anti glun2c  (Alomone Labs)

Journal: Molecular Brain

Article Title: NMDA GluN2C/2D receptors contribute to synaptic regulation and plasticity in the anterior cingulate cortex of adult mice

doi: 10.1186/s13041-021-00744-3

Figure Lengend Snippet: The GluN2C/2D components of the NMDAR-mediated EPSCs are significant in neurons of the ACC by using UBP145 as the antagonist of GluN2C/2D. a A selective GluN2C/2D antagonist, UBP145, partially inhibited NMDAR-mediated EPSCs. Left: the time course of changes in EPSC amplitude before, during and after the application of UBP145 (3 μM) in ACC neurons from adult male mice is shown. Traces show the currents at different time points during application of drugs. UBP145 produced its maximal effect at 20 min after bath application and was partially washed out (n = 5 neurons/3 mice). Right: Summary of the partial inhibition of the UBP145 on the NMDAR-mediated current. Data within the last five minutes of the baseline, UBP145 application and washout phase is averaged. Open circles represent the individual data points. b Summarized data of the effect of UBP145 to the rising time (left) and decay time (right) of the NMDAR-mediated EPSCs in the ACC neurons. The application of UBP145 produced a significant inhibitory effect on the decay time of EPSCs. c Left: application of the antagonist of the GluN2A (PEAQX) and GluN2B (Ro 25-6981) reduced over 60% of the NMDAR current. UBP145 further decreased the amplitude of EPSC. The remaining current were gradually decreased by the application of AP-5. Right: Summary of the gradual inhibition of the GluN2A/2B and GluN2C/2D on the NMDAR-mediated current. * p < 0.05, *** p < 0.001, compared with baseline; # p < 0.05, compared with PEAQX + Ro25-6981. Error bars represent SEM

Article Snippet: Blots were probed with anti-GluN2A (1:1000; Millipore, US), anti-GluN2B (1:500; Millipore, US), anti-GluN2C (1:500; Millipore, US) or anti-GluN2D (1:500; Millipore, US) polyclonal antibodies overnight at 4 °C.

Techniques: Produced, Inhibition

Journal: Molecular Brain

Article Title: NMDA GluN2C/2D receptors contribute to synaptic regulation and plasticity in the anterior cingulate cortex of adult mice

doi: 10.1186/s13041-021-00744-3

Figure Lengend Snippet: The GluN2C/2D components of the NMDAR-mediated EPSCs are significant in neurons of the CA1 by using UBP145 as antagonist of GluN2C/2D. a UBP145 partially inhibited NMDAR-mediated EPSCs in the CA1. Left: traces show the currents at different time points during application of drugs. UBP145 produced its maximal effect at 20 min after bath application (n = 4 neurons/3 mice). Right: Summary of the partial inhibition of the UBP145 on the NMDAR-mediated current. Data within the last five minutes of the baseline, UBP145 application and washout phase is averaged. Open circles represent the individual data points. b Summarized data of the effect of UBP145 to the rising time (left) and decay time (right) of the NMDAR-mediated EPSCs in the CA1 neurons. The application of UBP145 produced a significant inhibitory effect on the decay time of EPSCs. ** p < 0.01. *** p < 0.001. Error bars represent SEM

Article Snippet: Blots were probed with anti-GluN2A (1:1000; Millipore, US), anti-GluN2B (1:500; Millipore, US), anti-GluN2C (1:500; Millipore, US) or anti-GluN2D (1:500; Millipore, US) polyclonal antibodies overnight at 4 °C.

Techniques: Produced, Inhibition

Journal: Molecular Brain

Article Title: NMDA GluN2C/2D receptors contribute to synaptic regulation and plasticity in the anterior cingulate cortex of adult mice

