Multiclamp 700b amplified

Whole-cell recordings in CA3 pyramidal neurons were performed using 3-3.5 MΩ pipettes with an intracellular solution (in mM): K Gluconate 130, KCl 5, MgCl₂ 2, HEPES 10, di-tris-Phosphocreatine 10, NaATP 4, NaGTP 0.4 (pH adjusted to 7.2 with KOH, osmolarity 290-295 mOsM). The morphological tracer Alexa Flour 594 hydrazide (50μM) was also included in the intracellular solution. Voltage clamp recordings of miniature EPSCs were made, with the mEPSCs recorded at -70 mV. Receptor agonists and antagonists purchased from Tocris Cookson (Bristol, UK) were added to the aCSF at the following concentrations (in μM): TTX 1, Picrotoxin 100 and DCG-1V ((2S, 2′R, 3′R)-2-(2′, 3′-dicarboxycyclopropyl) glycine) 2. Recordings were performed with a Multiclamp 700B amplified (Molecular Devices). Recording sweeps were collected at 5kHz using WinWCP (Strathclyde Electrophysiology). mEPSCs were recorded over 15min periods in control and treated conditions, data were then analyzed off-line using Mini-Analysis software (Synaptosoft). mEPSCs were detected if their amplitude was greater than the threshold of 5pA, and considered for analysis if their rise time was shorter than their decay time. Fluorescentprobes were purchased from Invitrogen.