Cyclotraxin B and 6,7-dinitroquinoxaline-2,3-dione (DNQX) were purchased from Tocris (Minneapolis MN). Rapamycin was from Cayman Chemical (Ann Arbor MI). Ketamine, CNQX other agents were obtained from Millipore Sigma (St. Louis MO). N2O with 5.04% carbon dioxide was purchased from Puritan Medical Products (Overland KS) and was bubbled into a reservoir containing ACSF at the time of experiments (25 (link)). We measured N2O in ACSF as a way to monitor our solutions (Supplemental Methods and Results ). In some experiments, we administered 30% N2O in vivo prior to slice preparation. Because of use of different gas conditions and recording paradigms, experimenters were not blind to recording conditions. Experiments were done with 30% nitrous oxide, a maximal concentration for use in ex vivo slices (25 (link)).
6 7 dinitroquinoxaline 2 3 dione dnqx
Manufactured by Bio-Techne
Sourced in United Kingdom
6,7-dinitroquinoxaline-2,3-dione (DNQX) is a chemical compound used in laboratory research. It functions as a selective antagonist for the AMPA and kainate subtypes of ionotropic glutamate receptors.
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Lab products found in correlation
Cgp 55845a, by Bio-Techne (2 mentions)
Dl ap5, by Bio-Techne (2 mentions)
Picrotoxin, by Merck Group (2 mentions)
Rapamycin, by Cayman Chemical (1 mentions)
Cyclotraxin b, by Bio-Techne (1 mentions)
Taurine tau, by Merck Group (1 mentions)
Vybrant apoptosis assay kit, by Thermo Fisher Scientific (1 mentions)
Mk 801, by Bio-Techne (1 mentions)
Hoechst 33342, by Thermo Fisher Scientific (1 mentions)
Calcein am, by Merck Group (1 mentions)
Cell culture reagents, by Thermo Fisher Scientific (1 mentions)
Staurosporine, by Merck Group (1 mentions)
Picrotoxin, by Bio-Techne (1 mentions)
Dl ap4, by Bio-Techne (1 mentions)
1 2 5 6 tetrahydropyridin 4 yl methylphosphinic acid tpmpa, by Bio-Techne (1 mentions)
D 2 amino 5 phosphonopentanoic acid d apv, by Bio-Techne (1 mentions)
Tetraethyl ammonium tea, by Merck Group (1 mentions)
Tiagabine, by Merck Group (1 mentions)
Topiramate, by Merck Group (1 mentions)
Ethosuximide, by Merck Group (1 mentions)
9 protocols using 6 7 dinitroquinoxaline 2 3 dione dnqx
1
Nitrous Oxide and Neuroprotective Agents
2
Pharmacological Agents for Synaptic Studies
3
Apoptosis Assay in Cell Culture
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Cell culture reagents were obtained from Gibco (Grand Island, NY). Calcein AM was obtained from Sigma (St. Louis, MO). A Vybrant® Apoptosis Assay Kit (V13241) and Hoechst 33342 were purchased from Molecular Probes (Eugene, OR). A cell-dissection kit of Papain was obtained from Worthington Biochemical (Lakewood, NJ). Recombinant neurotrophic factors (human brain-derived neurotrophic factor [BDNF] and rat ciliary neurotrophic factor [CNTF]) were obtained from Sigma. Monoclonal ascites IgG2a antibody (OX-41) against rat macrophage and monoclonal IgG1 antibody (OX-7) against rat and mouse Thy-1.1 were obtained from Chemicon (Temecula, CA). Staurosporine was purchased from Sigma. The AMPA-KA receptor antagonist, 6,7-dinitroquinoxaline-2,3-dione (DNQX), and the NMDA receptor antagonist, MK 801, were obtained from Tocris Bioscience (Ellisville, MO).
4
Synaptic Blockade in Retina Explant
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Blocking of various synapses was achieved by bath applying various pharmacological substances dissolved in aCSF, which superfused the explanted retina as described. Gap junctions were blocked with 50 µM MFA (Sigma Aldrich) that was applied for 30 min before the data were recorded. Glutamatergic signaling was blocked using a combination of three compounds: 100 µM 6,7-dinitroquinoxaline-2,3-dione (DNQX; Tocris Bioscience, Bristol, UK) was used to block the α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA)-type glutamatergic synapses, 100 µM DL-AP5 (Tocris Bioscience) was used to block the N-methyl-D-aspartate (NMDA) glutamatergic synapses, and 150 µM DL-AP4 (Tocris Bioscience) was used to block the metabotropic (mGluR6) synapses. As an mGluR6 agonist, DL-AP4 did not allow the investigation of synapses downstream of the transfected ON bipolar cells. Gamma-aminobutyric acid (GABA)a and GABAc receptors were antagonized with 20 µM picrotoxin and 50 µM 1,2,5,6-tetrahydropyridin-4-yl)methylphosphinic acid (TPMPA), respectively (Tocris Bioscience).
