This is challenging because REM sleep is a relatively rare state compared with NREM sleep and wake and because episodes of REM sleep are short. The goal of this study was, therefore, to determine whether we could develop an algorithm to identify REM vs NREM sleep in mice based on digital video recordings. Subtle differences among the 3 states can be detected visually. A time-series plot of mean aspect ratio for 1 mouse with colors corresponding to the manually scored state (WAKE, non-rapid eye movement (NREM), and rapid eye movement (REM) sleep). This, we believe, is related to the mouse becoming more atonic in REM sleep. In particular, the area and aspect ratio of the mouse, respectively, increased and decreased when the mouse went from NREM to REM sleep (see Figure 1). Nonetheless, our video recordings show a subtle signal for REM sleep. 5Īlthough these methods are quite accurate to determine wake and sleep, they cannot distinguish non-rapid eye movement sleep (NREM) from rapid eye movement sleep (REM). The other strategy is based on piezoelectric detection of mouse movements by pressure sensors in the floor of the mouse cage analysis of the data recorded by such sensors reveals patterns that are characteristic of sleep and wakefulness.
ELUCIDATE 5AM TRACK LIST MANUAL
This approach, termed the 40-second Rule, has been validated by comparison with manual scores based on EEG/EMG recordings in both young 4 and old 6 C57BL/6J mice. 4, 5 One approach is based on determining inactivity either by electronic beam splits or by video analysis any duration of inactivity that lasts 40 seconds or more is considered sleep. Two approaches to high-throughput phenotyping have already been proposed. in response to sleep, wake, and sleep deprivation and (2) to screen the large panel of knockout mice that have already been created. This strategy can be used (1) in studies that evaluate changes in mRNA, protein, etc. We seek an alternative high-throughput strategy that will obviate the need for EEG/EMG recording. Thus, the necessity for surgery, time to recover from surgery, and scoring of large numbers of epochs adds expense and makes studies of sleep in mice very labor intensive. Moreover, scoring of EEG/EMG records is labor intensive: if states are assessed in 10-second epochs across a 24-hour period, there are 8,640 epochs to be scored, whereas with 4-second epochs, there are 21,600 epochs. This technique requires surgery on mice with the implantation of electrodes mice cannot be studied until they recover from the surgery. 3 Use of these new mouse resources requires evaluation of the effect of knockout of a specific gene on sleep and its substages as well as wakefulness.Ĭurrently, this evaluation is performed by assessing changes in the electroencephalogram (EEG) and electromyogram (EMG). There are currently large numbers of mice available with knockout of specific genes. Transgenic strategies typically create altered gene function on a C57BL/6J background by using selective breeding strategies to minimize the effects of genetic background. These include the analysis of changes in the transcriptome with sleep, wake, and sleep deprivation (for review, see 2) as well as the use of specific transgenic mice. Multiple strategies are used to identify relevant genes (for review, see 1). A major focus of current research in mice is to elucidate gene products that (1) regulate sleep and wake, (2) are regulated by sleep and wake, or (3) are affected by sleep deprivation.
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