3–5,000 Hz) and unit activities (300–5,000 Hz). Raw ECoG signal was band-pass filtered (0.3–1,500 Hz) and amplified (2,000×). All signals were digitized online at 16.67 kHz using a Power 1401 analog-digital converter (Cambridge Electronic Design) and stored on a PC running Spike2 software (versions 6.08 and 6.09, Cambridge Electronic Design). GABAergic cell recordings lasted 15–105 min (typically ∼45 min). The juxtacellular recording mode (rather than, for example, a quasi-intracellular mode), was assured by only including for analysis neurons that (1) had

stable spontaneous firing rates/patterns and stable spike widths; (2) did not display any “injury discharge”; and (3) were recorded in the absence of spurious “baseline noise” or hyperpolarizing Ipatasertib mw shifts in the electrode potential. After recordings, neurons were selectively filled with Neurobiotin using Selleckchem MEK inhibitor juxtacellular labeling (Pinault, 1996).

Spike shape and amplitude were monitored throughout recording and labeling to ensure that the same neuron was recorded and labeled. In order to verify the location of the reference electrode, an extracellular Neurobiotin deposit was made in the dorsal CA1 (100 nA anodal current 1 s, 50% duty cycle for 20–30 min). Only data acquired before labeling and obtained from unequivocally identified cells were analyzed. All data were analyzed off-line using Spike2 built-in functions and custom scripts (Tukker et al., 2007). Spikes were detected with an amplitude threshold in the BLA unit channel. Occasionally, crotamiton additional smaller amplitude units were present in the recording. Spike2 clustering function supervised manually was used to isolate single units, and identity of labeled neurons was systematically ensured as described above. Spike sorting was always checked using autocorrelograms, which showed clear refractory periods (≥2 ms). Hippocampal theta oscillation epochs

were detected by calculating the theta (3–6 Hz) to delta (2–3 Hz) power ratio in 2 s windows of the dCA1 LFP (Csicsvari et al., 1999 and Klausberger et al., 2003). Ratio >4 in at least three consecutive windows marked theta episodes. We excluded from this analysis periods of noxious stimuli and the following 20 s. Every theta episode was visually checked. Selected periods always consisted of robust theta oscillations. They exclusively occurred during persistently activated brain state (Figure S9). After theta episodes detection, the dCA1 LFP was downsampled to 1.04 kHz, digitally filtered (3–6 Hz) and the troughs were determined (Spike2). Each spike was assigned an angle relative to surrounding theta troughs (Tukker et al., 2007 and Klausberger et al., 2003). The precision of our electrode placements (mediolateral and antero-posterior ranges ∼400 μm) ensured phase consistency between experiments (i.e., ∼8.5 degrees error, assuming a phase shift of 21°/mm; Lubenov and Siapas, 2009).