Reagents were bath applied following dilution into ACSF from stock solutions prepared in water or DMSO. DCG-IV (2S,2′R,3′R-2-[2′,3′-dicarboxycyclopropyl]glycine), forskolin, and GYKI 53655 were purchased from Tocris Bioscience (Bristol, UK). Picrotoxin and all salts for making ACSF and internal solution were obtained from Sigma-Aldrich. The final DMSO concentration was <0.01% total volume.
Gyki 53655
Manufactured by Bio-Techne
Sourced in United Kingdom
GYKI 53655 is a chemical compound that acts as a selective antagonist of the AMPA subtype of glutamate receptors. It is commonly used in scientific research to study the role of AMPA receptors in various biological processes.
Automatically generated - may contain errors
Lab products found in correlation
L ap4, by Bio-Techne (3 mentions)
Dcg 4, by Bio-Techne (2 mentions)
D ap5, by Abcam (2 mentions)
Jnj 55511118, by Bio-Techne (2 mentions)
Mk 801, by Bio-Techne (2 mentions)
Forskolin, by Bio-Techne (1 mentions)
Bicuculline, by Enzo Life Sciences (1 mentions)
Naspm, by Alomone (1 mentions)
Kainate, by Abcam (1 mentions)
Iem 1460, by Bio-Techne (1 mentions)
Nimodipine, by Bio-Techne (1 mentions)
Cgp55845, by Bio-Techne (1 mentions)
Nahco3, by Merck Group (1 mentions)
Tetrodotoxin ttx, by Bio-Techne (1 mentions)
Perampanel, by Adooq (1 mentions)
11 protocols using gyki 53655
1
Pharmacological Modulation of Synaptic Activity
2
Pharmacological Agents in Perfusion
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Drugs were added to the perfusion solution. The following agents were used: L-(+)-2-amino-4-phophonobutyric acid (L-AP4; 20 μM; Tocris Bioscience, catalog #0103), D-(−)-2-amino-5-phophonopentanoic acid (D-AP5; 50 μM; Abcam Biochemicals, catalog #120003), 1-(4-aminophenyl)-3-methylcarbamyl-4-methyl-3,4-dihydro-7,8-meth- ylenedioxy-5H-2,3-benzodiazepine hydrochloride [GYKI-53655 (GYKI); 50 μM;Tocris Bioscience catalog #2555), (S)-1-(2-Amino-2-carboxyethyl)-3-(2-carboxy-5-phenylthiophene-3-yl-methyl)-5-methylpyrimidine-2,4-dione (ACET; 1 μM; Tocris Bioscience, catalog #2728).
3
Patch-Clamp Recordings of CA1 Pyramidal Neurons
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After resting in the recovery solution for at least 1 h, individual hippocampal slices were transferred to the recording chamber, which was constantly perfused at a flow rate of 3 ml/min with artificial cerebrospinal fluid (ACSF) containing (in mM) 2 CaCl2, 20 glucose, 4.5 KCl, 1 MgCl2, 125 NaCl, 26 NaHCO3, and 1.25 NaH2PO4 and gassed with 95% O2 and 5% CO2 to ensure oxygen saturation and to maintain a pH value of 7.4. 30 μM D‐AP5 (Abcam and Tocris, USA), 20 μM GYKI53655 (Tocris), and 10 μM bicuculline (Enzo, USA) were added to the ACSF to block NMDAR‐, AMPAR‐, and GABAAR‐mediated synaptic transmission. Somatic whole‐cell recordings from CA1 pyramidal neurons were performed with a borosilicate glass pipette with the resistance of ca. 7 MΩ filled with internal solution which contained (in mM) 148 K‐gluconate, 10 HEPES, 10 NaCl, 0.5 MgCl2, 4 Mg‐ATP, and 0.4 Na3‐GTP. The pH value of internal solution was adjusted to 7.3 with KOH. Voltage‐clamp measurements were carried out using an EPC9/2 patch‐clamp amplifier (HEKA, Germany). The membrane potential was held at −70 mV in voltage‐clamp mode without liquid junction potential adjustment. Data acquisition and the generation of stimulation protocols were applied by the use of PULSE software (HEKA). Data were collected at 10 kHz and Bessel‐filtered at 2.9 kHz and analyzed through Igor 5 software (Wavemetrics, USA).