doi: 10.1186/s13041-021-00744-3

Figure Lengend Snippet: Effects of GluN2C/2D antagonists to the synaptic plasticity in the ACC. a Left: LTP was induced in pyramidal neurons in adult ACC (black squares, n = 5 neurons/ 3 mice) by the pairing training protocol (indicated by an arrow). Pretreated slices with 3 μM UBP145 didn’t affect the induction of LTP (red circles, n = 4 neurons/ 3 mice). Sample traces of evoked EPSCs during baseline (1) and 30 min after the induction stimulus (2) are showed on the top; Right: Summary of the effect of the UBP145 on the postsynaptic LTP. Data within the last five minutes of the baseline and 30 min after the induction are averaged. b Top: pre-LTP was induced in pyramidal neurons in the ACC of adult mice (black squares, n = 4 neurons/ 3 mice) by giving 240 pulses at 2 Hz (indicated by an arrow). Pretreated slices with 3 μM UBP145 did not affect the induction of pre-LTP (red circles, n = 4 neurons/ 3 mice). Sample traces of evoked EPSCs with paired-pulse stimulation at 50 ms at a holding membrane potential of -70 mV during baseline (1) and 30 min after the induction stimulus (2) are showed on the top; Bottom: PPR values for the control (black squares) and UBP145 group (red circles). c LTD was induced in pyramidal neurons in adult ACC (black squares, n = 5 neurons/ 3 mice) by the pairing training protocol (300 pulses at 1 Hz while holding at -45 mV indicated by an arrow). Pretreated slices with 3 μM UBP145 did not affect the induction of LTD (red circles, n = 4 neurons/ 3 mice). Sample traces of evoked EPSCs during baseline (1) and 30 min after the induction stimulus (2) are showed on the top. ** p < 0.01, *** p < 0.001. Error bars represent SEM

Article Snippet: Blots were probed with anti-GluN2A (1:1000; Millipore, US), anti-GluN2B (1:500; Millipore, US), anti-GluN2C (1:500; Millipore, US) or anti-GluN2D (1:500; Millipore, US) polyclonal antibodies overnight at 4 °C.

Techniques:

Journal: Molecular Brain

Article Title: NMDA GluN2C/2D receptors contribute to synaptic regulation and plasticity in the anterior cingulate cortex of adult mice

doi: 10.1186/s13041-021-00744-3

Figure Lengend Snippet: GluN2C/2D modulate spontaneous presynaptic release in the ACC. a Top: Representative traces of the mEPSCs recorded in the ACC neurons before and after applying PEAQX (0.4 μM), a selective antagonist of GluN2A. Bottom: Statistic results of the frequency (left) and amplitude (right) of mEPSCs (n = 5 neurons/3 mice). b Top: Representative traces of the mEPSCs recorded in the ACC neurons before and after applied Ro 25-6981 (3 μM), a selective antagonist of GluN2B. Bottom: Statistic results of the frequency (left) and amplitude (right) of mEPSCs (n = 7 neurons/6 mice). c Top: Representative traces of the mEPSCs recorded in the ACC neurons before and after applying UBP145 (3 μM). Bottom: Statistic results of the frequency (left) and amplitude (right) of mEPSCs (n = 10 neurons/4 mice). d Top: Representative traces of the mEPSCs recorded in the ACC neurons before and after applied PPDA (10 μM). Bottom: Statistic results of the frequency (left) and amplitude (right) of mEPSCs (n = 5 neurons/3 mice). Open circles represent the individual data points. * p < 0.05, error bars indicated SEM

Article Snippet: Blots were probed with anti-GluN2A (1:1000; Millipore, US), anti-GluN2B (1:500; Millipore, US), anti-GluN2C (1:500; Millipore, US) or anti-GluN2D (1:500; Millipore, US) polyclonal antibodies overnight at 4 °C.

Techniques:

Journal: Translational Psychiatry

Article Title: Environmental enrichment prevents the late effect of acute stress-induced fear extinction deficit: the role of hippocampal AMPA-GluA1 phosphorylation

doi: 10.1038/s41398-020-01140-6

Figure Lengend Snippet: Stressed SC-housed animals showed higher expression of pGluA1 ( A ) and GluA1 ( B ) in the HC comparing to SC-housed, unstressed (only in B ), EE-housed, stressed and unstressed animals. EE-housed animals (both stressed and unstressed) presented the same expression levels of pGluA1 ( A ) and GluA1 ( B ) than SC-housed, unstressed animals. There were no differences in the GluN2B ( C ) and GluN2C ( D ) expression in both SC- and EE-housed animals (stressed or unstressed). There were no differences in the synaptophysin ( E ) and PSD-95 ( F ) expression in both SC- and EE-housed animals, regardless of the presence of stress. Representative autoradiography of western blot ( G ). Results are represented as mean ± SEM ( n = 5–6 animals per group in A ; n = 6 for each group in B , D – F ; n = 3 for each group in C ). Two-way ANOVA followed by Tukey’s multiple comparisons test. Significance differences between groups are indicated as * p < 0.05 and ** p < 0.01.

