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: 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: International Journal of Molecular Sciences

Article Title: AMPA and NMDA Receptors in Hippocampus of Rats with Fluoride-Induced Cognitive Decline

doi: 10.3390/ijms252111796

Figure Lengend Snippet: mRNA and protein levels of GluN1 subunits of NMDARs in hippocampal cells. ( A ) Relative expression of Grin1 gene in the hippocampus of rats consumed different F - doses normalized to that of a pair of reference genes ( Eef1a+Ppia ). Average values ± SEM (n = 10). ( B , C ) Protein expression of native and phosphorylated (Ser890) forms of GluN1 subunit in cytosolic and membrane fractions, respectively. Shown are typical immunoblots and average values ± SEM (n = 8).

Article Snippet: Rabbit antibodies to GluA1 (#13185), phospho-GluA1 (#8084), GluA2 (#5306), phospho-GluA2 (#3921), GluA3 (#4676), GluN1 (#5704), phospho-GluN1 (#3381), GluN2A (#4205), phospho-GluN2A (#4206), GluN2B (#4207) and phospho-GluN2B (#4208) were purchased from Cell Signaling Technology (Danvers, MA, USA).

Techniques: Expressing, Membrane, Western Blot

Journal: bioRxiv

Article Title: A Consequence of Immature Breathing induces Persistent Changes in Hippocampal Synaptic Plasticity and Behavior: A Role of Pro-Oxidant State and NMDA Receptor Imbalance

doi: 10.1101/2023.03.21.533692

Figure Lengend Snippet: A . ( left ) Representative traces of the evoked fEPSP from control (black), control+AP5 [50µM] (green), nIH (red) and nIH +AP5 [50µM] (blue) in baseline conditions prior to TBS (1) and following TBS (2). ( middle ) Mean fEPSP slope plotted as a function of time relative to the slope before TBS in control, control+AP5, nIH and nIH+AP5. ( right ) fEPSP slope represented as percent change from baseline at 60 min after TBS in control slices vs control+AP5 [50 µM]. B . ( top ) Representative image for GluN1. (bottom) The graph shows GluN1 did not change protein expression after nIH exposure compared with control. (two tailed t-test, t=0.63; df=4.1; P=0.55). C . (top) Western blot picture for GluN2A. (bottom) Comparison for both conditions show GluN2A decreased the protein content levels after nIH. (two tailed t-test, t=4.017; df=6.43; P=0.006). D . ( top ) Representative immunoblot image for GluN2B. (bottom) Comparison of both conditions show increased GluN2B levels after nIH (two tailed t-test, t=3.43; df=5.78; P=0.014). E . GluN2B/GluN2A comparison ratio. F . ( left ) Representative traces of the evoked fEPSP control +TCN [5 µM] (orange), control + ifenprodil [5 µM] (cyan) and control + both drugs (olive) in baseline conditions prior to TBS (1) and following TBS (2). ( middle ) Mean fEPSP slope plotted as a function of time relative to the slope before TBS and ( right ) Comparison of fEPSP slope represented as percent change from baseline at 60 min after TBS (one way ANOVA, F (3,20) =43.52; P<0.001). Black dashed line represents the mean slope of the fEPSP 60 min following TBS in control slices from . G . ( left ) Representative traces of the evoked fEPSP from nIH+TCN [5 µM] (dark yellow) and nIH +ifenprodil [5 µM] (light blue) in baseline conditions prior to TBS (1) and following TBS (2). ( middle ) Mean fEPSP slope plotted as a function of time and relative to slope before TBS ( right ) fEPSP slope represented as percent change from baseline at 60 min after TBS (two tailed t-test, t=12.46; df=4.59; P=0.001). Red dashed line represents the mean slope of the fEPSP 60 min following TBS in nIH slices from . Scale bars for A, F and G: 10 msec x 0.2 mV. The box-plot parameters indicate mean ± S.E. The analysis was performed for A to E using unpaired two-tailed t-test with Welch’s correction and for F and G the analysis was performed using one-way ANOVA followed by Bonferroni post hoc. **P<0.01, ***P<0.001 and ****P<0.0001).

