi: 10.1515/jpm.2009.081

Figure Lengend Snippet: Figure 2 Box plot of % BDNF positive cells in the dentate gyrus of the hippocampus. There are significantly more BDNF positive cells in the dentate gyrus of the animals treated with nicotine 130 mg/kg/h com- pared to the saline treated animals. *Ps0.009.

Article Snippet: Monoclonal anti-human BDNF antibody titer 1:25 (Clone 35928; R&D Systems, Minneapolis, MN, USA) was used as primary antibody.

Techniques: Saline

Journal: Journal of Perinatal Medicine

Article Title: Nicotine affects the expression of brain-derived neurotrophic factor mRNA and protein in the hippocampus of hypoxic newborn piglets

doi: 10.1515/jpm.2009.081

Figure Lengend Snippet: Figure 3 Immunohistochemistry of BDNF protein. Picture from the dentate gyrus 40=enlarged. The cells with the large, dark brown nucleuses are positive (black arrows), the cells with light brown or blue nucleuses are negative (open arrows).

Article Snippet: Monoclonal anti-human BDNF antibody titer 1:25 (Clone 35928; R&D Systems, Minneapolis, MN, USA) was used as primary antibody.

Techniques: Immunohistochemistry

Journal: Advanced science (Weinheim, Baden-Wurttemberg, Germany)

Article Title: Locus Coeruleus-Dorsolateral Septum Projections Modulate Depression-Like Behaviors via BDNF But Not Norepinephrine.

doi: 10.1002/advs.202303503

Figure Lengend Snippet: Figure 7. BDNF was necessary for the LCTH-dLS circuit to sustain the antidepressant-like effects of S-ketamine but not fluoxetine. A) Schematic rep- resentation of the Cre-dependent AAV expressing BDNF-shRNA-EGFP in the LCTH-dLS circuit. B) Timeline of experiments. C) Representative social interaction tracks of BDNF-shRNA and Con-shRNA mice and the time spent in the interaction zone in the SIT in different groups (n = 9). D) Locomotor activity and depression-like behaviors in BDNF-shRNA and control-shRNA mice injected with saline or S-ketamine (n = 9). E) Schematic representation of virus injection. F) Timeline of the experiments. G) Representative social interaction tracks of BDNF-shRNA and Con-shRNA mice and the time spent in the interaction zone in the SIT in different groups (n = 8). H) Locomotor activity and depression-like behaviors in BDNF-shRNA and Con-shRNA mice injected with saline or fluoxetine (n = 8). Data represented as mean ± SEM. *p < 0.05, **p < 0.01, ***p < 0.001; n.s., not significant. Two-way ANOVA followed by Bonferroni’s post hoc analysis for (C,D) and (G,H). The statistical details can be found in Table S1, Supporting Information.

Article Snippet: Drugs were prepared: CNO (1 mg kg−1, i.p., diluted in 0.9% saline; 3 mM, 1 μL per site, CAS NO.: C0832; Sigma-Aldrich), K252a (1 μM, 1 μL per site, CAS NO.:99533-80-9, Sigma-Aldrich), propranolol (5 mM, 1 μL per site, CAS NO.:318-98-9, Sigma-Aldrich), phentolamine (10 mM, 1 μL per site, CAS NO.: 73-05-2, Sigma-Aldrich), Recombinant Human BDNF Protein (200 ng in 0.9% saline for local injection, 1 μL per site, CAS NO.:248-BDB; R&D Systems), anti-BDNF neutralizing antibody (0.2 μg in 0.9% saline for local injection, 0.2 μL per site, EMD Millipore), Sketamine (10 mg kg−1, i.p., diluted in 0.9% saline, CAS NO.:1430202-702, Sigma-Aldrich), R-ketamine (10 mg kg−1, i.p., diluted in 0.9% saline, CAS NO.: 1430202-69-9, Sigma-Aldrich), and fluoxetine (20 mg kg−1 i.p., diluted in 0.9% saline, CAS NO.:56296-78-7, Sigma-Aldrich).

