The rats were administered ketofen (3–5 mg/kg) to minimize pain and returned to normal housing to recover

for 3–5 days. OPTICAL STIMULATION AND ELECTROPHYSIOLOGIC RECORDINGS Using our adapted NeuroRighter system, electrophysiologic recordings were sampled at 25 kHz with a 1–9,000 Hz bandwidth. LFPs were isolated online with a 1–500 Hz 1-pole Butterworth band-pass PLK inhibition selleck filter and downsampled to 2000 Hz. Action potentials were isolated both online (Newman et al., 2013) and offline, with the offline results presented here. Action potentials were detected offline using custom-written adaptations to the automated spike-sorting Wave_clus scripts (Quiroga et al., 2004). The raw data was band-pass filtered offline from 500 to 5000 Hz. For the TDT electrodes, the median signal was removed across the CA3 and CA1 electrodes, respectively. For the NeuroNexus Array, the median signal was removed across all electrodes. Positive and negative thresholds were applied at 5x the SD of the signal, and the resulting waveforms were matched, sorted, and isolated using

superparamagnetic clustering (Wave_clus; Quiroga et al., 2004). Power spectra and spectrograms were computed using the Chronux suite of analysis tools and multitaper analysis (Bokil et al., 2010), with a moving window size of 4 s stepping in 0.5 s increments, T = 4, W = 1, and seven tapers. Data were recorded intraoperatively and for up to 4 weeks postoperatively. To stimulate awake and behaving animals, calibrated ferrules were connected via armored patch fiber cables (200 μm diameter, 0.67 NA, Plexon). Square-wave stimulation pulses varied between 10, 30, and 50 mW/mm2; 7, 11 (theta), 17, 23, 35 (beta), and 42 (gamma) Hz; and 2, 5, and 10 ms pulse widths. NeuroRighter enables custom-designed

stimulation times and amplitudes to be defined via Matlab script (Newman et al., 2013). We leveraged this customizability to develop several other stimulation patterns, including varying frequency, Poisson distributions, and continuous sinusoids, which are described in more detail as they are presented. In all cases, the experimental protocol consisted of repeated 1 min recordings of 20 s of background, 20 s of stimulation with a particular pattern, and a subsequent 20 s of additional background. Stimulation protocols were performed AV-951 in random order and repeated numerous times over several recording sessions. This setup was able to stimulate and record LFP and single-unit responses from awake and behaving animals uninterrupted for several hours and over several days. HISTOLOGY Histology was performed after experimentation to verify microelectrode recording locations and light-sensitive ion channel expression. Rats were deeply anesthetized with an overdose of Euthasol (5 ml/kg, Virbac, Fort Worth, TX, USA) injected intraperitoneally. They were then transcardially perfused with 0.9% saline followed by 4% paraformaldehyde in 0.1 M phosphate buffer at pH 7.2.