Spike sorting was performed by a semiautomatic clustering procedu

Spike sorting was performed by a semiautomatic clustering procedure (see Supplemental Experimental Procedures and Figure S5), yielding a total of 140 active pyramidal neurons during sleep (n = 4

animals), 573 in the maze (n = 3 animals), 569 during wheel running (n = 3 animals) and 193 in the open field (n = 4 animals). The determination of place fields was basically performed as in Leutgeb et al. (2007). For each cell, spatial distributions of firing rates (ratio of total number of spikes to occupancy duration in a given spatial bin) during locomotion (>5 cm/s) were calculated for each bin of the environment (50 × 50 bins) and then boxcar-averaged over the 25 neighboring bins (instead of a Gaussian kernel). Place fields included the bins with the highest firing rates (at least 2 Hz) and all contiguous bins in which the firing rate exceeded 20% buy Ulixertinib of the peak firing rate. Place fields smaller selleck than 16 bins or larger

than half the environment were discarded. Episode fields (EpF) in the wheel were defined as periods <3 s with peak firing rate >5 Hz and 4SD above mean firing rate, EpF limits were set at 10% of the peak firing rate, as described in Pastalkova et al. (2008). Because theta is reliably expressed when the animal is moving, we have selected for analysis all awake periods with running speed >10 cm/s (although including brief interruptions of less than 200 ms). REM sleep periods were determined from manual threshold on theta/delta ratio, which provides a well contrasted estimation of REM sleep versus slow-wave state (Figure S6). Periods shorter than 3.5 s were discarded. Time-frequency spectrograms of continuous LFP traces (EEG, sampling rate 1.25 kHz, hardware Resminostat high-pass filter 1 Hz) were computed by using the multitaper method (1 s time window and

4 tapers) from the Chronux toolbox (Mitra and Bokil, 2008; http://chronux.org/). Theta power signal was measured from the raw EEG trace as the integrated power in the 4–11 Hz frequency band by using the multitaper method. For second-order analysis, theta power was also measured as the peak-to-trough amplitude of individual theta cycles in the 2–30 Hz filtered (“raw theta”) EEG trace or by using Morlet-based wavelet analysis (Bruns, 2004). These methods yielded similar results (Figures 2, S2, and S6). Local maxima (peaks) and minima (troughs) of theta power were detected and measured on the theta power trace, within 200 ms of the peaks and troughs obtained from the low-pass filtered theta power signal (fourth-order Chebychev, cutoff 1.8 Hz; Figure S7). TPSM cycles were defined as the intervals between successive theta power minima and a linear phase (from 0 to 2π) was defined between the successive troughs of TPSM cycles (see Supplemental Experimental Procedures).

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