新規コンディショナル過眠モデルマウスを用いた睡眠制御分子メカニズムの解明

使用新的条件性嗜睡模型小鼠阐明睡眠控制分子机制

• 批准号: 21K15004
• 负责人: KIM Staci・Jakyong
• 金额: $ 2.91万
• 依托单位: University of Tsukuba
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Early-Career Scientists
• 财政年份: 2021
• 资助国家: 日本
• 起止时间: 2021-04-01 至 2024-03-31
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-21K15004/
• 关键词: Sleep; Intracellular signaling; Sleep depth; Sik3; Gene-modified mice

There has been increased interest in understanding how sleep/wake behavior is controlled as the emphasis on the well-maintained sleep has grown. However, the regulatory mechanism of sleep and wake at the molecular level remains unclear to date. This study is based on our previous finding of SIK3 kinase as a key regulator of sleep homeostasis. We identified several SIK3 substrate molecules where SIK3-induced phosphorylation of these proteins localizes them to the cytoplasm and result in desuppression of target gene expression in the nucleus. We observed abnormal sleep and wake behavior in the loss-of-function mutants and phosphodeficient SIK3-substrate mice. The loss-of-function mutants showed increased sleep time and sleep need, consistent with the Sik3-Sleepy mutant phenotype. On the other hand, the phosphodeficient mutants showed increased wakefulness with decrease in sleep need marker. These results suggest that the SIK3 cascade may constitute an important molecular pathway in sleep need regulation. To investigate responsible neural population in sleep regulation, we established floxed mice to induce conditional knockout of the target gene when mated with Cre-driver mice. It is plausible that sleep time and sleep need regulation may be controlled by distinct population of neurons and cell types. Several Cre-driver strains were selected to mate with the floxed mice and examined for changes in any sleep parameters. We are proceeding with investigation of the role of SIK3 substrates in brain region- and cell type-specific manner using these newly established mouse line.

随着人们对良好睡眠的重视，人们对了解如何控制睡眠/觉醒行为的兴趣越来越大。然而，迄今为止，在分子水平上对睡眠和觉醒的调节机制仍不清楚。这项研究是基于我们以前发现的SIK3激酶作为睡眠稳态的关键调节因子。我们鉴定了几种SIK3底物分子，其中SIK3诱导的这些蛋白质的磷酸化将它们定位于细胞质并导致细胞核中靶基因表达的去抑制。我们观察到异常的睡眠和觉醒行为的功能丧失突变体和磷酸缺陷的SIK3底物小鼠。功能丧失突变体显示睡眠时间和睡眠需求增加，与Sik3-Sleepy突变体表型一致。另一方面，磷酸缺陷突变体显示觉醒增加，睡眠需要标记物减少。这些结果表明，SIK3级联可能构成了睡眠需要调节的重要分子途径。为了研究睡眠调节中的神经元群体，我们建立了floxed小鼠，当与Cre-driver小鼠交配时诱导靶基因的条件性敲除。睡眠时间和睡眠需求调节可能由不同的神经元和细胞类型控制，这是合理的。选择几种Cre-驱动品系与floxed小鼠交配，并检查任何睡眠参数的变化。我们正在使用这些新建立的小鼠系研究SIK3底物在脑区域和细胞类型特异性方式中的作用。

项目成果

Changes in sleep/wake by acute disruption in Hdac4 signaling

Hdac4 信号传导的急性破坏导致睡眠/觉醒的变化

• 发表时间: 2022
• 作者: Staci Kim;Chika Miyoshi;Hiromasa Funato;et al.
• 通讯作者: et al.

Intracellular signaling components in homeostatic regulation of sleep

睡眠稳态调节中的细胞内信号成分

• 发表时间: 2022
• 作者: Staci J. Kim;Shinya Nakata;Noriko Hotta-Hirashima;Nodoka Aasama;Aya Ikkyu;Miyo Kakizaki;Satomi;Seiya Mizuno;Satoru Takahashi;Chika Miyoshi;Hiromasa Funato;Masasyi Yanagisawa
• 通讯作者: Masasyi Yanagisawa

Kinase signalling in excitatory neurons regulates sleep quantity and depth

• DOI: 10.1038/s41586-022-05450-1
• 发表时间: 2022-12-07
• 期刊: NATURE
• 影响因子: 64.8
• 作者: Kim，Staci J.;Hotta-Hirashima，Noriko;Funato，Hiromasa
• 通讯作者: Funato，Hiromasa