The ECGs were measured for a cumulative total of 40 s of recording in 1-s samples. Half of the 40 data segments were when the monkeys were ‘asleep’ and half whilst they were ‘awake’. The recorded potentials were sampled at 100 Hz and PI3K activity low-pass filtered to include the frequency range 0–50 Hz. The power spectra of the ECG were then calculated separately for

awake (BS3) and sleep states (BS1) using the spectral calculation performed by fast Fourier transform (FFT) methods, utilizing the procedures and C code described by Press et al. (1992). The use of multiple independent data segments to compute an average of the power spectra for each state ensured that the resulting power spectra for each state were statistically reliable, as described elsewhere (Press et al., 1992; Bendat & Piersol, 2010). The ECGs demonstrated that when the subjects were rated by the experimenter as being in BS3 (eyes-open/awake) the ECG showed low-voltage fast activity, and this was reflected in the power spectra (range 2–20 Hz) which had a peak in the frequency range 23–28 Hz, as shown in Fig. 2. Increased power at low frequencies

is a sign of SWS (Finelli et al., 2001). When the subjects were rated by the experimenter as being in BS1 (eyes-closed/asleep), high-voltage slow waves appeared in the ECG, and this was reflected in the power spectra with relatively more power than when awake in the lower frequencies between 5 and 18 Hz (which include the alpha and theta bands), as illustrated in Fig. 2. The power spectra shown in Fig. 2, taken Selleck RAD001 together with similar data obtained in other macaques (Rolls et al., 2003), confirm the experimenter’s assessment of the behavioural states as BS3 or ‘awake’ (i.e. periods when the monkeys had their ‘eyes-open’), and as BS1 or ‘asleep’ (i.e. when the animals had Histone demethylase their ‘eyes-closed’). Cells in mPFC showing responses to eye-closure or eye-opening could be classified on the basis of their firing rate changes during transitions between behavioural states (see Figs 3-7). Type 1 cells significantly

increased their firing rate when the subjects closed their eyes and went to sleep, and returned to their previous levels on reopening of the eyes. Type 2 cells significantly decreased their firing rate on eye-closure, and returned to their former level of activity with eye-reopening. Type 3 cells were unaffected by both eye-opening and eye-closure. Neuron firing rates were recorded every 10 s as described above for periods of many minutes that could include several (up to nine) discrete periods of eye-closure/eye-opening (Fig. 4). Mean firing rates were calculated separately for each BS3, BS2 and BS1 epoch. Mean epoch values were then used to obtain the overall mean BS3, BS2 and BS1 firing rates for each neuron. ‘Grand mean’ firing rate estimates (together with standard error values) for each behavioural state (BS1, 2 and 3) were subsequently generated for each of the three cell types 1–3 (Table 1).