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

阐明正向遗传学鉴定的新型睡眠调节基因的分子机制

• 批准号: 17J07957
• 负责人: KIM StaciJakyong
• 金额: $ 1.09万
• 依托单位: University of Tsukuba
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for JSPS Fellows
• 财政年份: 2017
• 资助国家: 日本
• 起止时间: 2017-04-26 至 2019-03-31
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-17J07957/
• 关键词: transcriptomics; forward genetics; mouse genetics; sleep; Forward genetics; Sleep

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.

利用正向遗传学方法，我们建立了几个表现出扰乱睡眠/觉醒行为的突变小鼠家系。Sleepy2突变体增加了NREM睡眠时间，在黑暗阶段表现得更为明显。值得注意的是，Sleepy2基因家族高度同源的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。睡眠需求是维持睡眠动态平衡的标志，因为睡眠需求随着清醒时间的延长而增加。还可以检查睡眠剥夺后的反应，以评估睡眠/清醒的正常维持。选择了大脑的几个区域，并对来自对称半球的样本进行了RNA和芯片测序。这种方法可以分析受研究设计应用的各种条件影响的转录和表观遗传变化。基于转录切分分析，缩小大脑区域，以便更有力和仔细地研究基因签名的变化。将进行广泛的数据集成，以绘制相互作用的分子和信号转导路径。这将补充目前可用的通路数据库，提供睡眠/觉醒调节的特定机制。

项目成果

Forward genetics study identifies novel gene in sleep/wake regulation

正向遗传学研究发现了睡眠/觉醒调节中的新基因

• 发表时间: 2018
• 作者: Staci Jakyong Kim;Tomoyuki Fujiyama;Chika Miyoshi;Mizuno Seiya;Noriko Hotta-Hirashima;Satomi Kanno;Aya Ikkyu;Satoru Takahashi;Hiromasa Funato;Masashi Yanagisawa
• 通讯作者: Masashi Yanagisawa

Identification of a novel gene family in sleep and wake regulation

睡眠和觉醒调节中新基因家族的鉴定

• 发表时间: 2018
• 作者: Staci Jakyong Kim;Tomoyuki Fujiyama;Chika Miyoshi;Mizuno Seiya;Noriko Hotta-Hirashima;Satomi Kanno;Aya Ikkyu;Satoru Takahashi;Hiromasa Funato;Masashi Yanagisawa
• 通讯作者: Masashi Yanagisawa

Brain-specific conditional knockout of Sleepy2 alters sleep and wake behavior

Sleepy2 的大脑特异性条件性敲除会改变睡眠和醒来行为

• 发表时间: 2018
• 作者: Staci Jakyong Kim;Tomoyuki Fujiyama;Chika Miyoshi;Noriko Hotta-Hirashima;Satomi Kanno;Aya Ikkyu;Hiromasa Funato;Masashi Yanagisawa
• 通讯作者: Masashi Yanagisawa