Genetic Approaches to Sleep/Wake and Response to Sleep Loss in Mice

描述(由申请者提供)：睡眠不足在美国人中很常见，而且越来越普遍。失眠会带来一系列的不良后果。这些包括行为变化、认知障碍和机动车撞车风险增加，以及工作场所的错误。此外，睡眠不足会导致代谢变化，导致胰岛素抵抗，食欲增加，是肥胖发展的风险因素。然而，睡眠不足造成的损害程度有很大的个体差异。这是可遗传的，也就是说，个体之间的差异很大程度上是由卵黄蛋白基因决定的。阐明睡眠缺失反应的遗传基础将使需要全天候活动的手术安排更加合理，并可能识别新的途径，可能成为未来改变睡眠缺失后果的药物方法的基础。然而，进一步研究人类涉及的基因是具有挑战性的，因为检查对睡眠剥夺的反应需要昂贵的研究和高协议负担(实验室中的四天)。在这项应用中，基于对小鼠的研究，提出了一种识别负责基因的替代方法。不同近交系小鼠对睡眠剥夺的反应各不相同，但造成这种差异的基因变异尚不清楚。这项提案计划利用最近培育出的基因完全不同的多样化的近交系小鼠。这一策略需要对大量小鼠进行研究。 (在本应用中为800)以高吞吐量方式。因此，对睡眠剥夺的反应将通过一种基于数字视频分析的新的高通量策略来评估。视频分析不仅提供对睡眠和醒来的准确估计，而且还提供对睡眠阶段的准确估计--快速眼动(REM)睡眠和非快速眼动(NREM)睡眠。所有小鼠不仅将进行表型分析，还将基于一种新的具有7000个单核苷酸多态的基因分型芯片进行基因分型。这种方法允许识别与这种数量性状相关的小鼠基因组的一小部分。此外，还将同时评估一只小鼠可以维持的正常清醒时间。这也是一种可遗传的特征， 与常见的睡眠困难问题高度相关。对于这两个数量性状，将在来自协作杂交系的相关小鼠中进行验证研究。未来的研究还将使用具有良好特征的个体样本将这项调查扩展到人类群体，这些个体已经被研究过睡眠缺失，并且他们的DNA已经可以获得。

DESCRIPTION (provided by applicant): Sleep deprivation is common and increasing in prevalence in the American population. Loss of sleep has a number of adverse consequences. These include behavioral changes, impairment of cognition with an increased risk of motor vehicle crashes, and errors in the workplace. Moreover, sleep loss leads to metabolic changes with insulin resistance, increased appetite and is a risk factor for development of obesity. There are, however, substantial individual differences in the degree of impairment produced by sleep loss. This is heritable, i.e., a large part of the difference between individuals is genetic in oriin. Elucidating the genetic basis of the response to sleep loss will enable a more rationale scheduling in operations that require 24/7 activity, and will likely identify novel pathways that could be the basis of future pharmacological approaches to alter the consequences of sleep loss. Further study to elucidate genes involved in humans is, however, challenging since examination of the response to sleep deprivation requires expensive studies with a high protocol burden (four days in a laboratory). In this application an alternative approach to identify responsible genes is proposed, based on studies in mice. Response to sleep deprivation in mice varies between inbred strains but the gene variants responsible for this difference are unknown. This proposal plans to take advantage of the recently created diverse outbred strain of mice which are all genetically different. This strategy requires study of a large number of mice (in this application, 800) in a high throughput fashion. Hence, response to sleep deprivation will be assessed by a novel high throughput strategy based on digital video analysis. Video analysis not only provides accurate estimates of sleep and wake, but also of the stages of sleep-rapid-eye movement (REM) sleep and non-rapid-eye movement (NREM) sleep. All mice will not only be phenotyped but will be genotyped based on a new genotyping chip with 7,000 single nucleotide polymorphisms. This approach allows identification of a small region of the mouse genome associated with this quantitative trait. In addition, the normal duration of wakefulness that a mouse can sustain will also be simultaneously assessed. This, too, is a heritable trait that is highly relevant to the common problem of sleeping difficulty. Validation studies in relevant mice from the collaborative cross lines will be employed for both quantitative traits. Future studies will also extend this investigation into human populations using samples of well characterized individuals who have been studied with sleep loss and whose DNA is already available.