Journal: International Journal of Molecular Sciences

Article Title: Serotonin 5-HT 4 Receptor Agonists Improve Facilitation of Contextual Fear Extinction in an MPTP-Induced Mouse Model of Parkinson’s Disease

doi: 10.3390/ijms20215340

Figure Lengend Snippet: Analysis of the number of p-CREB-positive cells in the dentate gyrus (DG) of the hippocampus after extinction training. ( A ) Immunohistochemistry for p-CREB (red) and NeuN (green) in the DG after fear extinction. (a) Control + vehicle, (b) MPTP + vehicle, (c) MPTP + prucalopride 3.0 mg/kg, (d) MPTP + velusetrag 3.0 mg/kg. Scale bar = 100 µm. The images of “a-high”, “b-high”, “c-high”, and “d-high” were partially expanded from the images of the white dotted box area in a, b, c, and d, respectively. Some p-CREB-positive cells co-localize with NeuN (yellowish red, indicated by white arrows). ( B ) The number of p-CREB-positive cells in the DG. The mean was calculated by averaging the number of p-CREB-positive cells counted from three 50-µm sections (obtained from a position 1.82–2.02 mm posterior to the bregma). Data are expressed as the mean ± SEM; n = 3–4 per group. ** p < 0.01 vs. control + vehicle, ++ p < 0.01 vs. MPTP + vehicle (one-way ANOVA followed by Tukey’s post-hoc test).

Article Snippet: The sections were then permeabilized with 0.5% (v/v) Triton X-100 in PBS (T-PBS) for 1 h, blocked in goat serum solution (1:100 in T-PBS) for 1 h, and incubated for 48 h at 4 °C in rabbit anti-phosphorylated CREB (p-CREB) polyclonal antibody (1:800 in T-PBS; Cell Signaling Technology Japan, KK., Tokyo, Japan).

Techniques: Immunohistochemistry, Control

Journal: Scientific Reports

Article Title: Early neurotrophic pharmacotherapy rescues developmental delay and Alzheimer’s-like memory deficits in the Ts65Dn mouse model of Down syndrome

doi: 10.1038/srep45561

Figure Lengend Snippet: ( a – i ) Western blot analyses of synaptophysin, PSD95, CNTF, BDNF, TrkB, pGSK3β (pSer9)/GSK3β, pCREB (pSer133)/CREB, and PKAcα in the hippocampus of 3-week-old mice. P021 treatment ameliorated deficits in synaptophysin, BDNF, and PKAcα expression, and increased pGSK3β/GSK3β and pCREB/CREB ratios. ( a ) Representative Western blots. ( b – i ) Quantification of the Western blots. ( j – m ) The mRNA levels of synaptophysin, BDNF, TrkB, and CREB in the cortex. P021 treatment significantly enhanced synaptophysin and BDNF mRNA levels. The data are presented as mean ± S.E.M. from WT-V-WT (n = 6), WT-P-WT (n = 7), Dn-V-WT (n = 7), Dn-P-WT (n = 7), Dn-V-Dn (n = 7), and Dn-P-Dn (n = 6) mice. * p < 0.05; ** p < 0.01; *** p < 0.001. One-way ANOVA with Bonferroni’s post - hoc test.

