Journal: Antioxidants

Article Title: The Application of the Neuroprotective and Potential Antioxidant Effect of Ergotamine Mediated by Targeting N-Methyl-D-Aspartate Receptors

doi: 10.3390/antiox11081471

Figure Lengend Snippet: ( A ) The chemical structure of ergotamine. ( B ) The H 2 O-injected oocytes did not cause any change with the treatment of 100 µM glutamate ( n = 6–8 oocytes from four different frogs). ( C – F ) Glutamate induced inward currents with or without ergotamine (30 µM and 10 µM). For each subunit of NMDARs, the responses after treating with either glutamate (100 µM) alone or together with ergotamine (30 and 10 µM). Voltage clamp recording was conducted at a holding potential of −80 mV. The coapplication of ergotamine with glutamate resulted in modulation of the recombinant receptors ( C ) NR1a/NR2A, ( D ) NR1a/NR2B, ( E ) NR1a/NR2C, and ( F ) NR1a/NR2D, which in turn reduced glutamate-evoked inward current in a reversible manner ( n = 6–8 oocytes from four different frogs).

Article Snippet: The mouse NMDAR subunit cDNAs included the NR1 (GenBank accession number: MR225704), NR2A (MR227135), NR2B (MR227077), NR2C (MR222676), and NR2D (MR220972) subunits, which were purchased from OriGene (Rockville, MD, USA).

Techniques: Injection, Recombinant

Journal: Antioxidants

Article Title: The Application of the Neuroprotective and Potential Antioxidant Effect of Ergotamine Mediated by Targeting N-Methyl-D-Aspartate Receptors

doi: 10.3390/antiox11081471

Figure Lengend Snippet: The value of I max , IC 50 , and n H (Hill coefficient) of ergotamine for the glutamate-evoked current in each recombinant receptor. Values represent means ± S.E.M. ( n = 6–8/group). IC 50 , Hill’s coefficient; I max value as determined as described in Materials and methods.

Article Snippet: The mouse NMDAR subunit cDNAs included the NR1 (GenBank accession number: MR225704), NR2A (MR227135), NR2B (MR227077), NR2C (MR222676), and NR2D (MR220972) subunits, which were purchased from OriGene (Rockville, MD, USA).

Techniques: Recombinant

Journal: Antioxidants

Article Title: The Application of the Neuroprotective and Potential Antioxidant Effect of Ergotamine Mediated by Targeting N-Methyl-D-Aspartate Receptors

doi: 10.3390/antiox11081471

Figure Lengend Snippet: The predicted docking sites and binding energy of NR1a/NR2A receptor and ergotamine.

Article Snippet: The mouse NMDAR subunit cDNAs included the NR1 (GenBank accession number: MR225704), NR2A (MR227135), NR2B (MR227077), NR2C (MR222676), and NR2D (MR220972) subunits, which were purchased from OriGene (Rockville, MD, USA).

Techniques: Binding Assay

Journal: Antioxidants

Article Title: The Application of the Neuroprotective and Potential Antioxidant Effect of Ergotamine Mediated by Targeting N-Methyl-D-Aspartate Receptors

doi: 10.3390/antiox11081471

Figure Lengend Snippet: Computational molecular modeling of ergotamine docked to the human NR1a/NR2A receptor. ( A , C ) Side views of the docked ergotamine complex with NMDA channel. ( B , D ) Binding pocket and docking results of ergotamine and NMDA channel, respectively.

Article Snippet: The mouse NMDAR subunit cDNAs included the NR1 (GenBank accession number: MR225704), NR2A (MR227135), NR2B (MR227077), NR2C (MR222676), and NR2D (MR220972) subunits, which were purchased from OriGene (Rockville, MD, USA).

Techniques: Binding Assay

Journal: Antioxidants

Article Title: The Application of the Neuroprotective and Potential Antioxidant Effect of Ergotamine Mediated by Targeting N-Methyl-D-Aspartate Receptors

doi: 10.3390/antiox11081471

Figure Lengend Snippet: Predicted binding mode of ergotamine and all the favorable interactions with several residues in the active site of the human NR1a/NR2A receptor. ( A , B ) Interaction between ergotamine and wild-type NR1a/NR2A. ( C ) Residues in wild-type NR1a/NR2A receptor interacting with the ergotamine molecule. ( D ) Change in the interaction distance of ergotamine in mutant-type NR1a/NR2A receptor. Based on the change in this distance, the residue that directly interacts with ergotamine was identified.