Article Snippet: Blots were blocked with 5% bovine serum albumin (BSA) diluted in TBS-T buffer (50 mM Tris-HCl, 150 mM NaCl, 0.1% Tween 20, pH 7.5) for 1 h at room temperature, and subsequently incubated overnight at 4 °C with specific primary antibodies: polyclonal rabbit anti-GluA1 (AB1504, dilution 1:2000; EMD Millipore Corporation), polyclonal rabbit anti-Ser845phospho-GluA1 (AB5849, dilution 1:1000; EMD Millipore Corporation), polyclonal rabbit anti-GluN2B (#454582, dilution 1:1000; EMD Millipore Corporation), polyclonal rabbit anti-GluN2C (#454584, dilution 1:1000; EMD Millipore Corporation), monoclonal mouse anti-synaptophysin (AB8049, dilution of 1:20,000; Sigma-Aldrich Corporation), and monoclonal mouse anti-PSD-95 (AB99099, dilution of 1:20,000; Abcam).

Techniques: Expressing, Autoradiography, Western Blot

Journal: Translational Psychiatry

Article Title: Environmental enrichment prevents the late effect of acute stress-induced fear extinction deficit: the role of hippocampal AMPA-GluA1 phosphorylation

doi: 10.1038/s41398-020-01140-6

Figure Lengend Snippet: There was an environmental effect on the pGluA1 ( A ) expression in the BLA, but there were no effects of either environment or stress on the expression of GluA1( B ), GluN2B ( C ), GluN2C ( D ), synaptophysin ( E ), and PSD-95 ( F ) in the same brain area. Representative autoradiography of western blot ( G ). Results are represented as mean ± SEM ( n = 5–6 animals per group in A and B ; n = 6 for each group in D – F ; n = 3 for each group in C ). Two-way ANOVA followed by Tukey’s multiple comparisons test (no differences were detected between the experimental groups). ANOVA significant main effect is indicated as ## p < 0.01.

Article Snippet: Blots were blocked with 5% bovine serum albumin (BSA) diluted in TBS-T buffer (50 mM Tris-HCl, 150 mM NaCl, 0.1% Tween 20, pH 7.5) for 1 h at room temperature, and subsequently incubated overnight at 4 °C with specific primary antibodies: polyclonal rabbit anti-GluA1 (AB1504, dilution 1:2000; EMD Millipore Corporation), polyclonal rabbit anti-Ser845phospho-GluA1 (AB5849, dilution 1:1000; EMD Millipore Corporation), polyclonal rabbit anti-GluN2B (#454582, dilution 1:1000; EMD Millipore Corporation), polyclonal rabbit anti-GluN2C (#454584, dilution 1:1000; EMD Millipore Corporation), monoclonal mouse anti-synaptophysin (AB8049, dilution of 1:20,000; Sigma-Aldrich Corporation), and monoclonal mouse anti-PSD-95 (AB99099, dilution of 1:20,000; Abcam).

Techniques: Expressing, Autoradiography, Western Blot

Journal: Translational Psychiatry

Article Title: Environmental enrichment prevents the late effect of acute stress-induced fear extinction deficit: the role of hippocampal AMPA-GluA1 phosphorylation

doi: 10.1038/s41398-020-01140-6

Figure Lengend Snippet: There was an environmental effect on the pGluA1 ( A ) expression in the FC, but there were no effects of either environment or stress on the expression of GluA1 ( B ), GluN2B ( C ), GluN2C ( D ), synaptophysin ( E ), and PSD-95 ( F ) in the same brain area. Representative autoradiography of western blot ( G ). Results are represented as mean ± SEM ( n = 5–6 animals per group in A and B ; n = 6 for each group in D – F ; n = 3 for each group in C ). Two-way ANOVA followed by Tukey’s multiple comparisons test (no differences were detected between the experimental groups). ANOVA significant main effect is indicated as # p < 0.05.

Article Snippet: Blots were blocked with 5% bovine serum albumin (BSA) diluted in TBS-T buffer (50 mM Tris-HCl, 150 mM NaCl, 0.1% Tween 20, pH 7.5) for 1 h at room temperature, and subsequently incubated overnight at 4 °C with specific primary antibodies: polyclonal rabbit anti-GluA1 (AB1504, dilution 1:2000; EMD Millipore Corporation), polyclonal rabbit anti-Ser845phospho-GluA1 (AB5849, dilution 1:1000; EMD Millipore Corporation), polyclonal rabbit anti-GluN2B (#454582, dilution 1:1000; EMD Millipore Corporation), polyclonal rabbit anti-GluN2C (#454584, dilution 1:1000; EMD Millipore Corporation), monoclonal mouse anti-synaptophysin (AB8049, dilution of 1:20,000; Sigma-Aldrich Corporation), and monoclonal mouse anti-PSD-95 (AB99099, dilution of 1:20,000; Abcam).

Techniques: Expressing, Autoradiography, Western Blot