A HIGH-THROUGHPUT ASSAY FOR GENETIC STUDIES OF SLEEP

觉醒状态是整体神经系统功能的一个基本方面;然而，由于EEG分析的困难，很少在基因改变的小鼠中进行睡眠和觉醒的测量。我们建议开发一种高通量的替代脑电图记录在小鼠中，这将大大促进睡眠和觉醒的测量。 我们的系统利用一个简单的压电换能器，包括动物休息的柔性地板。 呼吸或其他运动引起的地板扩张产生电信号。 在慢波睡眠期间，有节奏的呼吸会产生一致的、有规律的电信号模式。 快速眼动睡眠由于更不规则的呼吸而产生不太一致的信号，而唤醒产生由各种运动引起的显着不规则的模式。 因此，可以准确地确定所有三种警戒状态。 我们已经在年轻的老鼠身上验证了这个系统。 由于不需要手术，并且信号模式比EEG产生的信号模式更简单，因此对唤醒状态进行高通量表型分析的技术将相对容易。 对我们系统的直接测试和使用将是调查睡眠参数的基础基因。 双胞胎研究、收养研究和对近交系小鼠的分析表明，睡眠相关性状的变化在很大程度上取决于基因型。 具体来说，我们建议通过比较我们的压电记录与传统的EEG记录，以验证我们的系统在常见的近交系小鼠。 基于我们先前的EEG数据，我们期望证实AKR/J和DBA/2 J近交系之间存在较大的睡眠差异。 随后将检查AKD 2F 1杂种和AKXD重组近交系。 这些数据的QTL分析应确定候选基因座，这两个菌株的睡眠性状的变异性的基础。 利用这种高通量系统，我们还将能够检查来自这些或其他菌株之间的适当遗传杂交的大量后代，以确认推定的QTL。 该系统还使大规模筛选诱变小鼠变得更加可行，我们将开始在拟定研究中检查此类小鼠。 总的来说，我们提出的项目将提供神经系统功能的重要指标，特别是将允许遗传方法来理解睡眠和睡眠障碍，这在现有技术下是不可能的。

Arousal states are a fundamental aspect of global nervous system function; yet, measures of sleep and wake are rarely performed in genetically altered mice due to the difficulty of EEG analysis. We propose to develop a high-throughput alternative to EEG recordings in mice that will greatly facilitate the measurement of sleep and wake. Our system utilizes a simple piezoelectric transducer comprising the flexible floor upon which the animal rests. Distension of the floor by respiratory or other movements produces electrical signals. During slow wave sleep, rhythmic breathing results in a consistent, regular pattern of electical signals. REM sleep produces a less consistent signal due to more irregular breathing, while wake produces a dramatically irregular pattern caused by a variety of movements. Therefore, all three vigilance states can be accurately determined. We have already validated this system in young rats. Since no surgery is required and the signal patterns are simpler than those produced by EEG, the technology to perform high-throughput phenotyping of arousal states will be relatively easy. An immediate test and use of our system will be to investigate genes that underlie sleep parameters. Twin studies, adoption studies, and the analysis of inbred strains of mice have demonstrated that variation in sleep related traits are largely determined by genotype. Specifically, we propose to validate our system in common inbred strains of mice by comparing our piezo recordings with traditional EEG recordings. Based on our prior EEG data, we expect to confirm large sleep differences between AKR/J and DBA/2J inbred strains. This will be followed by examination of AKD2F1 hybrids and AKXD recominant inbred lines. QTL analysis of these data should identify candidate loci that underlie the variability of sleep traits in these two strains. With this high throughput system we will also be able to examine large numbers of progeny from appropriate genetic crosses between these or other strains to confirm putative QTLs. This system also makes large scale screening of mutagenized mice far more feasible, and we will begin to examine such mice in the proposed study. In general, the project we propose will provide an important index of nervous system function, and specifically will allow genetic approaches towards understanding sleep and sleep disorders that would not be possible with existing technology.