フォワード・ジェネティクスで同定した新規睡眠制御遺伝子による分子機構の解明

利用正向遗传学方法，我们建立了几个突变小鼠谱系显示扰动的睡眠/觉醒行为。Sleepy 2突变增加了NREM睡眠时间，在黑暗阶段更为明显。值得注意的是，Sleepy 2基因家族的高度同源家族成员也表现出异常的睡眠/觉醒行为，包括睡眠需求的增加。为了了解睡眠稳态调节的分子和细胞机制，可以通过下一代测序来检查基因签名的变化。这项研究是在多重水平上设计的，以检查基因突变和睡眠剥夺的影响。睡眠需求是睡眠稳态维持的标志，因为它随着长时间的觉醒而增加。也可以检查睡眠剥夺后的反应，以评估睡眠/觉醒的正常维持。选择大脑的几个区域，并处理来自对称半球的样品用于RNA和ChIP测序。这种方法允许分析受研究设计应用的各种条件影响的转录组学和表观遗传变化。基于转录组学分析，大脑区域被缩小，以便对基因签名的变化进行更有力和仔细的研究。将进行广泛的数据整合，以绘制相互作用的分子和信号转导途径。这将补充目前可用的具有睡眠/觉醒调节特定机制的通路数据库。

Using the forward genetics approach, we established several mutant mice pedigree showing perturbed sleep/wake behavior. Sleepy2 mutant increase in the NREM sleep time, more pronounced during the dark phase. Notably, highly homologous family members of Sleepy2 gene family also showed aberrant sleep/wake behavior including the increase in sleep need.In order to understand the molecular and cellular mechanism of sleep homeostasis regulation, changes in gene signature can be examined by the next-generation sequencing. The study was designed at multiplexed level to examine both the effect of the gene mutation and sleep deprivation. Sleep need is a marker for the sleep homeostasis maintenance as it increases with prolonged wakefulness. The response following sleep deprivation can also be examined to assess normal maintenance of sleep/wakefulness. Several regions of the brain were selected and the samples from symmetrical hemisphere were processed for RNA- and ChIP-sequencing. This approach allows analysis of both transcriptomic and epigenetic changes affected by the various conditions applied by the study design.Based on the transcriptomics analysis, a brain region is narrowed down for more vigorous and scrutinized study for the changes in gene signature. An extensive data integration will be performed to map the interacting molecules and signal transduction pathway. This will supplement the currently available pathway database with sleep/wake regulation specific mechanism.