Journal: bioRxiv

Article Title: GluN2D-containing NMDA receptors regulate dentate gyrus function by facilitating granule cell activity and mediating synaptic plasticity

doi: 10.64898/2026.03.06.710109

Figure Lengend Snippet: (A) Grin2d f l/fl mice were injected with AAV5-CamKII-mCherry (Control) or AAV-CamKII-mCherry-Cre ( Grin2d cKO). NMDAR-LTP was abolished in Grin2d cKO compared with control mice (Control: 149.5 ± 6.0 %, p < 0.01, n = 5, paired t-test; cKO: 92.5 ± 5.3 %, p = 0.12201, n = 6, paired t-test; Control vs cKO: p < 0.001, unpaired t-test). (B) WT mice were bilaterally injected with an anti-GluN2D antibody or control Ab into the dentate gyrus. After one hour, animals were euthanized, and slices were prepared. Injection was confirmed by the presence of methylene blue. NMDAR-LTP was abolished in mice injected with the anti-GluN2D antibody (cKO: 110.4 ± 8.5 %, p = 0.2952, n = 6, paired t-test) compared with control mice (Control: 149.8 ± 8.1 %, p < 0.001, n = 7, paired t-test; Control vs cKO: p < 0.01, unpaired t-test). (C) NMDAR-LTP was impaired in Grid1 KO mice (KO: 117.7 ± 5.3, p < 0.05%, n = 8, Wilcoxon signed-rank test) compared with controls (Control: 147.5 ± 6.7 %, p < 0.001, n = 7, paired t-test; Control vs cKO: p < 0.05, Mann-Whitney U test). Data are presented as mean ± s.e.m.

Article Snippet: For GluN2D cross-linking experiments in C57BL/6J, the control group received 1 μL of anti-rabbit Alexa 568 (control IgG, 1/5), while the GluN2D-cross-link group received 1 μg of polyclonal rabbit anti-GluN2D subunit (Alomone Labs, cat #AGC-020), both diluted in PBS with 1% methylene blue (1 μL final volume).

Techniques: Injection, Control, MANN-WHITNEY

Journal: The Journal of physiology

Article Title: Neurones in the supraoptic nucleus of the rat are regulated by a projection from the suprachiasmatic nucleus.

doi: 10.1111/j.1469-7793.1997.149bl.x

Figure Lengend Snippet: Figure 3. PSTHs to illustrate the effects of excitatory and inhibitory antagonists on the SCN- evoked responses of three continuously firing SON cells in vitro A, the non-NMDA antagonist CNQX (10 ìÒ) reduced the excitatory effects of SCN stimulation. The NMDA antagonist APV (10 ìÒ) did not influence the effects of SCN stimulation on this cell (B), but reduced the excitatory effects of SCN stimulation on three other cells (C). D, in another cell, bicuculline (20 ìÒ) blocked the inhibitory effects of SCN stimulation. (Bin width, 5 ms for A and B, 10 ms for C and 20 ms for D; stimulus at time 0; 300 sweeps.)

Article Snippet: The following drugs were used to antagonize different neurotransmitter agents: bicuculline methiodide (Sigma) to antagonize GABAA receptors; a¬_2-amino-5-phosphonovaleric acid (APV; Sigma) to antagonize NMDA glutamate receptors; and 6-cyano-7nitroquinoxaline-2,3-dione (CNQX; Tocris Cookson) and 6-nitro-7sulphamoylbenzo(f)quinoxaline-2,3-dione (NBQX; Tocris Cookson) to antagonize non-NMDA glutamate receptors.

Techniques: In Vitro

Journal: The Journal of Neuroscience

Article Title: Functional Change of NMDA Receptors Related to Enhancement of Susceptibility to Neurotoxicity in the Developing Pontine Nucleus

doi: 10.1523/jneurosci.18-19-07941.1998

Figure Lengend Snippet: Figure 3. Effects of NMDA and non-NMDA receptor antagonists on NMDA neurotoxicity during postnatal development. Nissl-stained frontal brainstem sections are shown. A, Ketamine (0.1 mM), a noncompetitive antagonist of the NMDA receptor, was injected with 0.5 mM NMDA (total volume, 1 ml; 0.04 ml/min for 25 min) into the pontine nucleus at P14. B, NBQX (0.5 mM), a non-NMDA receptor antagonist, was injected with 0.5 mM NMDA into the pontine nucleus at P14. The animals were killed 4 d later. The lesion produced by 0.5 mM NMDA is blocked by 0.1 mM ketamine (A) but not by 0.5 mM NBQX (B); obvious neuronal loss is observed surrounding the injection site in B. Arrows indicate traces of the tip of a micropipette. Scale bar, 0.4 mm.

