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a Schematic of the AAV used and experimental procedure for anterograde tracing. b Representative AAV-hSyn-mGFP-Syp-mRuby infection pattern in the LHb (inset) and the sagittal fluorescent image showing LHb D-neuron projections. c , A rotated and magnified image of the region of interest (dashed square) from ( b ). LHb D-neuronal terminals on GABAergic neurons of the RMTg were detected by FISH. d Schematic of AAV-ChR2 injection into the LHb and the response of RMTg GABAergic neurons to brief optical stimulation. e Traces from oEPSCs (left) and summary data on normalised oEPSC amplitude under baseline conditions after CNQX (20 μM) and <t>AP5</t> (50 μM) perfusion in ACSF (right) ( n = 9 cells from independent ChR2 mice). f Schematic of the method for measuring the firing rate of RMTg GABAergic neurons in AADC-knockdown VGAT Cre ::Ai14 mice and their vehicle controls. g Representative traces (left), firing frequency (middle; n = 9 cells from independent sh-Veh mice and n = 14 cells from independent sh-AADC mice) and RMP (right; n = 10 cells from independent sh-Veh mice and n = 14 cells from independent sh-AADC mice). Unless otherwise stated, statistical comparisons were performed using a two-tailed unpaired t test. * p < 0.05, ** p < 0.01 and *** p < 0.001. Data are presented as the mean values ± s.e.m. Source data are provided as a Source Data file.
Nmda Receptor Antagonist Ap5, supplied by Bio-Techne corporation, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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a Schematic of the AAV used and experimental procedure for anterograde tracing. b Representative AAV-hSyn-mGFP-Syp-mRuby infection pattern in the LHb (inset) and the sagittal fluorescent image showing LHb D-neuron projections. c , A rotated and magnified image of the region of interest (dashed square) from ( b ). LHb D-neuronal terminals on GABAergic neurons of the RMTg were detected by FISH. d Schematic of AAV-ChR2 injection into the LHb and the response of RMTg GABAergic neurons to brief optical stimulation. e Traces from oEPSCs (left) and summary data on normalised oEPSC amplitude under baseline conditions after CNQX (20 μM) and <t>AP5</t> (50 μM) perfusion in ACSF (right) ( n = 9 cells from independent ChR2 mice). f Schematic of the method for measuring the firing rate of RMTg GABAergic neurons in AADC-knockdown VGAT Cre ::Ai14 mice and their vehicle controls. g Representative traces (left), firing frequency (middle; n = 9 cells from independent sh-Veh mice and n = 14 cells from independent sh-AADC mice) and RMP (right; n = 10 cells from independent sh-Veh mice and n = 14 cells from independent sh-AADC mice). Unless otherwise stated, statistical comparisons were performed using a two-tailed unpaired t test. * p < 0.05, ** p < 0.01 and *** p < 0.001. Data are presented as the mean values ± s.e.m. Source data are provided as a Source Data file.
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a Schematic of the AAV used and experimental procedure for anterograde tracing. b Representative AAV-hSyn-mGFP-Syp-mRuby infection pattern in the LHb (inset) and the sagittal fluorescent image showing LHb D-neuron projections. c , A rotated and magnified image of the region of interest (dashed square) from ( b ). LHb D-neuronal terminals on GABAergic neurons of the RMTg were detected by FISH. d Schematic of AAV-ChR2 injection into the LHb and the response of RMTg GABAergic neurons to brief optical stimulation. e Traces from oEPSCs (left) and summary data on normalised oEPSC amplitude under baseline conditions after CNQX (20 μM) and <t>AP5</t> (50 μM) perfusion in ACSF (right) ( n = 9 cells from independent ChR2 mice). f Schematic of the method for measuring the firing rate of RMTg GABAergic neurons in AADC-knockdown VGAT Cre ::Ai14 mice and their vehicle controls. g Representative traces (left), firing frequency (middle; n = 9 cells from independent sh-Veh mice and n = 14 cells from independent sh-AADC mice) and RMP (right; n = 10 cells from independent sh-Veh mice and n = 14 cells from independent sh-AADC mice). Unless otherwise stated, statistical comparisons were performed using a two-tailed unpaired t test. * p < 0.05, ** p < 0.01 and *** p < 0.001. Data are presented as the mean values ± s.e.m. Source data are provided as a Source Data file.
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a Schematic of the AAV used and experimental procedure for anterograde tracing. b Representative AAV-hSyn-mGFP-Syp-mRuby infection pattern in the LHb (inset) and the sagittal fluorescent image showing LHb D-neuron projections. c , A rotated and magnified image of the region of interest (dashed square) from ( b ). LHb D-neuronal terminals on GABAergic neurons of the RMTg were detected by FISH. d Schematic of AAV-ChR2 injection into the LHb and the response of RMTg GABAergic neurons to brief optical stimulation. e Traces from oEPSCs (left) and summary data on normalised oEPSC amplitude under baseline conditions after CNQX (20 μM) and <t>AP5</t> (50 μM) perfusion in ACSF (right) ( n = 9 cells from independent ChR2 mice). f Schematic of the method for measuring the firing rate of RMTg GABAergic neurons in AADC-knockdown VGAT Cre ::Ai14 mice and their vehicle controls. g Representative traces (left), firing frequency (middle; n = 9 cells from independent sh-Veh mice and n = 14 cells from independent sh-AADC mice) and RMP (right; n = 10 cells from independent sh-Veh mice and n = 14 cells from independent sh-AADC mice). Unless otherwise stated, statistical comparisons were performed using a two-tailed unpaired t test. * p < 0.05, ** p < 0.01 and *** p < 0.001. Data are presented as the mean values ± s.e.m. Source data are provided as a Source Data file.
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a Schematic of the AAV used and experimental procedure for anterograde tracing. b Representative AAV-hSyn-mGFP-Syp-mRuby infection pattern in the LHb (inset) and the sagittal fluorescent image showing LHb D-neuron projections. c , A rotated and magnified image of the region of interest (dashed square) from ( b ). LHb D-neuronal terminals on GABAergic neurons of the RMTg were detected by FISH. d Schematic of AAV-ChR2 injection into the LHb and the response of RMTg GABAergic neurons to brief optical stimulation. e Traces from oEPSCs (left) and summary data on normalised oEPSC amplitude under baseline conditions after CNQX (20 μM) and <t>AP5</t> (50 μM) perfusion in ACSF (right) ( n = 9 cells from independent ChR2 mice). f Schematic of the method for measuring the firing rate of RMTg GABAergic neurons in AADC-knockdown VGAT Cre ::Ai14 mice and their vehicle controls. g Representative traces (left), firing frequency (middle; n = 9 cells from independent sh-Veh mice and n = 14 cells from independent sh-AADC mice) and RMP (right; n = 10 cells from independent sh-Veh mice and n = 14 cells from independent sh-AADC mice). Unless otherwise stated, statistical comparisons were performed using a two-tailed unpaired t test. * p < 0.05, ** p < 0.01 and *** p < 0.001. Data are presented as the mean values ± s.e.m. Source data are provided as a Source Data file.
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a Schematic of the AAV used and experimental procedure for anterograde tracing. b Representative AAV-hSyn-mGFP-Syp-mRuby infection pattern in the LHb (inset) and the sagittal fluorescent image showing LHb D-neuron projections. c , A rotated and magnified image of the region of interest (dashed square) from ( b ). LHb D-neuronal terminals on GABAergic neurons of the RMTg were detected by FISH. d Schematic of AAV-ChR2 injection into the LHb and the response of RMTg GABAergic neurons to brief optical stimulation. e Traces from oEPSCs (left) and summary data on normalised oEPSC amplitude under baseline conditions after CNQX (20 μM) and AP5 (50 μM) perfusion in ACSF (right) ( n = 9 cells from independent ChR2 mice). f Schematic of the method for measuring the firing rate of RMTg GABAergic neurons in AADC-knockdown VGAT Cre ::Ai14 mice and their vehicle controls. g Representative traces (left), firing frequency (middle; n = 9 cells from independent sh-Veh mice and n = 14 cells from independent sh-AADC mice) and RMP (right; n = 10 cells from independent sh-Veh mice and n = 14 cells from independent sh-AADC mice). Unless otherwise stated, statistical comparisons were performed using a two-tailed unpaired t test. * p < 0.05, ** p < 0.01 and *** p < 0.001. Data are presented as the mean values ± s.e.m. Source data are provided as a Source Data file.