Article Snippet: The mouse NMDAR subunit cDNAs included the NR1 (GenBank accession number: MR225704), NR2A (MR227135), NR2B (MR227077), NR2C (MR222676), and NR2D (MR220972) subunits, which were purchased from OriGene (Rockville, MD, USA).

Techniques: Binding Assay, Mutagenesis

Journal: Antioxidants

Article Title: The Application of the Neuroprotective and Potential Antioxidant Effect of Ergotamine Mediated by Targeting N-Methyl-D-Aspartate Receptors

doi: 10.3390/antiox11081471

Figure Lengend Snippet: Effect of ergotamine on the glutamate-evoked current of wild-type NR1a/NR2A receptor and its various mutants.

Article Snippet: The mouse NMDAR subunit cDNAs included the NR1 (GenBank accession number: MR225704), NR2A (MR227135), NR2B (MR227077), NR2C (MR222676), and NR2D (MR220972) subunits, which were purchased from OriGene (Rockville, MD, USA).

Techniques:

Journal: Antioxidants

Article Title: The Application of the Neuroprotective and Potential Antioxidant Effect of Ergotamine Mediated by Targeting N-Methyl-D-Aspartate Receptors

doi: 10.3390/antiox11081471

Figure Lengend Snippet: The inward current of several mutant types on the glutamate-evoked current of NR1a/NR2A receptor with or without ergotamine. Representative traces of current induced by application of glutamate (100 µM) alone or together with ergotamine (100 µM) for various mutants: ( A ) mutant 1 (NR1a subunit W167A and NR2A wild-type), ( B ) mutant 2 (NR1a subunit H168A and NR2A wild-type), ( C ) mutant 3 (NR1a subunit V169A and NR2A wild-type), ( D ) mutant 4 (NR1a wild-type and NR2A subunit P435A), ( E ) mutant 5 (NR1a wild-type and NR2A subunit N466A), and ( F ) mutant 6 (double mutant-type NR1a subunit V169A and NR2A subunit N466A). Experiments were performed separately, and data were collected from several oocytes ( n = 6–8 oocytes from four different frogs).

Article Snippet: The mouse NMDAR subunit cDNAs included the NR1 (GenBank accession number: MR225704), NR2A (MR227135), NR2B (MR227077), NR2C (MR222676), and NR2D (MR220972) subunits, which were purchased from OriGene (Rockville, MD, USA).

Techniques: Mutagenesis

Journal: Antioxidants

Article Title: The Application of the Neuroprotective and Potential Antioxidant Effect of Ergotamine Mediated by Targeting N-Methyl-D-Aspartate Receptors

doi: 10.3390/antiox11081471

Figure Lengend Snippet: ( A ) Concentration-response curves for the effect of ergotamine on expressed NMDA receptors. The percentage inhibition by ergotamine was calculated based on the average of peak inward current elicited by glutamate and that of peak inward current elicited by glutamate and ergotamine. Each point represents the mean ± S.E.M. ( n = 6–8 oocytes from four different frogs). ( B ) Concentration–response curves for the inhibition of ergotamine on glutamate-induced inward current of NR1a/NR2A mutant types. NR1a (W167A, H168A, and V169A) and NR2A mutants (P435A and N466A) were cross-combined between wild-type/mutants of each subunit. Ergotamine reduced I Glu in a concentration-dependent manner in the wild-type. Experiments were performed at a holding potential of −80 mV. The value of IC 50 , I max , and n H (Hill coefficient) are shown in . Each point represents the mean ± S.E.M. ( n = 6–8 oocytes from four different frogs).

Article Snippet: The mouse NMDAR subunit cDNAs included the NR1 (GenBank accession number: MR225704), NR2A (MR227135), NR2B (MR227077), NR2C (MR222676), and NR2D (MR220972) subunits, which were purchased from OriGene (Rockville, MD, USA).