Article Snippet: In addition, to confirm that the lesion elicited by NMDA injection is attributable to NMDA receptor stimulation, we injected 1 mM D-2amino-5-phosphonopentanoic acid (AP-5), a competitive antagonist of the NMDA receptor (Tocris Cookson, Bristol, UK), 0.1 mM ketamine, a noncompetitive antagonist of the NMDA receptor (Sigma), or 0.5 mM 2,3-dihydroxy-6-nitro-7-sulfamoyl-benzo(F)quinoxaline (NBQX), a nonNMDA receptor antagonist (Tocris Cookson), with 0.5 mM NMDA into the pontine nucleus at P14–P16 (for each group, n 5 6).

Techniques: Staining, Injection, Produced

Journal: The Journal of Neuroscience

Article Title: Functional Change of NMDA Receptors Related to Enhancement of Susceptibility to Neurotoxicity in the Developing Pontine Nucleus

doi: 10.1523/jneurosci.18-19-07941.1998

Figure Lengend Snippet: Figure 8. Permeation of Ca 21 through NMDA receptor channels in the presence of Mg 21 during postnatal development. Neurons in the pontine nucleus at P3 (a), P14 (b), and P28 (c) were perfused with a Ca 21 Ringer’s solution in which Na 1 and K 1 were replaced with an impermeant cation NMG and 10 mM Ca 21 and 5 mM Mg 21 were added. Ionophoretic applications of NMDA were performed to evoke current responses. Current responses at 2100 mV are shown. Substantial NMDA-induced Ca 21 currents are shown at P14.

Article Snippet: In addition, to confirm that the lesion elicited by NMDA injection is attributable to NMDA receptor stimulation, we injected 1 mM D-2amino-5-phosphonopentanoic acid (AP-5), a competitive antagonist of the NMDA receptor (Tocris Cookson, Bristol, UK), 0.1 mM ketamine, a noncompetitive antagonist of the NMDA receptor (Sigma), or 0.5 mM 2,3-dihydroxy-6-nitro-7-sulfamoyl-benzo(F)quinoxaline (NBQX), a nonNMDA receptor antagonist (Tocris Cookson), with 0.5 mM NMDA into the pontine nucleus at P14–P16 (for each group, n 5 6).

Techniques:

Journal: The Journal of Neuroscience

Article Title: Functional Change of NMDA Receptors Related to Enhancement of Susceptibility to Neurotoxicity in the Developing Pontine Nucleus

doi: 10.1523/jneurosci.18-19-07941.1998

Figure Lengend Snippet: Figure 9. Changes in permeation of Ca 21 through NMDA receptor channels of developing neurons in the pontine nucleus at 2100 mV in the presence of Mg 21. The responses were recorded from neurons in the pontine nucleus at P3–P4, P14–P15, and P28–P29. The amplitude of current responses was normalized to the mean value at 150 mV in each experiment. Histograms show pooled data of peak amplitudes of current responses to NMDA in a Ca 21 Ringer’s solution. Error bars represent SEM (each column, n 5 8). The normalized peak amplitude of current responses to NMDA at P14–P15 is significantly larger than that at P3–P4 and at P28–P29 (one-way ANOVA and Bonferroni/Dunn post hoc tests, *p , 0.0001).

Article Snippet: In addition, to confirm that the lesion elicited by NMDA injection is attributable to NMDA receptor stimulation, we injected 1 mM D-2amino-5-phosphonopentanoic acid (AP-5), a competitive antagonist of the NMDA receptor (Tocris Cookson, Bristol, UK), 0.1 mM ketamine, a noncompetitive antagonist of the NMDA receptor (Sigma), or 0.5 mM 2,3-dihydroxy-6-nitro-7-sulfamoyl-benzo(F)quinoxaline (NBQX), a nonNMDA receptor antagonist (Tocris Cookson), with 0.5 mM NMDA into the pontine nucleus at P14–P16 (for each group, n 5 6).

