For electrophysiological recordings, 350-µm-thick brain slices were prepared from 3 week old mice and equilibrated in artificial cerebrospinal fluid (aCSF) composed of 120 mM NaCl, 3 mM KCl, 1.3 mM Mg2SO4, 1.25 mM NaH2PO4, 2.5 mM CaCl2, 10 mM D-glucose, and 25 mM NaHCO3, and gassed with 95% O2/5% CO2, pH 7.3 at room temperature for at least 1 h as described previously [34 (link)].
Patch clamp recordings were performed on coronal slices that were placed in a submerged recording chamber mounted on an upright microscope (BX51WI, Olympus, Hamburg, Germany). Slices were continuously superfused with gassed aCSF (2–3 mL/min, 32 °C, pH 7.3). Recordings of miniature inhibitory postsynaptic current (mIPSC) kinetics were performed using a CsCl-based intracellular solution containing 122 mM CsCl, 8 mM NaCl, 0.2 mM MgCl2, 10 mM HEPES, 2 mM EGTA, 2 MM Mg-ATP, 0.5 mM Na-GTP, 10 mM QX-314 [N-(2,6-dimethylphenylcarbamoylmethyl) triethylammonium bromide], pH adjusted to 7.3 with CsOH. DL-AP5 (30 μM), CNQX (10 μM) and tetrodotoxin (0.5 μM) were added to the perfusate. mIPSCs were recorded at a holding potential of −70 mV for at least 5 min in aCSF. Data analysis was performed off-line with the detection threshold levels set to 5 pA. The following parameters were determined: frequency, peak amplitude, rise time, time constant of decay (τ-decay), half-width, and electrical charge transfer.
Patch clamp recordings were performed on coronal slices that were placed in a submerged recording chamber mounted on an upright microscope (BX51WI, Olympus, Hamburg, Germany). Slices were continuously superfused with gassed aCSF (2–3 mL/min, 32 °C, pH 7.3). Recordings of miniature inhibitory postsynaptic current (mIPSC) kinetics were performed using a CsCl-based intracellular solution containing 122 mM CsCl, 8 mM NaCl, 0.2 mM MgCl2, 10 mM HEPES, 2 mM EGTA, 2 MM Mg-ATP, 0.5 mM Na-GTP, 10 mM QX-314 [N-(2,6-dimethylphenylcarbamoylmethyl) triethylammonium bromide], pH adjusted to 7.3 with CsOH. DL-AP5 (30 μM), CNQX (10 μM) and tetrodotoxin (0.5 μM) were added to the perfusate. mIPSCs were recorded at a holding potential of −70 mV for at least 5 min in aCSF. Data analysis was performed off-line with the detection threshold levels set to 5 pA. The following parameters were determined: frequency, peak amplitude, rise time, time constant of decay (τ-decay), half-width, and electrical charge transfer.
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