Subjects were anesthetized with sodium pentobarbital (0.1 mg/10 g body weight) and perfused through the ascending aorta with 0.9% saline, followed by 4% paraformaldehyde in 0.1 M phosphate buffer solution (PBS; pH 7.4). Brains were harvested, postfixed for 2 hrs in 4% paraformaldehyde, and then stored in 30% sucrose in PBS. All brains were cut into 30–40 μm coronal sections on a microtome, and the floating sections were stored in 0.1 M PBS with 0.01% sodium azide at 4°C until processing for immunocytochemistry.
