The primary antibodies used in this study were rabbit polyclonal antibodies to calcyon (created and affinity purified in-house; described by Lezcano et al26), mouse monoclonal antibodies MAB1680 to filamin-A (Chemicon International, Temecula, Calif) (described by Aakhus et al37), and rabbit polyclonal antibodies RU-144 to spinophilin (described by Allen et al38 and Lidow et al39). To evaluate the specificity of these antibodies, we examined their labeling of Western blots of the dorsolateral prefrontal cortical tissue from 5 monkey and 3 randomly selected human cases used in this study. For this purpose, the tissue homogenates (3 µg per well) were loaded into ready-made 4% to 15% gradient sodium dodecyl sulfate gels (Bio-Rad Laboratories, Hercules, Calif) and run for 1.5 hours at 100 V using a Ready Gel Cell with Tris/glycine/sodium dodecyl sulfate running buffer (Bio-Rad Laboratories). Separated proteins were transferred to polyvinylidene difluoride membranes (Osmonics, Minnetonka, Minn) for 2 hours at 100 V also using a Ready Gel Cell with Tris/glycine buffer (Bio-Rad Laboratories). Before immunostaining, membranes were preincubated for 1 hour at room temperature in blocking solution consisting of 0.1M phosphate buffer (pH 7.6), 5% fat-free dry milk, and 0.1% Tween 20. Incubation with the primary antibodies for calcyon(8 µg/mL), filamin-A (5 µg/mL), or spinophilin (5 µg/mL) in the same blocking solution was conducted overnight at 4°C. The membranes were next exposed for 1.5 hours at room temperature to horseradish peroxidase–conjugated goat anti–rabbit or anti–mouse secondary antibodies (Sigma-Aldrich Corp, St Louis, Mo) (1 µg/mL) in the blocking solution described herein. Visualization of the labeling was conducted using a chemiluminescence substrate (Super Signal; Pierce Biotechnology, Rockford, Ill). The images were produced by opposing transparent plastic-wrapped chemiluminescence-soaked membranes to an autoradiography film (X-Omat AR; Eastman Kodak Co, Rochester, NY) for 1 to 45 minutes. In addition, antibody specificity was tested by preabsorption with peptides (100 µg/mL) against which the antibodies were raised. The preabsorbed antibodies were applied to slot blots of prefrontal cortical tissue from 5 monkeys and 3 randomly selected human cases in a manner described in the following subsections.