Journal: Molecular medicine reports

Article Title: Naringin ameliorates memory deficits and exerts neuroprotective effects in a mouse model of Alzheimer's disease by regulating multiple metabolic pathways.

doi: 10.3892/mmr.2021.11971

Figure Lengend Snippet: Figure 8. Naringin exerts neuroprotective effects by regulating the glutamate receptor system and apoptosis. (A) Protein levels of NMDAR1, GluR2 and CAMKII. (B) Protein levels of Bad, Bcl‑2 and cleaved‑caspase‑3. **P<0.01 vs. sham group; #P<0.05 and ##P<0.01 vs. model group; &P<0.05, &&P<0.01 vs. naringin group. NMDAR1, glutamate receptor subunit 1; GluR2, glutamate receptor 2; CAMKII, calcium/calmodulin‑dependent protein kinase type II.

Article Snippet: After blocking with 5% non‐fat dried milk for 2.5 h at 37 ̊C, the PVDF membrane was incubated with primary antibodies against: APP (1:1,000; cat. no. bs‐12503R; BIOSS), BACE1 (1:1,000; cat. no. 5606T; Cell Signaling Technology, Inc.), CDK5 (1:1,000; cat. no. bs‐10258Rm; BIOSS), p‐Tau396 (1:1,000; cat. no. bs‐3446R; BIOSS), glutamate receptor subunit 1 (NMDAR1) (1:1,000; cat. no. bs‐23343R; BIOSS), glutamate receptor 2 (GluR2; 1:1,000; cat. no. bs‐13385R; BIOSS), calcium/calmod‐ ulin‐dependent protein kinase type II (CAMKII; 1:1,000; Boster), Bad (1:500; cat. no. A00183; Boster), caspase‐3 (1:500; cat. no. bs‐0081R; BIOSS), Bcl‐2 (1:500; cat. no. bs‐0032R; BIOSS), ERβ (1:1,000; cat. no. kl437Hu22; KALANG; https://www.biomart.cn/infosupply/31407572.htm), p‐P38 (1:1,000; cat. no. bs‐2210R; BIOSS) and β‐actin (1:1,000; cat. no. bs‐0061R; BIOSS) overnight.

Techniques:

Journal: Alzheimer's & Dementia : Translational Research & Clinical Interventions

Article Title: Inhibition of Tau aggregation with BSc3094 reduces Tau and decreases cognitive deficits in rTg4510 mice

doi: 10.1002/trc2.12170

Figure Lengend Snippet: BSc3094 treatment did not reverse the loss of synaptic markers in rTg4510 mice. (A) A two‐way ANOVA showed an overall effect of the genotype on the levels of GluR1 [F(1, 22) = 35.08; P = .0001]. Further uncorrected Fisher's LSD post hoc test showed that vehicle‐treated rTg4510 mice presented significantly lower levels of GluR1 compared to vehicle‐treated control mice ( P = .0030), a decrease that was also observed in BSc3094‐treated rTg4510 mice ( P = .0039), demonstrating the BSc3094 fails to reverse the loss of GluR1 observed in the transgenic mice. (B) An overall effect of the genotype was observed on the levels of PSD95 [F(1, 22) = 8.892; P = .0066]. Uncorrected Fisher's LSD post hoc test revealed that vehicle‐treated rTg4510 mice presented significantly lower levels of PSD95 compared to vehicle‐treated control mice ( P = .0456), a decrease that was also observed in BSc3094‐treated rTg4510 mice ( P = .0153), demonstrating that the drug treatment did not reverse the loss in the expression of PSD95. (C) Analysis of the expression of synaptophysin with two‐way ANOVA showed an overall effect of the genotype [F(1, 22) = 34,24; P < .0001]. Further post hoc analysis with uncorrected Fisher's LSD test showed that vehicle‐treated rTg4510 mice presented significantly lower levels of synaptophysin compared to control mice ( P < .0001), an effect that was not reversed by BSc3094 treatment ( P < .0001). All numerical data are shown as mean ± SEM; * denotes the effect of the genotype; * P < .05; ** P < .01; **** P < .0001

Article Snippet: The following primary antibodies were used: 12E8 (1:2000, ELAN Pharmaceuticals), PHF‐1 (1:1000, kind gift from Dr. P. Davies), K9JA (1:20 000, DAKO), PSD95 (1:1000, Cell signaling), Synaptophysin (1:5000, Sigma‐Aldrich), and GluR1 (1:1000, Merck).

Techniques: Transgenic Assay, Expressing

Journal: Alzheimer's & Dementia : Translational Research & Clinical Interventions

Article Title: Inhibition of Tau aggregation with BSc3094 reduces Tau and decreases cognitive deficits in rTg4510 mice

doi: 10.1002/trc2.12170

Figure Lengend Snippet: Effect of Bsc3094 treatment on rTg4510

Article Snippet: The following primary antibodies were used: 12E8 (1:2000, ELAN Pharmaceuticals), PHF‐1 (1:1000, kind gift from Dr. P. Davies), K9JA (1:20 000, DAKO), PSD95 (1:1000, Cell signaling), Synaptophysin (1:5000, Sigma‐Aldrich), and GluR1 (1:1000, Merck).