5
Native α-LTX Toxin Preparation
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Lyophilized native α-LTX (>98% purity) was obtained from Alomone Labs (Jerusalem, Israel, LSP-130). 6,7-Dinitroquinoxaline-2,3-dione (DNQX, an AMPA receptor antagonist) and D(−)-2-amino-5-phosphonopentanoic acid (D-APV, an NMDA receptor antagonist) were obtained from Tocris Bioscience. All other chemicals used for preparation of the artificial cerebrospinal fluid (ACSF) and pipette solutions were obtained from Sigma-Aldrich.
6
Purchasing Specific Pharmacological Agents
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6,7‐dinitroquinoxaline‐2,3‐dione (DNQX) and (DL)‐amino‐5‐phosphonovaleric acid (APV), were purchased from Tocris Bioscience, (Bristol, UK). Picrotoxin, tetraethylammonium (TEA), and salts were purchased from Sigma–Aldrich (Milwaukee, WI).
7
Preparing Drug Stock Solutions
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Stock solutions of drugs were freshly prepared and filtered using 0.22 μM solvent resistant filters (Millipore) as follows: topiramate, picrotoxin and tiagabine (Sigma-Aldrich), retigabine, XE991 (both from Alamone) and kynurenic acid (Tocris) were dissolved in DMSO to obtain stock solutions of 10 mM. Bicuculline, d -2-Amino-5-phosphonovaleric acid (D-AP5), 4-Aminopyridine (4-AP), phenobarbital, valproic acid (all from Sigma-Aldrich), 6,7-dinitroquinoxaline-2,3-dione (DNQX), tetrodotoxin (TTX) (both from Tocris) were dissolved in water to obtain stock solutions of 10 mM. Carbamazepine, ethosuximide and diazepam (all from Sigma-Aldrich) were dissolved in ethanol to obtain stock solutions of 10 mM. All drugs were then serially diluted to their final concentrations in cell culture/recording media.
8
Preparation of Pharmacological Solutions
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Most drugs were prepared as concentrated stock solutions. 6,7‐dinitroquinoxaline‐2,3‐dione (DNQX 30 mmol/L; Tocris) was dissolved in dimethyl sulfoxide (DMSO), while CGP‐37849 (5 mmol/L, Tocris) bicuculline methiodide (10 mmol/L, Tocris) and 4‐AP (100 μmol/L, Sigma) were dissolved in water. Stock solutions were diluted to final concentrations by addition to ACSF perfusing the tissue. TEA (10 mmol/L; chloride salt) was dissolved directly into ACSF, substituted on an equimolar basis for NaCl. Salts and all other reagents were from Sigma or Fisher.
9
Behavioral and Electrophysiology Protocols for Semaglutide
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For behavioral testing, semaglutide (Peptide International) was prepared using 1.25% (v/v) dimethyl sulfoxide (DMSO; Thermo Fisher Scientific) and 1.25% (v/v) Tween 80 (Thermo Fisher Scientific), and diluted with 0.9% saline (Hospira). Following a within-subjects, Latin-square design, semaglutide (0.001, 0.003, 0.01, 0.03, 0.1 mg/kg in mice; 0.001, 0.01, 0.1 mg/kg in rats), and vehicle were administered s.c. The volume of injection was 10 mL/kg in mice and 1 mL/kg in rats. The alcohol solution used for systemic injections in the rotarod experiments in mice was prepared with 200-proof ethanol (Pharmco) in 0.9% saline to produce a 20% (v/v) alcohol solution. This solution was administered i.p. at a dose of 2.0 g/kg. For electrophysiology studies, stock solutions of semaglutide (BOC Sciences), CGP55845A (Tocris), 6,7-dinitroquinoxaline-2,3-dione (DNQX; Tocris), and DL-2-amino-5-phosphonovalerate (DL-AP5; Tocris) were prepared in either distilled water or DMSO, aliquoted, frozen, and added to the bath solution.
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