Male and female mice age P30-P50 were euthanized with isoflurane, brains removed and horizontal slices prepared from midbrain (200µm) in ice-cold artificial cerebral spinal fluid containing (in mM): NaCl 119, KCl 2.5, MgCl2 1.3, CaCl2 2.5, NaH2PO4 1.0, NaHCO3 26.2 and glucose 11, pH 7.2, bubbled with 95% O2 and 5% CO2. Slices were equilibrated for ~45 min at 33° C in ACSF supplemented with (in mM): L-Ascorbic acid 0.4, Na Pyruvate 2, myo-Inositol 3, then transferred to a recording chamber equipped with constant perfusion of ACSF (2 ml/min).