Techniques: Expressing, shRNA, Activity Assay, Control, Injection, Saline, Virus

Journal: Journal of neurochemistry

Article Title: Chronic cortisol exposure promotes the development of a GABAergic phenotype in the primate hippocampus.

doi: 10.1111/j.1471-4159.2004.02760.x

Figure Lengend Snippet: Fig. 5 Chronic cortisol exposure increases BDNF immunostaining, but not BDNF mRNA expression, in the primate hippocampus. (a, c) Representative low-power photomicrographs (2X) depicting the pat- tern of BDNF immunostaining and mRNA expression, respectively, in the primate hippocampus. (b) Cortisol significantly increases BDNF immunostaining in the dentate polymorphic layer and CA3 mossy fibers. (d) Cortisol has no effect on BDNF mRNA expression. *p ¼ 0.05; DPL, dentate polymorphic layer; DGL, dentate granule layer; CA-3 MF, CA3 mossy fiber projections.

Article Snippet: The following primary antibodies were used: monoclonal anti-CaB-D (1 : 1600; Sigma, St Louis, MO, USA), monoclonal anti-PV (1 : 800; Sigma), polyclonal anti-CR (1 : 1600; Sigma), polyclonal anti-GAD65/67 (1 : 56 000; Chemicon, Temecula, CA, USA), and monoclonal antihuman BDNF (1 : 200; R&D Systems, Minneapolis, MN, USA).

Techniques: Immunostaining, Expressing

Journal: Journal of Virology

Article Title: Antibody to Varicella-Zoster Virus Immediate-Early Protein 62 Augments Allodynia in Zoster via Brain-Derived Neurotrophic Factor

doi: 10.1128/jvi.02061-09

Figure Lengend Snippet: FIG. 1. Reactivity of anti-IE62 and anti-BDNF MAbs to IE62 and BDNF. (A) Schematic presentation of IE62 GST fusion proteins from GST-1 to GST-5 and GST-A to GST-G, comprising the entire IE62 molecule. (B) VZV IE62 proteins were expressed as GST fusion proteins using vector pGEX-4T-1 and stained with Coomassie brilliant blue R-250 (CBB). (C) An SDS-PAGE gel of IE62-GST fusion proteins prepared as described for panel B was probed with anti-IE62 and anti-BDNF MAbs, and both recognized IE62 GST-2, GST-C, and GST-F by Western blot analysis. (D) BDNF (R&D and Sigma) was detected as a monomer by CBB staining, as shown in the left panel, and anti-IE62 (2-b) and anti-BDNF MAbs reacted strongly to the BDNF dimer and weakly to the monomer, as shown in the right panel. (E) Immunofluorescence was performed in A549 cells infected with the Kawaguchi strain of VZV 4 days after cell-free virus infection. The plaque was surrounded by the uninfected cells at low magnification. Anti-IE62 (2-b) and anti-BDNF MAbs stained the nuclei of VZV-infected cells similarly without the staining of uninfected cells. Scale bar, 50 m.

Article Snippet: After being blocked with skim milk, the membranes were incubated with anti-IE62 MAb (1:10,000 dilution of ascites) or anti-human BDNF monoclonal antibody (0.5- g/ml dilution; R&D Systems, Inc., Minneapolis, MN) at room temperature for 3 h. After a wash with phosphate-buffered saline (PBS), the membranes were incubated with peroxidase-conjugated goat anti-mouse immunoglobulin G (H L) [IgG(H L)] (1: 2,000 dilution; Jackson ImmunoResearch Laboratories, Inc., West Grove, PA) for 1 h, and then they were developed by the electrogenerated chemiluminescence (ECL) method (Nacalai Tesque, Kyoto, Japan).