Article Snippet: Membranes were incubated under constant shaking with primary antibodies: anti rabbit GluN1 (1:2000; Abcam Cat# ab109182, RRID:AB_10862307) anti-rabbit GluN2A (1:2000; Cell Signaling Technology Cat# 4205, RRID:AB_2112295), anti-rabbit GluN2B (1:2000; Cell Signaling Technology Cat# 14544, RRID:AB_2798506), anti-rabbit NOX2 (1:2000; Abcam Cat# ab129068, RRID:AB_11144496) anti-rabbit NOX4 (1:500; Novus Cat# NB110-58851B, RRID:AB_1217375 and anti-mouse GAPDH (1: 10.000; Abcam Cat# ab8245, RRID:AB_2107448).

Techniques: Control, Expressing, Two Tailed Test, Western Blot, Comparison

Journal: bioRxiv

Article Title: A Consequence of Immature Breathing induces Persistent Changes in Hippocampal Synaptic Plasticity and Behavior: A Role of Pro-Oxidant State and NMDA Receptor Imbalance

doi: 10.1101/2023.03.21.533692

Figure Lengend Snippet: A . Malondialdehyde (MDA) content was measured in hippocampal homogenates from control, nIH Saline and 10-Mn. (one way ANOVA, F (2,12) =11.53; P=0.0016). B . (top) Representative blot of nuclear HIF1a performed from control, nIH Saline and IH 10-Mn mice. (bottom) Quantification of HIF1a expression (one way ANOVA, F (2,12) =5.76; P=0.017). C . (top) Immunoblot of NOX2. (bottom) Significant differences was found in hippocampal homogenate from nIH Saline compared to control and nIH Mn (one way ANOVA, F (2,15) =9.26; P=0.0024). D . (top) Representative image of NOX4. ( bottom ) Comparison of NOX4 expression between control, nIH Saline and nIH Mn . (one way ANOVA, F (2,9) =11.46; P=0.0034). E . ( left ) Representative blot of GluN2A from control, nIH Saline and nIH Mn mice. ( right ). Quantification of GluN2A expression. (one way ANOVA, F (2,15) =6.63; P=0.0086). F . ( left ) Immunoblot of GluN2B. ( right ) Significant differences were found in hippocampal homogenate from nIH Saline vs control and nIH Mn . (one way ANOVA, F (2,12) =6.88; P=0.01). G . (top) Representative traces of evoked fEPSP from nIH Mn (purple) in baseline conditions prior to TBS (1) and following TBS (2). ( left bottom ) Mean fEPSP slope plotted as a function of time relative to the slope before TBS in nIH Mn . ( right ) fEPSP slope represented as percent change from baseline at 60 min after TBS in nIH vs nIH Mn slices. (two tailed t-test, t=3.41; df=10.61; P=0.0061). Dashed lines represent the mean slope of the fEPSP 60 min following TBS in control (black dashed line) and nIH (red dashed line) slices from . H . (top) Representative traces of the evoked fEPSP from nIH Mn in presence the ifenprodil [5 µM] (light purple) in baseline conditions prior to TBS (1) and following TBS (2). ( left bottom ) Mean fEPSP slope plotted as a function of time relative to the slope before TBS in nIH Mn in presence the ifenprodil. ( right ) fEPSP slope represented as percent change from baseline at 60 min after TBS in IH+ifenprodil vs nIH Mn in presence of ifenprodil. (two tailed t-test, t=3.99; df=6.99; P=0.019). Blue dashed line represents the mean slope of the fEPSP 60 min following TBS in ifenprodil treated nIH slices from . For G and H, Scale bars 10 msec x 0.2 mV. The analysis was performed for A to F using one-way ANOVA followed by Bonferroni post hoc and for G and F, the analysis was performed using unpaired two-tailed t-test with Welch’s correction. *P<0.05, **P<0.01, and ***P<0.001.

Article Snippet: Membranes were incubated under constant shaking with primary antibodies: anti rabbit GluN1 (1:2000; Abcam Cat# ab109182, RRID:AB_10862307) anti-rabbit GluN2A (1:2000; Cell Signaling Technology Cat# 4205, RRID:AB_2112295), anti-rabbit GluN2B (1:2000; Cell Signaling Technology Cat# 14544, RRID:AB_2798506), anti-rabbit NOX2 (1:2000; Abcam Cat# ab129068, RRID:AB_11144496) anti-rabbit NOX4 (1:500; Novus Cat# NB110-58851B, RRID:AB_1217375 and anti-mouse GAPDH (1: 10.000; Abcam Cat# ab8245, RRID:AB_2107448).

