Surgery Rats were anesthetized with urethane (1 5 g/kg, i p ) and

Surgery Rats were anesthetized with urethane (1.5 g/kg, i.p.) and placed in a stereotaxic instrument in the skull flat position and body temperature was monitored and maintained at 37°C by a thermoregulated heating pad (FHC, Bowdoin, ME). Electrode placements were mapped according to the coordinates found in the Paxinos and Watson brain atlas (Paxinos and Watson 1998) for the perforant path (7.2 mm posterior and 4.1 mm

lateral from bregma) and for dentate gyrus (3.5 mm posterior and 2.0 mm lateral). A concentric bipolar stimulating electrode (NE-100; Kopf Instruments, Tujanga, CA) was lowered into the perforant path (~3.0 mm from brain surface). A conjoined Inhibitors,research,lifescience,medical electrode/cannula assembly was constructed of a single stainless steel recording electrode (0.5–1 MΩ; FHC Inc.) and a 22-gauge stainless steel guide cannula ( Plastics One, Roanoke, VA). The cannula and electrode, secured together with regular epoxy, were aligned so that a 28 gauge injection cannula, when inserted into the guide cannula would sit, ~25 μm

lateral Inhibitors,research,lifescience,medical and 50 μm dorsal to the tip of the electrode. This ensured that the concentration delivered at the recording site was as close as possible to the concentration Inhibitors,research,lifescience,medical infused. The internal injection cannula (Plastics One) was attached to a solution-filled (ISO in aCSF or aCSF only) autoanalyzer tubing and a dH2O-filled 5 μL microsyringe. The total injection (guide and internal cannula) and recording assembly was then slowly lowered into the granule cell layer of the dentate gyrus (~2.5–3.5 mm from brain surface). The electrode placement was localized Inhibitors,research,lifescience,medical to the granule cell

layer by monitoring the response to 0.2 ms test pulses delivered to the perforant path and by maximizing the positive-going fEPSP and negative-going population spike. Stimulation and recording procedures Single monophasic square wave test pulses (0.2 ms) were delivered to the perforant path using an interstimulus interval of 30 sec (Neurodata Instruments, New York, NY). The evoked responses were amplified, filtered (0.3 Hz to 3 kHz; P5-11; Grass Instruments, West Warwick, Inhibitors,research,lifescience,medical RI), and digitized at a rate of 10 kHz and stored online for analysis. At the commencement, and at the termination of the recording period, an input–output current intensity series (I/O curve) was determined. Rutecarpine This consisted of sampling three evoked responses at interstimulus intervals (ISI) of 10 sec, at each current level from 100 to 1000 μA at 100 μA intervals. On the basis of the initial I/O curve, a current intensity for baseline current stimulation was chosen at the intensity that produced approximately 50% of the maximal population spike. DataWave software (DataWave Technologies; Loveland, CO) was used to collect waveforms and analysis was performed after the experiments (see Data Analysis and Statistics). Baseline-evoked responses were recorded every 30 sec for at least 1 h before ISO infusion began.

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