5-HT1A Autoradiography

For 5-HT_1A binding, 3.0 nM [³H]WAY-100635, 74Ci/mmol (Perkin Elmer, Waltham, MA, United States) was used. Tissue was rinsed in 50 mM Tris–HCl buffer (pH 7.5) followed by a 120 min incubation in the tracer buffer at room temperature. 10 nM of L-485,870, a dopamine antagonist, was included to prevent binding of WAY-100635 to Dopamine D4 receptors. Following the incubation period, tissue was rinsed twice in 50 mM Tris buffer at 4°C and then dipped in dH₂O and air dried. Tissue was exposed to Carestream BioMax MR Film (Kodak, Rochester, NY, United States) for 6 weeks with ³H microscale standards (American Radiolabeled Chemicals, St. Louis, MO, United States). We had no a priori predictions as far as 5-HT_1A binding sex differences at baseline for this species.