Zebra Finch Brain Slice Preparation

Adult male zebra finches (Taeniopygia guttata; >120 days) were obtained from a supplier. The animal studies were approved by the Institutional Care and Use Committee of Jiangxi Science and Technology Normal University. The experimental methods were described in our previous studies (Wang et al., 2014 (link), 2015 (link); Meng et al., 2016 (link)). In brief, birds were anesthetized and euthanized by decapitation. The brains were dissected and immersed in ice cold, oxygenated (5% CO₂ and 95% O₂) solution containing (in mM) 62.5 NaCl, 5 KCl, 28 NaHCO₃, 10 glucose, 1.3 MgSO₄⋅7H₂O, 1.26 NaH₂PO₄⋅H₂O, and 248 sucrose (pH 7.4). Coronal brain slices measuring 250–300 μm in thickness and containing RA were obtained using a vibrating microtome (NVSLM1, World Precision Instruments, USA). Slices were collected at 37°C in oxygenated artificial cerebrospinal fluid (ACSF) containing (in mM) 25 glucose, 25 NaHCO₃, 1.27 NaH₂PO₄⋅H₂O, 2.5 KCl, 1.2 MgSO₄⋅7H₂O, 2.0 CaCl₂, and 125 NaCl (pH 7.4). After 30 min, the slices in the holding chamber were allowed to recover at room temperature (22–26°C) for 1 h.