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Betamethasone 0.025 mg/kg of saline for 4 days and then daily dosing with saline at 0.025 mg/kg. These doses were chosen to achieve a minimum of 5% the maximum attainable weight (maximum effective dose). The animals were tested for locomotion behavior with three different experimental groups (n = 6): 1) Vehicle-injected (veh/saline) controls; 2) Vehicle-injected (veh/saline) rats receiving saline for 4 days, and 3) Vehicle-injected (Veh/saline) rats receiving saline for 4 days, but not and then receiving treatment with either vehicle (veh/saline) or vehicle/saline (Veh/saline/saline). Treatment with either vehicle or vehicle/saline was used in order to assess locomotion. The locomotor activity was determined visually using an infrared tracking device as described above. The frequency of movements Cheap zopiclone for sale in uk in presence a new stimulus was calculated and displayed on a computer screen. The animals were sacrificed by cervical dislocation (25-gauge needle) and perfused transcardially with 4% paraformaldehyde (PFA). Brains from these animals were removed on ice and placed in 10% (w/v) phosphate-buffered neutral formalin for 30 minutes at room temperature. Brains were removed from the hippocampus and snap-frozen in liquid nitrogen stored at -80°C. The striatum was excised in a 10-μm-thick coronal section and prepared for immunohistochemical staining using the method described by Faucher et al. (19). Briefly, the sections were blocked with goat serum (2% normal in phosphate-buffered saline, pH 7.4, 150 μls/ml), followed by incubation in mouse anti-α7 nAChR (1:10; Abcam, Cambridge, MA), as previously described (18). After the sections were washed with PBS, the sections were incubated with a primary antibody conjugated to secondary horseradish peroxidase-conjugated antibody (Jackson Immuno Research, West Grove, PA), for 1 hour at room temperature. Following washing with PBS, the sections were developed with a standard chemiluminescence kit (Jackson Immuno Research, West Grove, PA). The images were Where to buy zolpidem online captured using a Zeiss LSM 510 confocal microscope and analyzed using a Leica DM6000 fluorescence microscope (Leica Microsystems GmbH). Immunohistochemical analysis The striatum was dissected and stained with the primary anti-α7 nAChR (1:10,000; Abcam, Cambridge, MA; 1:800,000, 1:1000). The tissue was then immersed in 0.05% Triton X-100 PBS for 1 hour. The tissue was then washed and mounted with a mounting medium (0.05% Triton X-100 in PBS). The striatal sections were then photographed using a fluorescent microscope (Leica DM6000). Images were analyzed on a Leica DM6300 digital camera. Behavioral evaluation of the mice Each mouse was tested in a novel experimental apparatus order to evaluate the effects of two different anti-nAChR agonists on the mice's behavior. experimental apparatus consisted of an open-field (12 × 12 20 cm) filled with white PFA, a light-dark box (7 × 7 5 cm) and a light-dark/dark box (7 × 7 5 cm). The mice were meridia pillen kaufen tested individually or, if necessary, in groups of three. All experiments were conducted in accordance with the approved guidelines of Animal Care and Use Committee of the University Zurich. The mice were tested in open-field apparatus as previously described (16). The latency to enter light-dark box was evaluated as previously described (16, 17). Behavioral analysis of the mice The latency of mice to enter the light-dark box was evaluated with (7 × 7 5 cm). The latency of mice to enter the light-dark box was tested in a separate session. The latency was measured by placing a light light-dark box around the cage and measuring number of seconds the mouse remained in light box. The average number of seconds mouse remained in the light-dark box, was calculated by determining the average number of seconds mouse remained in the light-dark box during 10-s period in which the light was turned on and off. The average number of seconds mouse remained in the light-dark box was expressed as a percentage of total number seconds that the mouse remained in light-dark box.

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