Techniques: Activity Assay

Journal: eLife

Article Title: RNG105/caprin1, an RNA granule protein for dendritic mRNA localization, is essential for long-term memory formation

doi: 10.7554/eLife.29677

Figure Lengend Snippet: ( A and B ) Immunostaining for GluR1 ( A ) and GluR2 ( B ) in cultured neurons (9 DIV) from the cerebral cortex of E17.5 Rng105 +/+ and Rng105 −/− littermates. The neurons were cultured with (+) or without (-) TTX and APV prior to the staining. GluR1 and GluR2 staining before permeabilization (green, surface proteins), after permeabilization (magenta, intracellular and residual surface proteins), and merged images (total proteins) are shown. GluR1 and GluR2 are distributed in a punctate manner both in the soma and dendrites. The insets show magnified images of boxed areas. Arrowheads denote representative GluR1 and GluR2 puncta which were stained both before and after permeabilization (white), only before permeabilization (yellow) and only after permeabilization (blue). Scale bars, 10 µm. ( C and D ) Quantitative analysis of GluR1 and GluR2 surface expression in dendrites. C, the number of surface GluR1 puncta in dendrites normalized by the number of total GluR1 puncta (left), and fluorescence intensity of surface GluR1 puncta in dendrites normalized by GluR1 fluorescence intensity after permeabilization and in the soma (right). D, the same quantification for GluR2. Data are represented as the mean ± s.e.m. In C, n = 31 ( Rng105 +/+ , −), 35 ( Rng105 +/+ , +), 34 ( Rng105 −/− , −), and 33 ( Rng105 −/− , +) neurons from 4 experiments. In D, n = 39 ( Rng105 +/+ , −), 40 ( Rng105 +/+ , +), 39 ( Rng105 −/− , −), and 38 ( Rng105 −/− , +) neurons from 4 experiments. ***p<0.005, ****p<0.001 using two-way ANOVA followed by post-hoc Student's t-test. See also .

Article Snippet: Live neurons were incubated with an anti-GluR1 (1:15, PC246, Merck Millipore) or an anti-GluR2 antibody (1:100, MAB397, Merck Millipore) at 37°C in a 5% CO 2 incubator for 1 hr.

Techniques: Immunostaining, Cell Culture, Staining, Expressing, Fluorescence

Journal: eLife

Article Title: RNG105/caprin1, an RNA granule protein for dendritic mRNA localization, is essential for long-term memory formation

doi: 10.7554/eLife.29677

Figure Lengend Snippet: ( A ) Total cell lysates of surface-biotinylated primary cultured neurons (9 DIV) from E17.5 wild-type mouse cerebral cortex (total), and avidin agarose beads-bound fractions of the lysates (surface), were immunoblotted with the anti-GluR1 antibody. Control neurons were mock-treated without biotin. Arrow and arrowhead indicate biotin-labeled surface GluR1 and non-labeled intracellular GluR1, respectively. Lanes were cut and moved horizontally in the same membrane. ( B and C ) TTX/APV-treated and untreated primary cultured neurons (9 DIV) from Rng105 +/+ and Rng105 −/− littermates (E17.5) were surface biotinylated and analyzed as in A. Immunoblotting for GluR1 ( B ) and GluR2 ( C ). Arrows and arrowheads indicate biotin-labeled surface GluR1/2 and non-labeled intracellular GluR1/2, respectively. In the bottom panel in B, twice the amount of samples from Rng105 −/− neurons were loaded, which showed more clearly that the ratio of surface/intracellular GluR1 was lower in Rng105 −/− neurons than in Rng105 +/+ neurons. In A−C, numbers on the left indicate molecular mass (kDa). ( D and E ) Quantitative analysis of the ratio of surface/intracellular GluR1 and GluR2 in the biotinylation assay. The intensity of upper GluR1/2 bands in the avidin beads-bound fraction (arrows in B and C) and lower GluR1/2 bands in the total lysate (arrowheads in B and C) was measured and the surface/intracellular ratio was calculated. D, GluR1; E, GluR2. Data are represented as the mean ± s.e.m. n = 9 from 3 littermates each of Rng105 +/+ and Rng105 −/− mice. *p<0.05 using two-way ANOVA followed by post-hoc Student's t-test. Attached Files.

Article Snippet: Live neurons were incubated with an anti-GluR1 (1:15, PC246, Merck Millipore) or an anti-GluR2 antibody (1:100, MAB397, Merck Millipore) at 37°C in a 5% CO 2 incubator for 1 hr.

Techniques: Cell Culture, Avidin-Biotin Assay, Control, Labeling, Membrane, Western Blot, Cell Surface Biotinylation Assay