Techniques: Concentration Assay, Inhibition, Mutagenesis

Journal: Scientific Reports

Article Title: Running in mice increases the expression of brain hemoglobin-related genes interacting with the GH/IGF-1 system

doi: 10.1038/s41598-024-77489-1

Figure Lengend Snippet: Information on transcripts.

Article Snippet: Grin2a , Mm00433802_m1 , Glutamate receptor, ionotropic, 2a , Neuron , Brain plasticity.

Techniques: Functional Assay, Membrane, Knock-In, Transmission Assay

Journal: Scientific Reports

Article Title: Running in mice increases the expression of brain hemoglobin-related genes interacting with the GH/IGF-1 system

doi: 10.1038/s41598-024-77489-1

Figure Lengend Snippet: Quantitative reverse transcription polymerase chain reaction (RT-qPCR) values in runners and sedentary animals.

Article Snippet: Grin2a , Mm00433802_m1 , Glutamate receptor, ionotropic, 2a , Neuron , Brain plasticity.

Techniques: Reverse Transcription, Polymerase Chain Reaction

Journal: PLoS ONE

Article Title: MPX-004 and MPX-007: New Pharmacological Tools to Study the Physiology of NMDA Receptors Containing the GluN2A Subunit

doi: 10.1371/journal.pone.0148129

Figure Lengend Snippet: The values are IC 50 s for inhibition of Ca 2+ responses mediated by GluN2A receptors expressed in HEK cells.

Article Snippet: The human GRIN1 gene transcript variant NR1-3 (Origene, SKU#: SC115601, Rockville, MD), GRIN2A gene (Origene, SKU#: SC326381), GRIN2B gene (B’SYS, Witterswil, Switzerland), GRIN2C gene (NCBI locus NM_000835, optimized for oocyte expression and synthesized by GenScript, Piscataway, NJ) and GRIN2D gene (NCBI locus NM_000836, sequence optimized and synthesized by GenScript) were linearized and transcribed using the mMessage mMachine kit (Life Technologies) and promoters SP6 (for GluN1) and T7 (for GluN2A-2D). cRNAs were mixed together at a ratio of 1 GluN1:2 GluN2.

Techniques: Inhibition

Journal: PLoS ONE

Article Title: MPX-004 and MPX-007: New Pharmacological Tools to Study the Physiology of NMDA Receptors Containing the GluN2A Subunit

doi: 10.1371/journal.pone.0148129

Figure Lengend Snippet: Cells were stimulated with glutamate and glycine (3 μM each) in the presence of compounds at a range of concentrations. Curves for inhibition of the Ca 2+ response in GluN2A-expressing cells were derived from fits to the Hill equation using GraphPad Prism (v6.00 for Mac, GraphPad Software, La Jolla California USA). Whereas MPX-004 and MPX-007 achieve full inhibition of the GluN2A Ca 2+ response by ~ 3 μM, TCN-201 never inhibits more than ~40% of the response. Each data point is a mean (± standard deviation) of data from 20–86 experiments).

Article Snippet: The human GRIN1 gene transcript variant NR1-3 (Origene, SKU#: SC115601, Rockville, MD), GRIN2A gene (Origene, SKU#: SC326381), GRIN2B gene (B’SYS, Witterswil, Switzerland), GRIN2C gene (NCBI locus NM_000835, optimized for oocyte expression and synthesized by GenScript, Piscataway, NJ) and GRIN2D gene (NCBI locus NM_000836, sequence optimized and synthesized by GenScript) were linearized and transcribed using the mMessage mMachine kit (Life Technologies) and promoters SP6 (for GluN1) and T7 (for GluN2A-2D). cRNAs were mixed together at a ratio of 1 GluN1:2 GluN2.