Techniques: Control, Saline, Expressing, Western Blot, Comparison, Two Tailed Test

Journal: bioRxiv

Article Title: A Consequence of Immature Breathing induces Persistent Changes in Hippocampal Synaptic Plasticity and Behavior: A Role of Pro-Oxidant State and NMDA Receptor Imbalance

doi: 10.1101/2023.03.21.533692

Figure Lengend Snippet: A . (top) Representative traces of the evoked fEPSP from adult control (black) and adult mice were exposed to neonatal IH (red) in baseline conditions prior to TBS (1) and following TBS (2). ( left bottom ) Mean fEPSP slope plotted as a function of time and relative to baseline prior to TBS. ( right bottom ) fEPSP slope represented as percent change from baseline at 60 min following TBS in adult control vs Adult nIH (two tailed t-test, t=5.70; df=9.04; P=0.003). B . (top) Representative trace of the evoked from control+ TCN-213 [5 µM] and control + ifenprodil [5 µM] in baseline conditions prior to TBS (1) and following TBS (2). ( left bottom ) Mean fEPSP slope plotted as a function of time and relative to baseline prior to TBS. ( right bottom ) fEPSP comparison at 60 min following TBS in control +TCN and control+ ifenprodil (two tailed t-test, t=7.43; df=4.65; P=0.0009). Black dashed line represents the mean slope of the fEPSP 60 min following TBS in control slices from . C . (top) Representative trace of the evoked response from Adult nIH + TCN-213 [5 µM] and Adult nIH + ifenprodil [5 µM] in baseline conditions prior to TBS (1) and following TBS (2). ( left bottom ) Mean fEPSP slope plotted as a function of time relative to baseline prior to TBS. ( right bottom ) fEPSP slope represented as percent change from baseline at 60 min following TBS (two tailed t-test, t=4.72; df=4.06; P=0.0064). Red dashed line represents the mean slope of the fEPSP after 60 min following TBS in control slices from . D . (top) Representative traces of the evoked fEPSP from adult mice was exposure to neonatal IH+10 days of MnTMPyP (Adult nIH-Mn ) and adult mice receive MnTMPyP after exposure to neonatal IH (Adult REC-Mn ) in baseline conditions before TBS (1) and after TBS (2). ( left bottom ) Mean fEPSP slope plotted as a function of time and relative to slope before TBS in Adult nIH-Mn vs Adult REC-Mn ( right bottom ) fEPSP slope represented as percent change from baseline at 60 min after TBS Adult nIH-Mn vs Adult REC-Mn (two tailed t-test, t=3.79; df=8.16; P=0.0051). E. Representative image of GluN1. ( bottom ) Quantification shows GluN1 protein expression is not changed in Adult nIH , Adult nIH-Mn or 10+Mn exposures compared to control (one way ANOVA, F (3,16)=1.14 ; P=0.93; N=5). F . (top) Representative blot of GluN2A performed from adult mice unexposed, Adult nIH , Adult nIH-Mn or Adult REC-Mn . ( bottom ). Quantification of GluN2A expression from adult control, Adult nIH , Adult nIH-Mn or 10+Mn. (one way ANOVA, F (3,12)=8.31 ; P=0.0029, N=4). G . (top) Immunoblot of GluN2B. ( bottom ) Significant differences were found in hippocampal homogenates from adult control, Adult nIH , Adult nIH-Mn or Adult REC-Mn (one way ANOVA, F (3,12)=4.91 ; P=0.018, N=4). The box plot parameters indicate mean ± S.E. The analysis was performed for A-D using unpaired two-tailed t-test with Welch’s correction. The analysis was performed for E-G using one-way ANOVA followed by Bonferroni post hoc. *P=0.05, **P=0.01, ***P=0.001 and N.S= no significant. Scale bars for A,B, F and G= 10 msec x 0.2 mV

Article Snippet: Membranes were incubated under constant shaking with primary antibodies: anti rabbit GluN1 (1:2000; Abcam Cat# ab109182, RRID:AB_10862307) anti-rabbit GluN2A (1:2000; Cell Signaling Technology Cat# 4205, RRID:AB_2112295), anti-rabbit GluN2B (1:2000; Cell Signaling Technology Cat# 14544, RRID:AB_2798506), anti-rabbit NOX2 (1:2000; Abcam Cat# ab129068, RRID:AB_11144496) anti-rabbit NOX4 (1:500; Novus Cat# NB110-58851B, RRID:AB_1217375 and anti-mouse GAPDH (1: 10.000; Abcam Cat# ab8245, RRID:AB_2107448).