Techniques:

Journal: The Journal of Neuroscience

Article Title: Functional Change of NMDA Receptors Related to Enhancement of Susceptibility to Neurotoxicity in the Developing Pontine Nucleus

doi: 10.1523/jneurosci.18-19-07941.1998

Figure Lengend Snippet: Figure 10. In situ hybridization showing the expression of mRNAs for the five NMDA receptor subunits (NR1, NR2A, NR2B, NR2C, and NR2D) in the developing rat pontine nucleus. A–E, NR1. F–J, NR2A. K–O, NR2B. P–T, NR2C. U–Y, NR2D. Antisense oligonucleotides labeled with 35S-dATP were hybridized to adjacent frontal sections at P1 (A, F, K, P, U), at P7 (B, G, L, Q, V), at P14 (C, H, M, R, W), at P21 (D, I, N, S, X), and in the adult (E, J, O, T, Y) in the same experiment. Arrowheads in A–E indicate the pontine nucleus. Each set of developing brainstem sections was exposed to a single x-ray film, from which photographs were directly printed at the same magnification. Scale bar, 1 mm.

Article Snippet: In addition, to confirm that the lesion elicited by NMDA injection is attributable to NMDA receptor stimulation, we injected 1 mM D-2amino-5-phosphonopentanoic acid (AP-5), a competitive antagonist of the NMDA receptor (Tocris Cookson, Bristol, UK), 0.1 mM ketamine, a noncompetitive antagonist of the NMDA receptor (Sigma), or 0.5 mM 2,3-dihydroxy-6-nitro-7-sulfamoyl-benzo(F)quinoxaline (NBQX), a nonNMDA receptor antagonist (Tocris Cookson), with 0.5 mM NMDA into the pontine nucleus at P14–P16 (for each group, n 5 6).

Techniques: In Situ Hybridization, Expressing, Labeling

Journal: Frontiers in Cell and Developmental Biology

Article Title: Astrocytic gatekeeping of neural circuitry and synaptic balance in an autism mouse model: mechanistic insights beyond Gryllus bimaculatus extract-derived therapy

doi: 10.3389/fcell.2025.1677851

Figure Lengend Snippet: Protective effects of Gryllus bimaculatus (Gb) extract on abnormal expression levels of glutamatergic and GABAergic synaptic proteins in the valproic acid (VPA)-induced autism spectrum disorder (ASD) mouse brain tissues. Immunoblot analyses for GRM5, vGluT1, NMDA R1, GABA R1α, and VGAT proteins were performed on prefrontal cortex (PFC) tissue lysates collected at embryonic day 15 (E15) (A) , postnatal day 3 (P3) (B) , and P40 (C) from mice subjected to various treatment combinations. Experimental groups included CTL (saline, n = 8); VPA (600 mg/kg VPA, n = 8); VPA + Gb 5 (600 mg/kg VPA + 5 g/kg Gb extract, n = 8); VPA + Gb 10 (600 mg/kg VPA + 10 g/kg Gb extract, n = 8); Gb 5 (5 g/kg Gb extract, n = 8); Gb 10 (10 g/kg Gb extract, n = 8). Control values were normalized to 1 (mean ± SEM, n = 3; * p < 0.05, ** p < 0.01, *** p < 0.001 compared with control; # p < 0.05, ## p < 0.01, ### p < 0.001 compared with VPA alone; ns , not significant).

Article Snippet: Antibodies against synaptic markers, including NLGN1 (#NBP2-42192), NLGN2 (#NBP2-41299), NLGN3 (#NBP2-42200), SHANK3 (#NBP1-47610), the NMDA receptor NR1 subunit (#NB300-114) from Novus Biologicals (Centennial, CO, United States) and NRXN1 (#PA5-79764) was from Thermo Fisher Scientific (Waltham, MA, United States).

