Whole-cell patch clamp recording was performed as reported previously [82, 83]. Mice were anesthetized with isoflurane and coronal slices (300 μm) of spinal cord were prepared. After 1 h of recovery, brain slices were perfused with oxygenated artificial cerebrospinal fluid (ACSF) solution at 3–4 ml/min. NaCl was replaced with NaMeSO3 to eliminate potential Cl- currents. Whole-cell patch-clamp recordings in microglia were made using 5–10 MΩ glass pipettes filled with a TMA-based intracellular solution consisting of 100 mM TMA-MeSO3, 1 mM EGTA, and 100 mM MES (pH 5.5, 290–300 mOsm). The membrane potential was held at − 60 mV. Data were amplified and filtered at 2 kHz by a patch-clamp amplifier (Multiclamp 700B), digitalized (DIGIDATA 1440A), stored, and analyzed by pCLAMP (Molecular Devices, Union City, CA). Data were discarded when the input resistance changed > 20% during recording. A minimum of five cells from at least three different mice from the same litter were randomly selected for recording per condition.

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