Recordings were from either sex, as previously described (17 (link), 53 (link)). Slices were perfused with a flow rate of 2 ml/min at 30°C, with ACSF equilibrated with 95% O2 and 5% CO2 (final pH 7.3). Schaffer collaterals were stimulated every 15 s with a bipolar tungsten electrode, and resulting fEPSPs in CA1 were recorded with a glass electrode filled with ACSF. Signals were amplified with an Axopatch 2B amplifier, digitized with Digidata 1320A, and analyzed with Clampex 9 (Molecular Devices). Stimulus strength was titrated to define maximal response and input-output curves and adjusted to result in ~50% of maximal response. PTT-LTP was induced by a 3-min, 5-Hz tetanus. The average of fEPSP initial slopes from the 5 min preceding the tetanus was set to equal 100% baseline level. The PTT-LTP strength was defined as the average of fEPSP initial slopes obtained between 15 and 45 min after the tetanus. To determine paired-pulse facilitation, fEPSP initial slopes of two consecutive stimuli with the indicated interevent intervals were recorded.
Axopatch 2b amplifier
Manufactured by Molecular Devices
The Axopatch 2B is a high-performance patch-clamp amplifier designed for electrophysiology applications. It provides accurate measurements of ionic currents across cell membranes, enabling the study of ion channel activity and cellular function.
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2 protocols using axopatch 2b amplifier
1
Schaffer Collateral Synaptic Plasticity
2
Endogenous Chloride Determination in Neurons
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Neurons were recorded at 33 °C in saline containing (in mM) 140 NaCl, 2.5 KCl, 2.5 MgCl2, 2.5 CaCl2, 10 HEPES, 11 glucose, pH 7.4 (NaOH). All solutions were applied through a three-barrel microperfusion apparatus (700 μm, Warner Instruments, Hampden, CT), and conducted in the presence of tetrodotoxin (300 nM) and bumetanide (10 μM) to block voltage-dependent sodium channels and NKCC1, respectively. Cells infected with control- or Cre-containing virus were selected by epifluorescence for GFP expression. Data were acquired using Clampex 10 software at an acquisition rate of 10 kHz with an Axopatch 2B amplifier and digitized with a Digidata 1440 (Molecular Devices). To detect endogenous Cl− levels, cells were perforated with gramicidin (50 mg/mL) dissolved in an internal pipette containing (in mM) 140 KCl, 10 HEPES, pH 7.4 (KOH) and a tip resistance of 3–4 MΩ. Following perforation to below a series resistance of 100 MΩ, EGABA values were found by application of muscimol (1 μM) during positive-going voltage ramps (20 mV/s).
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