Techniques: Expressing, Western Blot, Saline, Control

Journal: Frontiers in Cell and Developmental Biology

Article Title: Astrocytic gatekeeping of neural circuitry and synaptic balance in an autism mouse model: mechanistic insights beyond Gryllus bimaculatus extract-derived therapy

doi: 10.3389/fcell.2025.1677851

Figure Lengend Snippet: Regulatory effects of Gryllus bimaculatus (Gb) extract on excitatory and inhibitory neuronal activity in primary cortical neurons from valproic acid (VPA)-treated embryonic mice. (A) Schematic representation of primary cortical neuron cultures derived from embryonic mouse brains. Experimental groups included CTL (saline, n = 8); VPA (600 mg/kg VPA, n = 8); VPA + Gb 5 (600 mg/kg VPA + 5 g/kg Gb extract, n = 8); VPA + Gb 10 (600 mg/kg VPA + 10 g/kg Gb extract, n = 8); Gb 5 (5 g/kg Gb extract, n = 8); Gb 10 (10 g/kg Gb extract, n = 8). (B,D) Immunoblot analyses of NMDA R1, vGluT1, GRM5, GABA R1α, VGAT, NLGN3, NRXN1, and Tuj-1 in cultured primary cortical neuron lysates. Equal amounts of protein were loaded per lane, with β-tubulin used as a loading control. The bars represent fold-changes in the densitometric values of individual protein bands relative to the corresponding β-tubulin band densities. Control values were normalized to 1 (mean ± SEM, n = 3; * p < 0.05, ** p < 0.01, *** p < 0.001 compared with control; ## p < 0.01, ### p < 0.001 compared with VPA alone; ns , not significant). (C) Confocal microscopy images of cortical neurons from various experimental groups. Cells were cultured for 7 days, fixed, and subsequently immunostained for vGluT1 (red), with nuclei counterstained using DAPI (blue). Scale bar: 50 μm.

Article Snippet: Antibodies against synaptic markers, including NLGN1 (#NBP2-42192), NLGN2 (#NBP2-41299), NLGN3 (#NBP2-42200), SHANK3 (#NBP1-47610), the NMDA receptor NR1 subunit (#NB300-114) from Novus Biologicals (Centennial, CO, United States) and NRXN1 (#PA5-79764) was from Thermo Fisher Scientific (Waltham, MA, United States).

Techniques: Activity Assay, Derivative Assay, Saline, Western Blot, Cell Culture, Control, Confocal Microscopy

Journal: Frontiers in Cell and Developmental Biology

Article Title: Astrocytic gatekeeping of neural circuitry and synaptic balance in an autism mouse model: mechanistic insights beyond Gryllus bimaculatus extract-derived therapy

doi: 10.3389/fcell.2025.1677851

Figure Lengend Snippet: Crucial role of astrocytes in excitatory and inhibitory (E/I) neurotransporter activities in Gryllus bimaculatus (Gb) extract-treated mixed cultures from valproic acid (VPA)-treated mouse brain. (A) Schematic representation of three different types of mixed culture systems derived from embryonic and postnatal mouse brains: Type 1, astrocytes from each treatment group combined with neurons from untreated mice; Type 2, astrocytes from untreated mice combined with neurons from each treatment group; Type 3, astrocytes and neurons both derived from the same treatment group. Astrocytes from postnatal day 3 mouse brains were seeded for 7 days, followed by the addition of cortical neurons from embryonic day 15 mouse brains onto astrocytes monolayers for an additional 7 days. (B–D) Confocal microscopy images of the different types of mixed cultures. Cells were fixed and immunostained for Tuj-1 (green) and GFAP (purple), with nuclei counterstained using DAPI (blue). Scale bar: 50 μm. Experimental groups included CTL (saline, n = 8); VPA (600 mg/kg VPA, n = 8); VPA + Gb 5 (600 mg/kg VPA + 5 g/kg Gb extract, n = 8); VPA + Gb 10 (600 mg/kg VPA + 10 g/kg Gb extract, n = 8); Gb 5 (5 g/kg Gb extract, n = 8); Gb 10 (10 g/kg Gb extract, n = 8). (E) Western blots analysis of type III mixed culture. Cell lysates were immunoblotted for Tuj-1, GFAP, synaptophysin, NMDA receptor 1 (NMDA R1), GABA receptor 1α (GABA R1α), EAAT1, and EAAT2. Equal amounts of protein were loaded per each lane, with β-actin serving as the loading control. Bars represent fold-changes in the densitometric values of the bands relative to the corresponding β-actin densities. Control values were normalized to 1 (mean ± SEM, n = 3; * p < 0.05, ** p < 0.01, *** p < 0.001 compared with control; # p < 0.05, ## p < 0.01, ### p < 0.001 compared with VPA alone; ns , not significant).

