Tris gtp

Brain slices were transferred to a recording chamber constantly perfused with 32 to 34°C warm ACSF containing (in mM): 119 NaCl (Sigma-Aldrich), 2.5 KCl, 1 NaH₂PO₄, 1.3 MgCl₂, 26 NaHCO₃, 10 D (+)-glucose and 2.5 CaCl₂ (all from Merck, Darmstadt, Germany) bubbled with a gas mixture of 95% O₂ and 5% CO₂. Cortical pyramidal neurons were visualized in layer 5 with an Axioskop 2 FS + microscope (Carl Zeiss MicroImaging GmbH, Göttingen, Germany) equipped with infrared differential interference contrast. Patch pipettes were pulled (P-97 micropipette puller, Sutter Instruments, Novato, CA, USA) to a resistance of 3 to 5 MΩ. For recordings with 4β-PMA and Bryostatin1 intracellular solution comprised (in mM) 120 K-gluconate, 10 Na-phosphocreatine, 11 EGTA, 2 Mg²⁺ATP, 0.3 Tris-GTP (Sigma-Aldrich), 10 KCl, 1 MgCl₂, 1 CaCl₂ and 10 HEPES. For experiments with IFN-β, pipette solution contained (in mM): 120 K-methylsulphate (KMeSO₄) (ICN Biomedical Inc, California, USA), 20 KCl, 14 Na-phosphocreatine, 4 NaCl, 0.5 EGTA, 10 HEPES, 4 Mg²⁺-ATP, 0.3 Tris-GTP and 0.1 cAMP (Sigma-Aldrich). The pH of intracellular solutions was adjusted with KOH (Carl Roth, Karlsruhe, Germany) to 7.2.

cesium-methanosulfonate (Sigma-Aldrich, cat# C1426), Na₂-ATP (Sigma-Aldrich, cat# A26209), Tris-GTP (Sigma-Aldrich, cat# G9002), Tris-phosphocreatine (Sigma-Aldrich, cat# P1937), EGTA (Sigma-Aldrich, cat# 324626), CsOH (Sigma-Aldrich, cat# 562505), Na-ascorbate (Sigma-Aldrich, cat# A4034)