Patchmaster v2x32 program

Patch-clamp recordings were performed as previously described^{16 (link),17 (link),34 (link)}. Briefly, borosilicate glass pipettes with 4–6 MΩ resistance were used for whole-cell recordings (in mM): 122 K-gluconate, 13 KCl, 10 HEPES, 10 phosphocreatine, 4 ATP-Mg, 0.3 GTP-Na, 0.3 EGTA (adjusted to pH 7.35 with KOH). The composition of the extracellular solution was (mM): 125 NaCl, 2.5 KCl, 25 glucose, 25 NaHCO₃, 1.25 NaH₂PO₄, 2 CaCl₂, 1 MgCl₂, 10 μM pyruvic acid bubbled with 95% O₂ and 5% CO₂. Signals were amplified using an EPC10-2 amplifier (HEKA Elektronik, Lambrecht, Germany). All recordings were performed at 34 °C using a temperature control system (Bath-controller V, Luigs & Neumann, Ratingen, Germany) and slices were continuously superfused at 2–3 ml/min with the extracellular solution. Current- and voltage-clamp recordings were sampled at 10 kHz, with the Patchmaster v2x32 program (HEKA Elektronik).

Patch-clamp recordings were performed as previously described (Fino et al., 2010 (link); Paillé et al., 2013 (link); Cui
et al., 2015). For whole-cell recordings borosilicate glass pipettes of
4-6 MΩ resistance contained (in mM): 105 K-gluconate, 30 KCl, 10 HEPES, 10
phosphocreatine, 4 ATP-Mg, 0.3 GTP-Na, 0.3 EGTA (adjusted to pH 7.35 with KOH).
The composition of the extracellular solution was (mM): 125 NaCl, 2.5 KCl, 25
glucose, 25 NaHCO₃, 1.25 NaH₂PO₄, 2
CaCl₂, 1 MgCl₂, 10 μM pyruvic acid bubbled with 95%
O₂ and 5% CO₂. Signals were amplified using EPC10-2
amplifiers (HEKA Elektronik, Lambrecht, Germany). All recordings were performed at
34°C using a temperature control system (Bath-controller V, Luigs&Neumann,
Ratingen, Germany) and slices were continuously superfused at 2–3 ml/min with the
extracellular solution. Slices were visualized on an Olympus BX51WI microscope
(Olympus, Rungis, France) using a 4x/0.13 objective for the placement of the
stimulating electrode and a 40x/0.80 water-immersion objective for localizing
cells for whole-cell recordings. Series resistance was not compensated.
Current-clamp recordings were filtered at 2.5 kHz and sampled at 5 kHz and
voltage-clamp recordings were filtered at 5 kHz and sampled at 10 kHz using the
Patchmaster v2x32 program (HEKA Elektronik).