Article Snippet: Antibodies against synaptic markers, including NLGN1 (#NBP2-42192), NLGN2 (#NBP2-41299), NLGN3 (#NBP2-42200), SHANK3 (#NBP1-47610), the NMDA receptor NR1 subunit (#NB300-114) from Novus Biologicals (Centennial, CO, United States) and NRXN1 (#PA5-79764) was from Thermo Fisher Scientific (Waltham, MA, United States).

Techniques: Derivative Assay, Confocal Microscopy, Saline, Western Blot, Control

Journal: Journal of Neurochemistry

Article Title: Pharmacological target sites for restoration of age‐associated deficits in NMDA receptor‐mediated norepinephrine release in brain

doi: 10.1111/jnc.16280

Figure Lengend Snippet: Characterizing the cellular localization of the NMDA receptors regulating [ 3 H] ‐NE release in young rat cortical brain slices. 1 mM Glu‐stimulated‐[ 3 H]‐NE releases in the cerebral cortex tissue slices from young rats ( n = 3–7) in the presence 1 & 3 μM of the TTX, voltage‐gated Na channel blocker, 10 μM MK‐801, and a combination of MK‐801 and TTX. Data are expressed as mean (±SEM) of net fractional release (stimulated—basal), with each data point representing a duplicate from one animal. Data were analyzed using a mixed‐effect analysis followed by Dunnett's multiple comparison test **** p < 0.0001. NE, norepinephrine; Glu, glutamate; TTX, tetrodotoxin.

Article Snippet: The NMDA receptor NR2A and NR2B subunit polyclonal antibodies were used alongside the NR1 monoclonal antibody (PhosphoSolutions, CO, USA; Cat # 1805‐NR1).

Techniques: Comparison

Journal: Journal of Neurochemistry

Article Title: Pharmacological target sites for restoration of age‐associated deficits in NMDA receptor‐mediated norepinephrine release in brain

doi: 10.1111/jnc.16280

Figure Lengend Snippet: The stimulatory effect of Glutamate Vs. NMDA on [ 3 H] ‐NE release in young rat cortical brain slices. In (a), the concentrations‐response curves of glutamate and NMDA‐stimulated [ 3 H] ‐NE releases in the cerebral cortex tissue slices from young rats ( n = 4). In (b), the 1 mM glutamate and NMDA stimulated NE release in the presence and absence of 1.2 mM magnesium in the cerebral cortex tissue slices from young rats ( n = 3). Data were analyzed using an unpaired t ‐test. * p ≤ 0.05, ** p ≤ 0.01 NE, norepinephrine; Glu, glutamate; NMDA, N‐methyl‐ d ‐aspartate; Mg 2+ , magnesium.

Article Snippet: The NMDA receptor NR2A and NR2B subunit polyclonal antibodies were used alongside the NR1 monoclonal antibody (PhosphoSolutions, CO, USA; Cat # 1805‐NR1).

Techniques:

Journal: Journal of Neurochemistry

Article Title: Pharmacological target sites for restoration of age‐associated deficits in NMDA receptor‐mediated norepinephrine release in brain

doi: 10.1111/jnc.16280

Figure Lengend Snippet: Age‐associated changes in the expression of NMDA receptors freely solubilize subunits in the cortical rat tissue homogenate. A representative Western blot for NMDA receptors subunits in the cerebral cortex in (a); and in (b) quantified results for GluN1, GluN2A, and GluN2B expressions in young and aged rats ( n = 5). Data are expressed as mean (±SEM) normalized to young rats, with each data point representing a duplicate from one animal. Data were analyzed using an unpaired t ‐test. * p ≤ 0.05. NMDA, N‐methyl‐ d ‐aspartate).

Article Snippet: The NMDA receptor NR2A and NR2B subunit polyclonal antibodies were used alongside the NR1 monoclonal antibody (PhosphoSolutions, CO, USA; Cat # 1805‐NR1).