Techniques: Control, Two Tailed Test, Comparison, Expressing, Western Blot

Journal: Neurobiology of disease

Article Title: Ethanol exposure alters Alzheimer’s-related pathology, behavior, and metabolism in APP/PS1 mice

doi: 10.1016/j.nbd.2022.105967

Figure Lengend Snippet: Chronic moderate drinking differentially alters NMDA and GABAA receptors in the cortex and hippocampus of APP/PS1 mice. a) Ethanol treatment did not alter cortical Grin2a expression in wildtype or APP/PS1 mice. b) Ethanol-treated APP/PS1 mice had higher cortical Grin2b expression compared to EtOH-treated wildtype mice. 2-way ANOVA revealed a significant treatment × genotype interaction ( p = 0.0319). c) H 2 O-treated APP/PS1 mice showed increased cortical Gabra5 expression compared to H 2 O-exposed wildtype mice (p < 0.05). This effect was lost in EtOH-exposed APP/PS1 Gabra5 mRNA levels. 2-way ANOVA revealed a significant treatment × genotype interaction ( p = 0.0249) and a trend in genotype effects ( p = 0.0723). d) Synaptic GluN2A levels was unaltered in the hippocampus of H 2 O- or EtOH-treated wildtype or APP/PS1 mice. e) Synaptic GluN2B levels was unaltered in the hippocampus of H 2 O- or EtOH-treated wildtype or APP/PS1 mice. f) Ethanol-treated wildtype mice showed increased synaptic GABA A R α5 subunit levels compared to H 2 O-treated wildtype mice. Ethanol treatment had no effect on GABAAR α5 subunit levels in APP/PS1 mice. 2-way ANOVA revealed a significant treatment × genotype effect ( p = 0.0347) and a trend in treatment effects ( p = 0.0644). Wildtype + H2O, n = 10; APP/PS1 + H2O, n = 9; Wildtype + EtOH, n = 7; APP/PS1 + EtOH, n = 8. * p < 0.05.

Article Snippet: The following primary and secondary antibodies were used for this study: APP (including CTFβ and CTFα; Invitrogen; CT695; 1:1000), BACE1 (Cell Signaling; 5606S; 1:1000), ADAM10 (Millipore; AB19026; 1:1000), IDE (Abcam; ab232216; 1:1000), GluN2A (Cell Signaling; 4025; 1:1000), GluN2B (Cell Signaling; 4212; 1:1000), GABAAR α5 (Santa Cruz; Sc393921; 1:1000), and β-actin (Millipore; MAB1501; 1:50,000), anti-mouse (Cell Signaling; 7076S; 1:5000), anti-rabbit (Cell Signaling; 7074S; 1:5000).

Techniques: Expressing

Journal: Journal of Affective Disorders Reports

Article Title: Long-term cyclosporine A treatment promotes anxiety-like behavior: Possible relation with glutamate signaling in rat hippocampus

doi: 10.1016/j.jadr.2022.100394

Figure Lengend Snippet: Fig. 4. Western blot analysis of NMDA receptors in the hippocampus of control and CsA-treated rats. (A) Representative images of the levels of total (pan) GluN2, GluN2A, GluN2B, p-GluN1 (Ser897) and total GluN1. (B-F) Respective quantifications of immunodetections normalized to β-actin and presented as a percentage of the control mean. Data represent the mean ± SEM. Statistics: ** *p ≤0.001 as determined by Student’s t-test; n = 6.

Article Snippet: The utilized primary antibodies and respective dilutions were as follows: anti-AMPAR (1:500, #13185 Cell Signaling Technology), phosphorylated AMPAR (pAMPAR-S831) (1:1000, A4352-Sigma–Aldrich), pAMPAR (S845) (1:1000, #8084 – Cell Signaling Technology), GluN1 (1:750, #G8913 – Sigma–Aldrich), pan-GluN2 (1:750, 244-0P – SYSY Synaptic Systems), GluN2A (1:1000, #4205 - Cell Signaling Technology), GluN2B (1:1000, #4207 - Cell Signaling Technology), pGluN1 (Ser897) (1:500, 3385S – Cell Signaling Technology), nNOS (1:500, SC648 – Santa Cruz Biotechnology), and iNOS (1:750, SC7271 – Santa Cruz Biotechnology). β-actin antibody (1:5000, A5441 – Sigma-Aldrich) was used as a loading control in the experiments.

Techniques: Western Blot, Control