Chemicals inhibiting specific ion channels were introduced in the bath/recording buffer to compare different physiological components possibly contributing to the neuronal hyperactivity. To block all fast-synaptic excitatory transmission, 3-[(±)2-carboxypiperazin-4yl] propyl-1-phosphate (CPP, NMDA receptor antagonist, 20 μM; Tocris Bioscience, United Kingdom) and 6-cyano-7-nitroquinoxaline-2,3-dione [CNQX, AMPA/kainate (non-NMDA) receptor antagonist, 20 μM, diluted in DMSO; Alomone Labs, Israel] were introduced in the bath. SR95531 hydrobromide or Gabazine (GABAA receptor antagonist, 10 μM, Tocris Bioscience, United Kingdom) was used to block inhibitory synaptic transmission. Cd2+ (CdCl2, 100 μM; Sigma-Aldrich, United States) was used to depress synaptic transmission and block inward calcium-selective current, isolating the outward K+ current. To block voltage-dependent K+ channels, intracellular administration of cesium was done by replacing potassium gluconate with cesium gluconate in the internal solution while maintaining the same osmolarity.
6 cyano 7 nitroquinoxaline 2 3 dione
Manufactured by Alomone
Sourced in United Kingdom
6-cyano-7-nitroquinoxaline-2,3-dione is a chemical compound that functions as a selective antagonist of AMPA-type glutamate receptors.
Automatically generated - may contain errors
3 protocols using 6 cyano 7 nitroquinoxaline 2 3 dione
1
Dissecting Neuronal Hyperactivity Mechanisms
2
Glutamate Uncaging Reveals AMPAR and NMDAR Currents
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Neurons in the voltage-clamp mode were subjected to glutamate uncaging in the presence of TTX (0.5 μM) and picrotoxin (100 μM). Laser power was adjusted to 12 mW and the two-photon pulse duration was 0.72 ms. The membrane potential was maintained at −80 mV (after correction for the liquid junction potential) for the AMPA-EPSC measurement, and NMDA-EPSCs were measured at a holding potential of +40 mV with the AMPAR antagonist 6-cyano-7-nitroquinoxaline-2,3-dione (50 μM; Alomone Labs).
3
Patch-Clamp Analysis of Inhibitory Postsynaptic Currents
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We patch-clamped 10–17 d in vitro mouse neurons. Internal pipette solution contained (mM): 115 Cs-methanesulfonate, 10 CsCl, 5 NaCl, 10 HEPES, 20 tetraethylammonium chloride, 4 Mg-ATP, 0.3 NaGTP, 0.6 EGTA, and 10 lidocaine N-ethyl bromide, pH 7.2 with CsOH. The external solution used was the high sodium solution described above, containing 10 µM 6-cyano-7-nitroquinoxaline-2,3-dione (Alomone Labs) to isolate inhibitory postsynaptic currents (IPSCs). To elicit evoked responses, we delivered electrical stimulation through parallel platinum electrodes (1-ms duration; amplitude, 20 mA) while neurons were held at −80 mV. We added 1 µM TTX to record miniature IPSCs (mIPSCs).
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