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，我们建立了几种表现出睡眠/觉醒行为紊乱的突变小鼠系谱。Sleepy2突变体在非快速眼动睡眠时间增加，在黑暗期更为明显。值得注意的是，高度同源的Sleepy2基因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测基因标记的变化。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。睡眠需求随着清醒时间的延长而增加，是维持睡眠稳态的标志。睡眠剥夺后的反应也可以通过检查来评估睡眠/清醒的正常维持。选择大脑的几个区域，并从对称半球的样本进行RNA和芯片测序处理。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，以便对基因特征的变化进行更有力、更仔细的研究。广泛的数据整合将执行映射相互作用的分子和信号转导途径。这将补充目前可用的睡眠/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.