Article Snippet: The following primary antibodies were used: mouse monoclonal anti-synaptophysin (1:3000, Millipore, Temecula, CA, USA), rabbit monoclonal anti-PSD95 (1:1000, Cell Signaling Technology, Danvers, MA, USA), rabbit polyclonal anti-BDNF, N-20 (1:500, Santa Cruz Biotechnology, Santa Cruz, CA, USA), rabbit polyclonal anti-CNTF, FL-200 (1:500, Santa Cruz Biotechnology, Santa Cruz, CA, USA), rabbit polyclonal anti-TrkB (total) (1∶500; Santa Cruz Biotechnology, Santa Cruz, CA, USA), rabbit polyclonal anti-CREB (1:1000, Cell Signaling Technology, Danvers, MA, USA), rabbit polyclonal anti-phosphorylated CREB, Ser133 (1:1000, Cell Signaling Technology, Danvers, MA, USA), rabbit monoclonal anti GSK3β (1:1000, Cell Signaling, Danvers, MA, USA), rabbit polyclonal anti-phosphorylated GSK3β, Ser9 (1:1000, Cell Signaling, Danvers, MA, USA), rabbit polyclonal anti-protein kinase A catalytic subunit alpha (PKAcα) (1:1000, Santa Cruz Biotechnology, Santa Cruz, CA, USA), mouse monoclonal anti-STAT3 (1:1000; Cell Signaling Technology, Danvers, MA, USA), rabbit monoclonal anti-pSTAT3, Tyr705 (1:1000; Cell Signaling Technology, Danvers, MA, USA), and rabbit polyclonal antibody to GAPDH (1:1000, Santa Cruz Biotechnology, Santa Cruz, CA, USA) as loading control.

Techniques: Western Blot, Expressing

Journal: Scientific Reports

Article Title: Early neurotrophic pharmacotherapy rescues developmental delay and Alzheimer’s-like memory deficits in the Ts65Dn mouse model of Down syndrome

doi: 10.1038/srep45561

Figure Lengend Snippet: ( a – i ) Western blot analyses of synaptophysin, PSD95, CNTF, BDNF, TrkB, pGSK3β (pSer9)/GSK3β, pCREB(pSer133)/CREB, and PKAcα in the hippocampus of ~7-month-old mice. P021 treatment ameliorated deficits in synaptophysin, BDNF, and PKAcα expression, and increased pGSK3β/GSK3β and pCREB/CREB ratios. ( a ) Representative Western blots. ( b – i ) Quantification of the Western blots. ( j – m ) The mRNA levels of synaptophysin, BDNF, TrkB, and CREB in the cortex. P021 treatment ameliorated BDNF mRNA level deficit. The data are presented as mean ± S.E.M. from WT-V-WT (n = 7), WT-P-WT (n = 7), Dn-V-WT (n = 6), Dn-P-WT (n = 6), Dn-V-Dn (n = 7), and Dn-P-Dn (n = 7) mice. * p < 0.05; ** p < 0.01; *** p < 0.001. One-way ANOVA with Bonferroni’s post - hoc test.

Article Snippet: The following primary antibodies were used: mouse monoclonal anti-synaptophysin (1:3000, Millipore, Temecula, CA, USA), rabbit monoclonal anti-PSD95 (1:1000, Cell Signaling Technology, Danvers, MA, USA), rabbit polyclonal anti-BDNF, N-20 (1:500, Santa Cruz Biotechnology, Santa Cruz, CA, USA), rabbit polyclonal anti-CNTF, FL-200 (1:500, Santa Cruz Biotechnology, Santa Cruz, CA, USA), rabbit polyclonal anti-TrkB (total) (1∶500; Santa Cruz Biotechnology, Santa Cruz, CA, USA), rabbit polyclonal anti-CREB (1:1000, Cell Signaling Technology, Danvers, MA, USA), rabbit polyclonal anti-phosphorylated CREB, Ser133 (1:1000, Cell Signaling Technology, Danvers, MA, USA), rabbit monoclonal anti GSK3β (1:1000, Cell Signaling, Danvers, MA, USA), rabbit polyclonal anti-phosphorylated GSK3β, Ser9 (1:1000, Cell Signaling, Danvers, MA, USA), rabbit polyclonal anti-protein kinase A catalytic subunit alpha (PKAcα) (1:1000, Santa Cruz Biotechnology, Santa Cruz, CA, USA), mouse monoclonal anti-STAT3 (1:1000; Cell Signaling Technology, Danvers, MA, USA), rabbit monoclonal anti-pSTAT3, Tyr705 (1:1000; Cell Signaling Technology, Danvers, MA, USA), and rabbit polyclonal antibody to GAPDH (1:1000, Santa Cruz Biotechnology, Santa Cruz, CA, USA) as loading control.

Techniques: Western Blot, Expressing