Techniques: Expressing, Western Blot

Journal: Journal of Neurochemistry

Article Title: Pharmacological target sites for restoration of age‐associated deficits in NMDA receptor‐mediated norepinephrine release in brain

doi: 10.1111/jnc.16280

Figure Lengend Snippet: Effect of aging on [ 3 H]‐MK‐801 binding to NMDA receptors in the young (2–3 months old) and aged (18–24 months old) rat cortical tissue membrane. In (a) saturation curve of [ 3 H]‐MK‐801 binding to NMDA receptors in young rats in the presence of 10 μM Glu and Gly ( n = 4). The non‐linear least‐squares fitting of the saturation isotherm yielded K d and B max values of 1.8 nM and 970 fmol/mg of protein, respectively. Both total and non‐specific binding of [ 3 H]‐MK‐801 is shown in the curve, and the specific [ 3 H]‐MK‐801 binding is presented with 95% CI in dotted lines. Inset: Saturation data graphed as Scatchard plots. Whereas in (b), the binding of 10 nM [ 3 H]‐MK‐801 in +/− 10 μM Glu and Gly in young and aged rats ( n = 7), each performed in triplicate and repeated twice. In (c), the % increases after subtracting baseline binding from the binding in the presence of 10 μM Glu and Gly. Baseline Binding values represent [ 3 H]‐MK‐801 binding without exogenous addition of Glu and Gly. Data were analyzed using a Mixed‐effect analysis followed by Tukey's multiple comparison tests. * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001, **** p ≤ 0.0001; whereas the % of increase over the baseline was analyzed using an unpaired t ‐test: ** p ≤ 0.01. NMDA, N‐methyl‐d‐aspartate; Glu, Glutamate; Gly, Glycine; K d , dissociation constant; B max , Maximum Binding; n H , Hill Coefficient).

Article Snippet: The NMDA receptor NR2A and NR2B subunit polyclonal antibodies were used alongside the NR1 monoclonal antibody (PhosphoSolutions, CO, USA; Cat # 1805‐NR1).

Techniques: Binding Assay, Membrane, Comparison

Journal: Neural Plasticity

Article Title: Seizure-Induced Regulations of Amyloid- β , STEP 61 , and STEP 61 Substrates Involved in Hippocampal Synaptic Plasticity

doi: 10.1155/2016/2123748

Figure Lengend Snippet: A single ECS and chronic ECS differently alter Tyr 204/187 -phosphorylation of ERK1/2 in the hippocampus. Immunoblot analysis for the phosphorylation of ERK1/2 at Tyr 204/187 (Y 204/187 ) and total ERK1/2 expression in the hippocampal crude membrane (P2) fraction following a single ECS ( n = 5 rats per time point) (a) and chronic ECS ( n = 6 rats per time point) (b). The ratio of the Tyr 204/187 -phosphorylated ERK1/2 band intensity over the β -actin band intensity (top graphs) and the ratio of total ERK1/2 band intensity over the β -actin band intensity (bottom graphs) were calculated per each time point and normalized to that of “no seizure” (NS) sham group. Data shown represent the mean band intensity ± SEM. (a) A single ECS transiently decreases the level of Tyr 204/187 -phosphorylated ERK1/2 in the hippocampus at 48 h ( ### p < 0.005, t -test) and total ERK1/2 expression at 72–96 h ( ∗ p < 0.05) following a single ECS. (b) Chronic ECS significantly increases the level of Tyr 204/187 -phosphorylated ERK1/2 at 0 h following chronic ECS ( ∗ p < 0.05) but has no effect on total GluN2B expression in the hippocampus.

Article Snippet: Phosphorylation site specific antibodies used include anti-GluN2B-pTyr 1472 which recognizes phosphorylated Tyr-1472 of GluN2B (P1516-1472, PhosphoSolutions), anti-ERK1/2-pThr 202 /Tyr 204 which recognizes phosphorylated Thr 202 /Tyr 204 of ERK1 and Thr 185 /Tyr 187 of ERK2 (#9106, Cell Signaling), anti-GluA2-p3Y which recognizes phosphorylated Tyr 869 , Tyr 873 , and Tyr 876 (3Y) of GluA2 (#3921S, Cell Signaling), and anti-GluA2-pY 876 which recognizes phosphorylated Tyr 876 of GluA2 (#4027S, Cell Signaling).