4
Whole-cell Patch-clamp Recording of Freshly Isolated Cells
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Freshly isolated cells were obtained from slices after protease treatment as described (Matthias et al., 2003 (link)). K+ channel blockers (100 μM Ba2+, 100 μM quinine) as well as AMPA receptor agonists, blockers and modulators were applied by transferring the cells with a tube electrode to the different solutions (Seifert and Steinhäuser, 1995 (link)). Membrane currents were measured in the whole-cell configuration as described above. The resistance of the patch pipettes was 4 MΩ, the input resistance was determined as described above. The bath solution contained (in mM): 150 NaCl, 5 KCl, 2 MgCl2, 2 CaCl2, 10 HEPES, and 10 glucose (pH 7.4). The pipette solution was the same as for slice recordings. For recordings in high Ca2+ solution, 150 mM NaCl was replaced by 50 mM CaCl2 and adjusted with N-methyl-D -glucamine (NMDG) to an osmolarity of 320 mOsm (pH 7.4). As for recording synaptic currents (cf. above), CsCl-based pipette solution was used. Recordings were obtained at room temperature. Salts and buffers were purchased from AppliChem (Darmstadt, Germany), kainate and CTZ were received from Abcam (Milton, United Kingdom), Naspm from Alomone Labs (Israel), and IEM-1460, JNJ 55511118 and GYKI53655 from Tocris (Bristol, United Kingdom).
5
Neurochemical Agents in Neuroscience
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MK-801, NBQX, CGP-55845, nimodipine, DCG-IV, and GYKI 53655 were obtained from Tocris-Cookson. LY 303070 was obtained from ABX advanced biochemical compounds. BoTX was obtained from List Biological. All other chemicals and drugs were purchased from Sigma-Aldrich.
6
Pharmacological Agents in Perfusion
7
Retinal Dissection and Electrophysiological Recording
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The retinal dissections were performed in HEPES-buffered extracellular solution containing the following (in mM):115 NaCl, 2.5 KCl, 2.5 CaCl2, 1.0 MgCl2,10 HEPES, and 28 glucose, adjusted to pH 7.37 with NaOH. Physiological recordings were performed in Ames’ medium buffered with NaHCO3 (Millipore-Sigma) and bubbled with 95% O2 and 5% CO2; the pH was 7.4 at 32 – 34°C. The intracellular solution contained the following (in mM):110 potassium methylsulfonate, 10 HEPES, 4 EGTA, 5 NaCl, 5 KCl, 1 MgCl2, 4 ATP-Mg, and 1 GTP-Na, adjusted to pH 7.2 with KOH. For voltage-clamp recordings, the intracellular solution contained the following: 110 cesium methylsulfonate, 10 HEPES, 10 TEA-Cl, 4 EGTA, 1 MgCl2, 5 mM QX-314, 4 ATP-Mg, and 1 GTP-Na, adjusted to pH 7.2 with CsOH. To block photoreceptor inputs to bipolar cells and SACs, 10 μM L-AP4 (Tocris), 1 μM ACET (Tocris), and 50 μM GYKI53655 (Tocris, Bristol, UK) were perfused in the bath solution.
8
Kainate-induced Neuronal Excitation
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Rat cortical neurons were plated in a 6-well dish with a density of 500,000 cells per well. The cells were incubated at 37°C until reaching a developmental stage of DIV 19–21 DIV. On the experiment day, the cells were subjected to pre-treatment by the addition of 2 μM tetrodotoxin (TTX, a Na2+ channel blocker; Tocris) and 40 μM GYKI 53655 (a selective AMPAR blocker; Tocris) to the culture medium for 30 min. Following pre-treatment, KA incubation (10 μM for 20 min at 37°C) was performed in the continued presence of TTX and GYKI 53655. Control cells were treated with a vehicle instead of KA.
9
Antagonists Modulate Neuronal Excitability
10
Kainate-induced excitotoxicity in hippocampal neurons
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Hippocampal neurons were plated in a 6 well culture dish (500,000 cells per well) and maintained in a 37°C incubator. On DIV 17-18, the neurons were pre-treated with 1μM TTX (Tocris) and GYKI 53655 (Tocris), with or without additional drugs in Earle’s Buffer (140mM NaCl, 5mM KCl, 1.8mM CaCl2, 0.8mM MgCl2, 25mM HEPES, 0.9g/L D-Glucose, pH 7.4) and placed back in the incubator for 30 mins. After the incubation, 10μM KA was added to the wells and incubated for 20 mins at 37°C.
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