Vibratome 1500

Animals were anesthetized with ketamine (100 mg/kg) and xylazine (10 mg/kg) in saline. For protein isolation (animals aged P7-P45 for Figure 1A and P28 for Figures 1F and 1G), visual cortices were isolated according to stereotactic coordinates (0.5–1 mm anterior to λ, 2–3 mm lateral to midline) followed by sonication in 8 M urea. For LGN isolation, forebrain was flash frozen over liquid nitrogen and later dissected on an iced platform. LGN was visually identified and isolated with a tapered scalpel blade. For GABA and glutamate receptor immunoblots, crude synaptoneurosomes were prepared as described (Villasana et al., 2006 (link)). Protein concentrations were determined using the BCA method (Thermo-Fisher Scientific, Holtsville, NY). For microscopy, animals (P7-P70, as indicated in figure legends) were transcardially perfused first with ice cold PBS and then with 4% PFA (in PBS, pH 7.4). Brains were isolated and postfixed overnight in 4% PFA and washed overnight in PBS (all at 4°C). Brains were then embedded in 3% agarose in PBS and sectioned at 40–60 μm using vibrating microtome (Leica VT1000, Leica Biosystems, Nussloch, Germany or Vibratome 1500, Harvard Apparatus, Holliston, MA). Sections were stored in PBS with 0.01% sodium azide (Sigma-Aldrich, St. Louis, MO) at 4°C.