Techniques: Phospho-proteomics, Western Blot, Expressing, Membrane

Journal: Neural Plasticity

Article Title: Seizure-Induced Regulations of Amyloid- β , STEP 61 , and STEP 61 Substrates Involved in Hippocampal Synaptic Plasticity

doi: 10.1155/2016/2123748

Figure Lengend Snippet: A single ECS but not chronic ECS transiently decreases the level of Tyr 1472 -phosphorylated GluN2B in the hippocampus. Immunoblot analysis for the phosphorylation of GluN2B at Tyr 1472 (Y 1472 ) and total GluN2B expression in the hippocampal crude membrane (P2) fraction following a single ECS ( n = 5 rats per time point) (a) and chronic ECS ( n = 6 rats per time point) (b). The ratio of the Tyr 1472 -phosphorylated GluN2B band intensity over the β -actin band intensity (top graphs) and the ratio of total GluN2B band intensity over the β -actin band intensity (bottom graphs) were calculated per each time point and normalized to that of “no seizure” (NS) sham group. Data shown represent the mean band intensity ± SEM. (a) A single ECS transiently decreases the level of Tyr 1472 -phosphorylated GluN2B in the hippocampus at 48 h ( ∗∗∗ p < 0.005) and 72 h ( ∗ p < 0.05) following a single ECS. (b) Chronic ECS significantly decreases total GluN2B expression over the time course of 96 h in the hippocampus ( ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.005).

Article Snippet: Phosphorylation site specific antibodies used include anti-GluN2B-pTyr 1472 which recognizes phosphorylated Tyr-1472 of GluN2B (P1516-1472, PhosphoSolutions), anti-ERK1/2-pThr 202 /Tyr 204 which recognizes phosphorylated Thr 202 /Tyr 204 of ERK1 and Thr 185 /Tyr 187 of ERK2 (#9106, Cell Signaling), anti-GluA2-p3Y which recognizes phosphorylated Tyr 869 , Tyr 873 , and Tyr 876 (3Y) of GluA2 (#3921S, Cell Signaling), and anti-GluA2-pY 876 which recognizes phosphorylated Tyr 876 of GluA2 (#4027S, Cell Signaling).

Techniques: Western Blot, Phospho-proteomics, Expressing, Membrane

Journal: Neural Plasticity

Article Title: Seizure-Induced Regulations of Amyloid- β , STEP 61 , and STEP 61 Substrates Involved in Hippocampal Synaptic Plasticity

doi: 10.1155/2016/2123748

Figure Lengend Snippet: Model by which seizure-induced changes in A β , STEP 61 , and Tyr-phosphorylation of STEP 61 lead to synaptic weakening in the hippocampus. A single ECS increases STEP 61 expression and decreases Tyr-phosphorylation of NMDAR subunit GluN2B and ERK1/2 in the hippocampus at 48 h time point, leading to synaptic weakening via NMDAR internalization and ERK1/2 inactivation. A delayed decrease in ERK1/2 expression as well as a delayed enhancement of APP and A β expression at 72–96 h following a single ECS maintains this synaptic weakening. Chronic ECS-induced increase in APP expression and A β production as well as persistent decrease in total GluN2B level leads to synaptic weakening.

Article Snippet: Phosphorylation site specific antibodies used include anti-GluN2B-pTyr 1472 which recognizes phosphorylated Tyr-1472 of GluN2B (P1516-1472, PhosphoSolutions), anti-ERK1/2-pThr 202 /Tyr 204 which recognizes phosphorylated Thr 202 /Tyr 204 of ERK1 and Thr 185 /Tyr 187 of ERK2 (#9106, Cell Signaling), anti-GluA2-p3Y which recognizes phosphorylated Tyr 869 , Tyr 873 , and Tyr 876 (3Y) of GluA2 (#3921S, Cell Signaling), and anti-GluA2-pY 876 which recognizes phosphorylated Tyr 876 of GluA2 (#4027S, Cell Signaling).

Techniques: Phospho-proteomics, Expressing