Techniques: Inhibition, Expressing, Derivative Assay, Software, Standard Deviation

Journal: PLoS ONE

Article Title: MPX-004 and MPX-007: New Pharmacological Tools to Study the Physiology of NMDA Receptors Containing the GluN2A Subunit

doi: 10.1371/journal.pone.0148129

Figure Lengend Snippet: C ells were stimulated with glutamate and glycine (3 μM each) in the presence of compounds at a range of concentrations. IC 50 for inhibition of the Ca 2+ response in GluN2A-expressing cells was fitted to the Hill equation using CDD Vault. For GluN2B or GluN2D no IC 50 could be determined, so the effect of each compound at 10 μM is shown as the % inhibition of the Ca 2+ response (note that negative % inhibition represents an increase in Ca 2+ response over glutamate plus glycine alone). Values are the mean IC 50 or % ± SEM with the number of replicate curves indicated in parentheses. ND- not determined.

Article Snippet: The human GRIN1 gene transcript variant NR1-3 (Origene, SKU#: SC115601, Rockville, MD), GRIN2A gene (Origene, SKU#: SC326381), GRIN2B gene (B’SYS, Witterswil, Switzerland), GRIN2C gene (NCBI locus NM_000835, optimized for oocyte expression and synthesized by GenScript, Piscataway, NJ) and GRIN2D gene (NCBI locus NM_000836, sequence optimized and synthesized by GenScript) were linearized and transcribed using the mMessage mMachine kit (Life Technologies) and promoters SP6 (for GluN1) and T7 (for GluN2A-2D). cRNAs were mixed together at a ratio of 1 GluN1:2 GluN2.

Techniques: Inhibition, Expressing

Journal: PLoS ONE

Article Title: MPX-004 and MPX-007: New Pharmacological Tools to Study the Physiology of NMDA Receptors Containing the GluN2A Subunit

doi: 10.1371/journal.pone.0148129

Figure Lengend Snippet: Oocytes were exposed to MPX-004 or MPX-007 at concentrations from 10 nM to 10 μM as indicated. Inhibition curves were generated using GraphPad Prism and IC 50 values were generated using CCD Vault. The IC 50 s for inhibition of GluN2A-mediated currents were 198 ± 17 and 143 ± 10 nM for MPX-004 and MPX-007, respectively. Each data point is a mean (± standard deviation) of data from 4–12 oocytes.

Article Snippet: The human GRIN1 gene transcript variant NR1-3 (Origene, SKU#: SC115601, Rockville, MD), GRIN2A gene (Origene, SKU#: SC326381), GRIN2B gene (B’SYS, Witterswil, Switzerland), GRIN2C gene (NCBI locus NM_000835, optimized for oocyte expression and synthesized by GenScript, Piscataway, NJ) and GRIN2D gene (NCBI locus NM_000836, sequence optimized and synthesized by GenScript) were linearized and transcribed using the mMessage mMachine kit (Life Technologies) and promoters SP6 (for GluN1) and T7 (for GluN2A-2D). cRNAs were mixed together at a ratio of 1 GluN1:2 GluN2.

Techniques: Inhibition, Generated, Standard Deviation

Journal: PLoS ONE

Article Title: MPX-004 and MPX-007: New Pharmacological Tools to Study the Physiology of NMDA Receptors Containing the GluN2A Subunit

doi: 10.1371/journal.pone.0148129

Figure Lengend Snippet: Effect of MPX-004 on the ratio of NMDA receptor- to AMPA receptor-mediated synaptic currents (NMDAR/AMPAR ratio) at layer 4-to-2/3 synapses in mouse visual cortex.

Article Snippet: The human GRIN1 gene transcript variant NR1-3 (Origene, SKU#: SC115601, Rockville, MD), GRIN2A gene (Origene, SKU#: SC326381), GRIN2B gene (B’SYS, Witterswil, Switzerland), GRIN2C gene (NCBI locus NM_000835, optimized for oocyte expression and synthesized by GenScript, Piscataway, NJ) and GRIN2D gene (NCBI locus NM_000836, sequence optimized and synthesized by GenScript) were linearized and transcribed using the mMessage mMachine kit (Life Technologies) and promoters SP6 (for GluN1) and T7 (for GluN2A-2D). cRNAs were mixed together at a ratio of 1 GluN1:2 GluN2.

Techniques: