Slices were perfused with ACSF recording solution supplemented with 1 μM tetrodotoxin to prevent action potentials and 100 μM picrotoxin to suppress inhibitory currents35.
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After at least 20 min of stable recording, the recording solution was switched to recording solution containing 0.01% DMSO; 15 min later hippocampal slices were perfused with recording solution containing 0.01% DMSO and either E2 or G1 at 10 nM.
Menthol and icilin stocks were dissolved in DMSO and diluted into recording solution immediately prior to experiments.
A recycling perfusion system was used to minimize recording solution volumes (5 mL) and allow the continuous perfusion of the caged MNI-glutamate (1 mM).
Standard recording solution contained (in mM): 90 KCl, 3 MgCl2, 10 Hepes (pH 7.4).
The offset potential of the tip was routinely adjusted after immersion into recording solution.
The recording solution contained (in mmol/L) 90 NaCl, 1 KCl, 10 HEPES, 0.5 BaCl2, 0.01 EDTA (pH 7.4).
Cells were continuously superfused in recording solution consisting of (in mmol/L): NaCl, 124; KCl, 3; NaHCO3, 26; NaH2PO4, 1.25; CaCl2, 2; MgSO4, 1; glucose, 10.
External recording solution contained (in mM): 114 CsCl, 20 BaCl2, 1 MgCl2, 10 HEPES, 10 glucose, adjusted to pH 7.4 with CsOH.
After cutting, we let the slices recover for 1 h at 35 °C and for another 2 h at room temperature in recording solution.
