Rabbit anti zinc transporter 3 znt3

Slide mounted brain sections (two per slide) from dorsal hippocampus were processed for histological studies. Sections were immunostained with the following antibodies: rabbit anti-pS6 (1:500, Cell Signaling), chicken anti-GFP (1:500, Abcam, Boston, MA), rabbit anti-zinc transporter 3 (ZnT3) (1:3000, Synaptic Systems, Gottingen, Germany), rabbit anti-Glutamate receptor 2/3 (1:100, Millipore, Temecula, CA), rabbit anti-prox1 (1:1000, Sigma-Aldrich, St. Louis, MO), rabbit anti-GFAP (1:500, Millipore) and rabbit anti-Iba1 (1:1000, Synaptic Systems). AlexaFluor488 goat anti-chicken, AlexaFluor488 goat anti-rabbit, AlexaFluor594 goat anti-rabbit and AlexaFluor647 goat anti-rabbit secondary antibodies were used (Invitrogen, Eugene, OR). Sections were dehydrated in alcohols, cleared in xylenes and coverslips were attached with Krystalon mounting-media (Harleco, Darmstadt, Germany).

At 1, 2, 4 or 8-10 weeks post SE or sham treatment, rats were deeply anesthetized with pentobarbital and transcardially perfused with 0.9% saline and 4% paraformaldehyde (PFA). Brains were removed and post-fixed overnight in 4% PFA. Sixty μm sections were cut in the coronal plane using a vibrating blade microtome (VT1000S, Leica Microsystems Inc, Buffalo Grove, IL). Representative sections (4-6 sections, 720 μm apart) through the rostro-caudal axis of the brain were processed with standard fluorescent immunohistochemical techniques using the following primary and secondary antibodies: mouse anti-bassoon (1:1000, Enzo Life Sciences), rabbit or chicken anti-GFP (1:2000 for both, Invitrogen and Aves), mouse anti-striatal enriched phosphatase (STEP) (1:1000, Cell Signalling), rabbit anti-zinc transporter-3 (Znt3) (1:3000, Synaptic Systems), or mouse anti-regulator of G-protein signaling 14 (RGS14) (1:100, NeuroMab). Fluorescent signals were achieved using Alexa Fluor secondary antibodies: goat anti-mouse 594, goat anti-mouse 647, goat anti-rabbit 488 or 594, and goat anti-chicken 488.