Journal: PLoS ONE

Article Title: Neurogenic and Neurotrophic Effects of BDNF Peptides in Mouse Hippocampal Primary Neuronal Cell Cultures

doi: 10.1371/journal.pone.0053596

Figure Lengend Snippet: (A–C) Peptide B-5 induces the expression of the neuronal markers β-III-tubulin, MAP2, NFM, and NeuN in E18 primary hippocampal cell cultures. Representative confocal images illustrating double immunolabeling of β–III-tubulin (green) and MAP2 (red) and NFM (green) and NeuN (red) in cells treated for 5 days with (A) Peptide B-5 at 1 µM, (B) with BDNF, 20 ng/ml (0.79 nM) for 5 days, and,(C) vehicle only (control, C). TOPRO-3 (blue) was used to stain the nuclei. Magnification bar = 10 µm. The bottom panel shows the level of fluorescence intensity using Image J software to measure the level of expression. (D, E) BDNF peptides (B-5, B-4, and B-3) induce the expression of neuronal markers in E18 primary hippocampal neurons. (D) Representative Western blots showing an increase in the expression of the neuronal markers PSD95, NeuN, NFM, and MAP-2 in cells treated with the peptides (B5, B-4 and B-3) at concentrations of 0.1 µM and 1 µM or BDNF at concentrations of 20 or 100 ng/ml (0.79 or 3.95 nM respectively). GPADH was used as a loading control. (E) Quantification of the Western blots of neuronal markers shown in D. The integrated density value of the bands in Western blots was determined using densitometry (Fuji software, Multi Gauge, Version 3.0), and data was normalized to GAPDH and to control (medium treated cells for 5 days, C 5d). Data are shown as mean ± standard deviation, n = 3. 10% SDS-PAGE gels. * p <0.05, ** p <0.01, *** p <0.001. One-way ANOVA/post-hoc test/Student’s t test.

Article Snippet: The following primary antibodies were used: rabbit polyclonal anti-NFM (early neuronal marker, 1∶1000; Chemicon/Millipore, Billerica, MA, USA); mouse monoclonal anti-NeuN, clone A60 (late neuronal marker, 1∶500; Millipore, Billerica, MA, USA); rabbit polyclonal anti-Tuj-1, β–III-tubulin (early neuronal marker, Covance; 1∶800, Emeryville, CA, USA); mouse monoclonal SMI 52 to MAP-2 (dendritic marker, 1∶1000; Covance, Emeryvilly, CA, USA); rabbit polyclonal anti-Synapsin I (pre-synaptic marker, 1∶2000; Stressgen, Farmingdale, NY, USA); rabbit monoclonal anti- PSD95 (post-synaptic marker, 1∶1000; Cell signaling, Danvers, MA, USA); anti tau 92e (axonal marker, 1∶10000; , ; rabbit polyclonal anti- BDNF (N-20) (1∶200 Santa Cruz Biotechnology, Santa Cruz, CA, USA ); rabbit polyclonal anti-TrkB (total) (1∶500; Santa Cruz Biotechnology, Santa Cruz, CA, USA); rabbit polyclonal anti- pTrkB (Tyr706) (1∶400; Santa Cruz Biotechnology, Santa Cruz, CA, USA), TrkC (1∶1000; Cruz Biotechnology, Santa Cruz, CA, USA); and pTrkY490 (1∶500; Upstate USA Inc., Charlottesville, VA, USA).

Techniques: Expressing, Immunolabeling, Control, Staining, Fluorescence, Software, Western Blot, Standard Deviation, SDS Page

Journal: British Journal of Nutrition

Article Title: Memory-enhancing effect of Mori Fructus via induction of nerve growth factor

doi: 10.1017/s0007114512004710

Figure Lengend Snippet: Fig. 3. Effect of Mori Fructus extract (ME) on pre- and post-synaptic formation and acetylcholine synthesisation in the mouse hippocampus. The synaptophysin (SYN), post-synaptic density-95 (PSD95) and choline acetyltransferase (ChAT) proteins in the hippocampal tissue were expressed using Western blotting after the administration of ME (20, 100 and 500 mg/kg per d, per os) for 7 d. The graphs display densitometric analyses of the expression ratios of (a) SYN:b-actin, (b) PSD95:b-actin and (c) ChAT:b-actin. Values are means, with their standard errors represented by vertical bars. Mean value was significantly different from that of the control group: * P,0·05, ** P,0·01, *** P,0·001.

Article Snippet: Rat monoclonal anti-5-bromo-2-deoxyuridine (BrdU), rabbit monoclonal anti-phosphorylated extracellular signal-regulated kinase (pERK), rabbit monoclonal anti-phosphorylated cyclic AMP response element-binding protein (pCREB), rabbit monoclonal anti-ERK, rabbit monoclonal anti-CREB and goat polyclonal anti-doublecortin (DCX) were purchased from Santa Cruz Biotechnology, Inc. Rabbit polyclonal anti-post-synaptic density-95 (PSD95) was purchased from Abcam.

Techniques: Western Blot, Expressing, Control

Journal:

Article Title: Architecture of cannabinoid signaling in mouse retina

doi: 10.1002/cne.22429

Figure Lengend Snippet: Antibodies used in this study.

Article Snippet: PSD95 , Purified recombinant rat PSD-95 (Post Synaptic Density 95 kDa) , Genetex, Irvine, CA, #GTX80682, mouse monoclonal , 1:1000.

Techniques: Purification, Affinity Purification, Recombinant