Journal: Nature Communications

Article Title: Neural mechanism of acute stress regulation by trace aminergic signalling in the lateral habenula in male mice

doi: 10.1038/s41467-023-38180-7

Figure Lengend Snippet: a Schematic of the AAV used and experimental procedure for anterograde tracing. b Representative AAV-hSyn-mGFP-Syp-mRuby infection pattern in the LHb (inset) and the sagittal fluorescent image showing LHb D-neuron projections. c , A rotated and magnified image of the region of interest (dashed square) from ( b ). LHb D-neuronal terminals on GABAergic neurons of the RMTg were detected by FISH. d Schematic of AAV-ChR2 injection into the LHb and the response of RMTg GABAergic neurons to brief optical stimulation. e Traces from oEPSCs (left) and summary data on normalised oEPSC amplitude under baseline conditions after CNQX (20 μM) and AP5 (50 μM) perfusion in ACSF (right) ( n = 9 cells from independent ChR2 mice). f Schematic of the method for measuring the firing rate of RMTg GABAergic neurons in AADC-knockdown VGAT Cre ::Ai14 mice and their vehicle controls. g Representative traces (left), firing frequency (middle; n = 9 cells from independent sh-Veh mice and n = 14 cells from independent sh-AADC mice) and RMP (right; n = 10 cells from independent sh-Veh mice and n = 14 cells from independent sh-AADC mice). Unless otherwise stated, statistical comparisons were performed using a two-tailed unpaired t test. * p < 0.05, ** p < 0.01 and *** p < 0.001. Data are presented as the mean values ± s.e.m. Source data are provided as a Source Data file.

Article Snippet: To confirm the presence of glutamatergic inputs, optically evoked EPSCs (oEPSCs) were recorded during the application of the competitive AMPA/kainate receptor antagonist CNQX (20 μM; Bio-Techne Corporation, Minneapolis, MN, USA) and the selective NMDA receptor antagonist AP5 (50 μM; Bio-Techne Corporation).

Techniques: Anterograde Tracing, Infection, Injection, Two Tailed Test