En to a computer-assisted data acquisition system CED 1401 data processor (CED, Cambridge, UK). After the habituation period, polygraphic recordings of body temperature, spontaneous activity, EEG, and EMG were collected continuously, throughout the L/D cycle. Telemetric signals of body temperature were sampled by Dataquest ART (Data Sciences Int., USA). Spontaneous activity was also recorded via signal strength of the telemetric device for body temperature. EEG and EMG signals were sampled by the Spike2 acquisition program (Cambridge Electronic Design, UK). Offline sleep scoring was first done via Spike2 software, and carefully verified by visual assessment of the EEG and EMG activities. Vigilance states were classified over 6-s epochs as wakefulness, NREM, or rapid eye movement (REM) sleep. NREM sleep was characterized by a continuous, slow, high-voltage EEG and low-voltage EMG activity. REM sleep was characterized by low-voltage EEG with continuous theta waves and total suppression of the EMG. Fast Fourier Transform using the Spike2 analysis program (Cambridge Electronic Design, UK) calculated the EEG power spectrum in the epoch determined to be NREM sleep. The sampling rate for EEG/EMG data collection was 100 Hz. The EEG delta and theta frequency band was set at 0.5?.0 Hz and 4.0?.0 Hz, respectively. Each value of power 16985061 was presented as a percentage of the total power (0.5?0 Hz). In this study, we used delta/theta value as a parameter of SWA [26]. Body temperature, spontaneous activity, sleep/wake durations, and EEG delta/theta ratio were averaged for hourly intervals. After the 24-hour baseline-recording period, Title Loaded From File 6-hour SD by gentle touching with a brush was carried out, starting at 0900 (Zeitgeber time 0; ZT 0). Immediately after the SD, the recovery period was recorded continuously for 18 hours. In order to estimate the phenotypic sleep pressure in mice, we measured the awaking threshold against external sensory stimuli in mice. Each mouse was set for sleep recording, and then a cage of mice was put on the shaker (rocking mixer; As One, Japan). Shaking was started 30 seconds after the appearance of EEG delta wave activity. The latency from the start of shaking to awaking determined by EEG and EMG was used as a parameter of sleepAugmented Sleep Pressure Model in MiceFigure 2. The influence of dietary restriction during gestation on sleep/wake architecture, body temperature, and spontaneous activity in adult offspring mice. Diurnal changes of wakefulness (A), NREM sleep (B), and REM sleep (C). Mean bin size (D), number of episodes (E), and mean interval (F) in each sleep/wake state for all 24-hour recordings. Hourly time course of body temperature (G) and spontaneous activity (H). Spontaneous activity was averaged for each 6-hour period (I) across ZT0-6 (L1), ZT6-12 (L2), Title Loaded From File ZT12-18 (D1), and ZT18-24 (D2). Open bars and circles indicate AD mice. Closed bars and circles indicate DR mice. Data represent means 6 SEM (A ; n = 6, G ; n = 5). *p,0.05 indicates a significant difference. doi:10.1371/journal.pone.0064263.gpressure. The procedure of shaking and latency measurement was performed 5 times and the latency was averaged for each mouse.bodies by enzymatic assay (Kainos Laboratories, Tokyo, Japan). Glucose was detected from tail blood by a glucose biosensor (LifeScan, Inc., Milpitas, CA, USA).Measurement of Plasma Levels of Triglycerides, Free Fatty Acids (FFAs), Ketone Bodies, and GlucoseAt the time of decapitation, trunk blood was coll.En to a computer-assisted data acquisition system CED 1401 data processor (CED, Cambridge, UK). After the habituation period, polygraphic recordings of body temperature, spontaneous activity, EEG, and EMG were collected continuously, throughout the L/D cycle. Telemetric signals of body temperature were sampled by Dataquest ART (Data Sciences Int., USA). Spontaneous activity was also recorded via signal strength of the telemetric device for body temperature. EEG and EMG signals were sampled by the Spike2 acquisition program (Cambridge Electronic Design, UK). Offline sleep scoring was first done via Spike2 software, and carefully verified by visual assessment of the EEG and EMG activities. Vigilance states were classified over 6-s epochs as wakefulness, NREM, or rapid eye movement (REM) sleep. NREM sleep was characterized by a continuous, slow, high-voltage EEG and low-voltage EMG activity. REM sleep was characterized by low-voltage EEG with continuous theta waves and total suppression of the EMG. Fast Fourier Transform using the Spike2 analysis program (Cambridge Electronic Design, UK) calculated the EEG power spectrum in the epoch determined to be NREM sleep. The sampling rate for EEG/EMG data collection was 100 Hz. The EEG delta and theta frequency band was set at 0.5?.0 Hz and 4.0?.0 Hz, respectively. Each value of power 16985061 was presented as a percentage of the total power (0.5?0 Hz). In this study, we used delta/theta value as a parameter of SWA [26]. Body temperature, spontaneous activity, sleep/wake durations, and EEG delta/theta ratio were averaged for hourly intervals. After the 24-hour baseline-recording period, 6-hour SD by gentle touching with a brush was carried out, starting at 0900 (Zeitgeber time 0; ZT 0). Immediately after the SD, the recovery period was recorded continuously for 18 hours. In order to estimate the phenotypic sleep pressure in mice, we measured the awaking threshold against external sensory stimuli in mice. Each mouse was set for sleep recording, and then a cage of mice was put on the shaker (rocking mixer; As One, Japan). Shaking was started 30 seconds after the appearance of EEG delta wave activity. The latency from the start of shaking to awaking determined by EEG and EMG was used as a parameter of sleepAugmented Sleep Pressure Model in MiceFigure 2. The influence of dietary restriction during gestation on sleep/wake architecture, body temperature, and spontaneous activity in adult offspring mice. Diurnal changes of wakefulness (A), NREM sleep (B), and REM sleep (C). Mean bin size (D), number of episodes (E), and mean interval (F) in each sleep/wake state for all 24-hour recordings. Hourly time course of body temperature (G) and spontaneous activity (H). Spontaneous activity was averaged for each 6-hour period (I) across ZT0-6 (L1), ZT6-12 (L2), ZT12-18 (D1), and ZT18-24 (D2). Open bars and circles indicate AD mice. Closed bars and circles indicate DR mice. Data represent means 6 SEM (A ; n = 6, G ; n = 5). *p,0.05 indicates a significant difference. doi:10.1371/journal.pone.0064263.gpressure. The procedure of shaking and latency measurement was performed 5 times and the latency was averaged for each mouse.bodies by enzymatic assay (Kainos Laboratories, Tokyo, Japan). Glucose was detected from tail blood by a glucose biosensor (LifeScan, Inc., Milpitas, CA, USA).Measurement of Plasma Levels of Triglycerides, Free Fatty Acids (FFAs), Ketone Bodies, and GlucoseAt the time of decapitation, trunk blood was coll.
