Journal:

Article Title: PSD-93 Deficiency Protects Cultured Cortical Neurons from NMDA Receptor-triggered Neurotoxicity

doi: 10.1016/j.neuroscience.2010.01.030

Figure Lengend Snippet: PSD-93 deficiency or knockdown protects against NMDA-stimulated neurotoxicity. Cortical neurons were cultured from WT and PSD-93 KO mice and treated with NMDA (0, 10, 20, 30, 40, and 60 μM) in the presence of 10 μM CNQX and 2 μM nimodipine. (A) Neuronal viability was assessed by MTT assay. (B) Percentage of neuronal death was determined by propidium iodide and calcein AM staining. * P < 0.05, ** P < 0.01 vs corresponding WT. n = 6 repeats. (C) Representative photographs showing propidium iodide and calcein AM staining of cortical neurons cultured from WT and PSD-93 KO mice treated with or without 30 μM NMDA or 10 μM MK-801+ 30 μM NMDA. Scale bar: 60 μm. (D) acute knock down PSD-93 by siP3 can inhibit the toxicity induced by NMDA at the dose of 30μM. MTT assay shows the cell viability reduced by 44.12% **P<0.01 vs normal group. However, siP3 can alleviate this toxicity, the cell viability increase 17%,# P<0.05 vs group transfected by siLuc.

Article Snippet: To examine whether NMDA-induced neurotoxicity was dependent on the NMDA receptor/NO signaling pathway, the cultured neurons were pre-incubated for 60 min with 10 μM MK-801 (an NMDA antagonist; Tocris, Ellisville, MI) or 30-60 μM N -Nitro-L-arginine methyl ester hydrochloride (L-NAME, a NOS inhibitor; Sigma, St. Louis, MO) at 37°C.

Techniques: Knockdown, Cell Culture, MTT Assay, Staining, Transfection

Journal:

Article Title: PSD-93 Deficiency Protects Cultured Cortical Neurons from NMDA Receptor-triggered Neurotoxicity

doi: 10.1016/j.neuroscience.2010.01.030

Figure Lengend Snippet: PSD-93 deficiency has no effect on non-NMDA receptor-triggered neurotoxicity in cultured cortical neurons. Cultured neurons were treated with kainate at the doses shown in the presence of 10 μM MK-801 and 2 μM nimodipine. Percentage of neuronal death was determined by propidium iodide and calcein AM staining (top) and neuronal viability by MTT assay (bottom). n = 6 repeats.

Article Snippet: To examine whether NMDA-induced neurotoxicity was dependent on the NMDA receptor/NO signaling pathway, the cultured neurons were pre-incubated for 60 min with 10 μM MK-801 (an NMDA antagonist; Tocris, Ellisville, MI) or 30-60 μM N -Nitro-L-arginine methyl ester hydrochloride (L-NAME, a NOS inhibitor; Sigma, St. Louis, MO) at 37°C.

Techniques: Cell Culture, Staining, MTT Assay

Journal:

Article Title: PSD-93 Deficiency Protects Cultured Cortical Neurons from NMDA Receptor-triggered Neurotoxicity

doi: 10.1016/j.neuroscience.2010.01.030

Figure Lengend Snippet: MK-801 attenuates NMDA-induced neurotoxicity in cultured cortical neurons from WT but not from PSD-93 KO mice. Percentage of neuronal death was determined by propidium iodide and calcein AM staining (top) and neuronal viability by MTT assay (bottom). * P < 0.05 vs the corresponding NMDA concentration alone. n = 6 repeats.

Article Snippet: To examine whether NMDA-induced neurotoxicity was dependent on the NMDA receptor/NO signaling pathway, the cultured neurons were pre-incubated for 60 min with 10 μM MK-801 (an NMDA antagonist; Tocris, Ellisville, MI) or 30-60 μM N -Nitro-L-arginine methyl ester hydrochloride (L-NAME, a NOS inhibitor; Sigma, St. Louis, MO) at 37°C.

Techniques: Cell Culture, Staining, MTT Assay, Concentration Assay

Journal:

Article Title: PSD-93 Deficiency Protects Cultured Cortical Neurons from NMDA Receptor-triggered Neurotoxicity

doi: 10.1016/j.neuroscience.2010.01.030

Figure Lengend Snippet: PSD-93 deficiency attenuates NMDA-stimulated Ca2+ loading in cultured cortical neurons. Cultures were challenged for 5 or 10 min with 30 μM NMDA, 10 μM CNQX, and 2 μM nimodipine in medium containing 45CaCl2. CMP: counts per minute. * P < 0.05 vs corresponding control. n = 6 repeats.