Techniques: Plasmid Preparation, Staining, SDS Page, Western Blot, Infection, Virus

Journal: Journal of Virology

Article Title: Antibody to Varicella-Zoster Virus Immediate-Early Protein 62 Augments Allodynia in Zoster via Brain-Derived Neurotrophic Factor

doi: 10.1128/jvi.02061-09

Figure Lengend Snippet: FIG. 2. Determination of epitope of IE62 recognized by anti-IE62 and anti-BDNF-MAbs. (A) Locations of peptides used for determination of anti-IE62 MAb. Anti-IE62 and anti-BDNF MAbs recognized IE62 GST-2, GST-C, and GST-F, as shown in Fig. 1, and the overlapping region was amino acids 414 to 447 of IE62. This region was covered by three peptides (amino acids 414 to 459) to block the interaction with MAbs. (B) IE62-GST-F was blotted and probed by anti-BDNF and anti-IE62 MAbs in the absence and presence of 180 g/ml of each peptide. Lanes 1 and 2 indicate the amounts of IE62-GST-F at 220 and 88 ng/lane, respectively. IE62-GST-F is indicated by the arrows at a molecular mass of 37.2 kDa. p414-429 blocked the interaction of both anti-IE62 and anti-BDNF MAbs, but the other two peptides failed to block. The Western blotting sheets treated with each peptide were collected and separated by the line. (C) BDNF was blotted and probed by anti-BDNF and anti-IE62 MAbs. Lanes 1 and 2 indicate BDNF expressed in Escherichia coli (Sigma) at 160 ng/lane and expressed in insect cells (R&D Systems) at 260 ng/lane, respectively. BDNF monomers and dimers are indicated by the arrows at 13.6 and 27.5 kDa, respectively. Three peptides failed to inhibit interactions of the monomer and dimer of BDNF with anti-IE62 and anti-BDNF MAbs under the same conditions. Anti-IE62 and anti-BDNF MAbs recognized the linear epitope (p414-429) of IE62. Both MAbs may prefer the recognition of the conformational epitope of BDNF, particularly the epitope formed by dimeric BDNF, rather than a linear epitope.

Article Snippet: After being blocked with skim milk, the membranes were incubated with anti-IE62 MAb (1:10,000 dilution of ascites) or anti-human BDNF monoclonal antibody (0.5- g/ml dilution; R&D Systems, Inc., Minneapolis, MN) at room temperature for 3 h. After a wash with phosphate-buffered saline (PBS), the membranes were incubated with peroxidase-conjugated goat anti-mouse immunoglobulin G (H L) [IgG(H L)] (1: 2,000 dilution; Jackson ImmunoResearch Laboratories, Inc., West Grove, PA) for 1 h, and then they were developed by the electrogenerated chemiluminescence (ECL) method (Nacalai Tesque, Kyoto, Japan).

Techniques: Blocking Assay, Western Blot

Journal: Journal of Virology

Article Title: Antibody to Varicella-Zoster Virus Immediate-Early Protein 62 Augments Allodynia in Zoster via Brain-Derived Neurotrophic Factor

doi: 10.1128/jvi.02061-09

Figure Lengend Snippet: FIG. 4. Reactivity of sera from patients with zoster and PHN to IE62 and BDNF and augmentation of BDNF activity. (A) The table summarizes the results of patient antibodies to IE62-2 and the BDNF dimer; the age and sex (male, M; female, F) are indicated. ND indi- cates not detected, and 1, 2, and 3 indicate the detection levels in Western blotting. (B) Antibodies to IE62-2 and the BDNF dimer are shown as representative patterns in the blot. (C) Four of the five patient sera in the table significantly enhanced the transcription of BDNF at the final dilution of 1:10,000. The amounts of BDNF tran- scripts in the neurons treated with BDNF alone and with BDNF and serum are represented as the means standard errors of the means (n 4). * and ** indicate P 0.05 and P 0.01, respectively, compared to BDNF alone (Student’s t test).

Article Snippet: After being blocked with skim milk, the membranes were incubated with anti-IE62 MAb (1:10,000 dilution of ascites) or anti-human BDNF monoclonal antibody (0.5- g/ml dilution; R&D Systems, Inc., Minneapolis, MN) at room temperature for 3 h. After a wash with phosphate-buffered saline (PBS), the membranes were incubated with peroxidase-conjugated goat anti-mouse immunoglobulin G (H L) [IgG(H L)] (1: 2,000 dilution; Jackson ImmunoResearch Laboratories, Inc., West Grove, PA) for 1 h, and then they were developed by the electrogenerated chemiluminescence (ECL) method (Nacalai Tesque, Kyoto, Japan).

Techniques: Activity Assay, Western Blot