Isoflurane

CAV2-Cre (0.5 μl, 3 × 10¹² viral genomes per μl) was stereotaxically injected bilaterally into the dorsal striatum (0.5 mm anterior to Bregma, ± 1.75 mm lateral to midline, 3.0 mm ventral from the skull surface) using the Allen Mouse Brain Atlas as a guide.^{41 (link)} All animals used for behavioral experiments received striatal CAV2-Cre injection: CAV2-Cre injected Slc17a6^+/+ animals were used as controls (CAV2^Cre-Slc17a6^+/+ controls) and CAV2-Cre injected Slc17a6^lox/lox littermates were used to generate CAV2^Cre-Slc17a6^lox/lox mice.
For electrophysiological experiments, CAV2^Cre-Slc17a6^+/+ controls and CAV2^Cre-Slc17a6^lox/lox littermates also received stereotaxic injections of the opsin AAV2-EF1α-DIO-hChR2(H134R)-mCherry (0.5 μl AAV2-ChR2; 3 × 10⁹ viral genomes per μl; UNC Vector Core) into the CL/CM/PF thalamic nuclei (−2.18 mm anterior to Bregma, ± 0.5 mm lateral to midline, 3.3 mm ventral from the skull surface).
During stereotaxic injections, animals were anesthetized with 1–2% isoflurane (Kent Scientific, Torrington, CT) and with lidocaine/bupivacaine (1.0 mg/kg and 0.5 mg/kg s.c.). After surgeries, analgesia was supplied by ketoprofen (5.0 mg/kg, s.c.). All animals were allowed to recover from surgeries for 4 weeks before they were submitted to further experimental procedures.

Gas anesthetics desflurane, isoflurane, and sevoflurane were bought from Kent Scientific, USA. Paclitaxel and 5-fluorouracil were bought from Sigma, USA.

Rats were deeply anesthetized with isoflurane (Kent Scientific) and euthanized by decapitation. The brain was rapidly dissected and glued on a platform submerged in an ice-cold oxygenated (95% O₂/5% CO₂) cutting solution containing (in mM): 206 sucrose (Sigma-Aldrich), 10 D-glucose (Sigma-Aldrich), 1.25 NaH₂PO₄ (Sigma-Aldrich), 26 NaHCO₃ (Sigma-Aldrich), 2 KCl (Fisher Chemical), 0.4 sodium ascorbic acid (Sigma-Aldrich), 2 MgSO₄ (Sigma-Aldrich), 1 CaCl₂ (Sigma-Aldrich), and 1 MgCl₂ (Sigma-Aldrich). A mid-sagittal cut was made to divide the two hemispheres, and coronal brain slices (300 μm) were cut using a vibrating blade microtome (Leica VT1200). The brain slices were transferred to a holding chamber with oxygenated artificial CSF (ACSF) containing (in mM): 119 NaCl (Sigma-Aldrich), 2.5 KCl (Fisher Chemical), 1 NaH₂PO₄ (Sigma-Aldrich), 26.2 NaHCO₃ (Sigma-Aldrich), 11 D-glucose (Sigma-Aldrich), 1 sodium ascorbic acid (Sigma-Aldrich), 1.3 MgSO₄ (Sigma-Aldrich), and 2.5 CaCl₂ (Sigma-Aldrich; ∼295 mOsm, pH 7.2–7.3) at 37°C for 20 min and then room temperature for at least 40 min of rest. The slices were kept submerged in oxygenated ACSF in a holding chamber at room temperature for up to 7–8 h after slicing.