Article Snippet: To examine whether NMDA-induced neurotoxicity was dependent on the NMDA receptor/NO signaling pathway, the cultured neurons were pre-incubated for 60 min with 10 μM MK-801 (an NMDA antagonist; Tocris, Ellisville, MI) or 30-60 μM N -Nitro-L-arginine methyl ester hydrochloride (L-NAME, a NOS inhibitor; Sigma, St. Louis, MO) at 37°C.

Techniques: Cell Culture, Control

Journal:

Article Title: PSD-93 Deficiency Protects Cultured Cortical Neurons from NMDA Receptor-triggered Neurotoxicity

doi: 10.1016/j.neuroscience.2010.01.030

Figure Lengend Snippet: PSD-93 deficiency decreases NMDA-stimulated increase in cGMP in cultured cortical neurons. (A) NMDA-induced neurotoxicity was NOS-dependent in cultured cortical neurons. Cultured neurons were treated with NMDA (30 μM or 60 μM) with or without L-NAME (10 μM or 30 μM). Neuronal viability was determined by MTT assay (top) and percentage of neuronal death by propidium iodide and calcein AM staining (bottom). * P < 0.05 vs NMDA alone. n = 6 repeats. (B) Cultured neurons from WT and PSD-93 KO mice were exposed to 30 μM NMDA. PSD-93 deficiency decreased the NMDA-stimulated increase in cGMP level in cultured cortical neurons. * P < 0.05 vs the corresponding NMDA-treated group from WT mice. n = 6 repeats.

Article Snippet: To examine whether NMDA-induced neurotoxicity was dependent on the NMDA receptor/NO signaling pathway, the cultured neurons were pre-incubated for 60 min with 10 μM MK-801 (an NMDA antagonist; Tocris, Ellisville, MI) or 30-60 μM N -Nitro-L-arginine methyl ester hydrochloride (L-NAME, a NOS inhibitor; Sigma, St. Louis, MO) at 37°C.

Techniques: Cell Culture, MTT Assay, Staining

Journal: International Journal of Molecular Sciences

Article Title: Fluctuating NMDA Receptor Subunit Levels in Perirhinal Cortex Relate to Their Dynamic Roles in Object Memory Destabilization and Reconsolidation

doi: 10.3390/ijms22010067

Figure Lengend Snippet: GluN2A, GluN2B, and GluN1 levels are significantly greater 24 h following learning compared to 48 h in PRh but not HPC. ( a ) Schematic of behavioral parameters. Rats in the learning group underwent a series of sample phases, then PRh and HPC tissues were collected either 24 or 48 h later. Non-learning controls remained in home cage. ( b ) Representative blot of GluN2A in anterior (A) and posterior (P) PRh in learning (+) and non-learning (-) groups 24 and 48 h following learning. GluN2A in whole PRh was significantly greater 24 h following learning compared to non-learning controls and 48 h following learning. ( c ) Representative blot of GluN2B in anterior and posterior PRh. GluN2B was significantly increased in whole PRh 24 h following learning compared to non-learning controls and 48 h following learning. ( d ) Representative blot of GluN1 in anterior and posterior PRh. GluN1 in PRh was significantly greater 24 vs 48 h following learning, but was not different from non-learning controls. ( e ) Representative blot of GluN2A in HPC. GluN2A levels did not differ across groups at 24 or 48 h following learning. ( f ) Representative blot of GluN2B in HPC. GluN2B levels did not differ across groups at 24 or 48 h following learning. ( g ) Representative blot of GluN1 in HPC. GluN1 levels did not differ across groups 24 or 48 h following learning. Bars represent mean GluN2 or GluN1 target–actin normalized to control ± SEM; * p < 0.05, ** p < 0.01.

Article Snippet: Conditions for each antibody were as follows: GluN2A (Millipore Sigma, Milwaukee, WI, USA, 07-632; blocking: 3% milk in TBS-T; primary: 1:1000 in 3% milk in TBS-T; secondary: anti-rabbit 1:2500 in 3% milk in TBS-T); GluN2B (Sigma-Aldrich, Oakville, ON, Canada, 06-600; blocking: 3% milk in PBS; primary: 1:1000 in 3% milk in PBS; secondary: anti-rabbit 1: 2500 in 3% milk in PBS); B-Actin (Cell Signaling Technology, Whitby, ON, Canada, 8H10D10; blocking: 5% BSA in 0.1% TBS-T; primary: 1:5000 in 5% BSA in 0.1% TBS-T; secondary: anti-mouse 1:2500 in 5% BSA in 0.1% TBS-T); GluN1 (Cell Signaling Technology; 5704S; blocking: 3% BSA in 0.1% TBS-T; primary 1:1000 in 3% BSA in 0.1% TBS-T; secondary: anti-mouse 1:2500 in 3% BSA in 0.1% TBS-T).

Techniques: Control