utexas.edu/djlab). The apparatus was consisted of opaque- and matte-finished black acrylic sheet (36” × 36” × 24”).

Each rat was randomly assigned and placed into the center of the field following 30 min saline, vehicle, and diazepam i.p. injection. Behavior in the open field arena was recorded for 6 min using a CCD camera. The surface of the open field arena was cleaned with 70% EtOH in order to remove permeated odors by previous animals after each trial. To analyze behaviors Selleck JAK inhibitor for the last five minutes, the open field arena was divided into 9 equal-size squares (12” by 12”). Basal exploration activity was measured by total traveled distance (inch) and anxiety level was assessed by the number of center square entries, the duration, Vorinostat nmr and the traveled distance in the center square. A line crossing was defined as the body center crosses a line. The apparatus was made of black acrylic sheet. Four arms (50 cm long and 10 cm wide) are connected and elevated to a height of 50 cm from the floor. Two

arms are open (open arms) and the other two arms are enclosed within 40 cm walls. Each rat was placed into the intersection of the four arms (center area) 30 min following saline, vehicle, or diazepam i.p. injection. Behavior in elevated plus maze was recorded for 6 min using a CCD camera. The surface of the elevated plus maze was cleaned with 70% EtOH in order to remove permeated click here odors by previous animals after each trial. To

analyze behavior test for 6 min, the number of arm entries and the percentage of open arm time (duration in open arm/total time) were assessed. Arm entry was defined as 50% of the body being positioned within the arm. The tank is made of transparent Plexiglas cylinder (80 cm tall × 30 cm in diameter) filled with water (23°C–24°C) to a depth of 45 cm. Water in the tank was changed after each trial. For the first exposure, rats without drug treatment were placed in the water for 15 min (pretest session). Twenty-four hours later, rats were placed in the water again for a 6 min session (test session) 30 min following saline, vehicle, diazepam (1 mg/kg), ketamine (15 mg/kg), or fluoxetine (10 mg/kg) i.p. injection. Forced swim test was recorded by a video cameras positioned on the top of the water tank. A passive activity was defined as floating and making only those movements necessary to keep the nose above the water. Behaviors from forced swim test was quantified by using a time sampling technique to rate the predominant behavior over a 5 s interval as described (Lucki, 1997) and a custom written program. Four- to five-week-old rats were bilaterally microinjected with lentivirus expressing shRNA-control or shRNA-HCN1 in the dorsal hippocampal CA1 region. After behavior test, dorsal hippocampal slices (350 μm) were prepared from 10- to 12-week-old lentivirus-infected male Sprague-Dawley rats.