Integrated research for the mechanism of sleep/wakefulness regulation

睡眠/觉醒调节机制的综合研究

基本信息

批准号：
20680021
负责人：
YAMANAKA Akihiro
金额：
$ 13.23万
依托单位：
National Institute for Physiological Sciences
依托单位国家：
日本
项目类别：
Grant-in-Aid for Young Scientists (A)
财政年份：
2008
资助国家：
日本
起止时间：
2008 至 2010
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-20680021/
关键词：
睡眠覚醒インビボトランスジェニックマウスチャネルロドプシン2 細胞外記録オレキシン視床下部スライスパッチクランプ脳波筋電図パッチクランプ遺伝子改変マウス緑色蛍光タンパク質光感受性タンパク質ナルコレプシー

项目摘要

Sleep/wakefulness is a physiological phenomenon which observed only in the whole body. Thus, it is essential to study using whole animal. Optogenetics is a recently developed technique to control the activity of neurons by expressing light activated proteins, such as channelrhodopsin2(ChR2) or halorhodopsin(Halo). This technique allows us to control the activity of orexin-producing neurons(orexin neurons) using light in vivo and in vitro. First, we confirmed the function of CHR2 and Halo in the orexin neurons using slice patch clamp and decided a condition of light illumination. Next, we proceeded in vivo experiment using conscious mouse. Optical fibers were bilaterally implanted into the hypothalamus. The activity of orexin neurons was inhibited by illuminating orange light into the hypothalamus through the optical fibers. At the same time, the activity of serotonergic neurons in the raphe nucleus is recorded by the extracellular recordings. Acute inhibition of orexin neurons induced slow wave sleep and the activity of serotonergic neurons was decreased in accordance with initiation of slow wave sleep. This result showed that inhibition of orexin neurons induced slow wave sleep in mice.

睡眠/清醒是一种生理现象，仅在整个身体中观察到。因此，使用全动物研究至关重要。光遗传学是一种最近开发的技术，可通过表达光活化的蛋白质（例如ChannelRhopopsin2（Chr2）或卤代紫红蛋白（Halo）（Halo）来控制神经元的活性。该技术使我们能够使用体内和体外光控制产生Orexin的神经元（Orexin神经元）的活性。首先，我们使用切片贴片夹确认了Orexin神经元中Chr2和Halo的功能，并确定了光照射的条件。接下来，我们使用有意识的小鼠进行了体内实验。将光纤植入下丘脑。通过光纤将橙色的光照射到下丘脑中，抑制了Orexin神经元的活性。同时，raphe核中血清素能神经元的活性由细胞外记录记录。根据慢波睡眠的开始，急性抑制Orexin神经元诱导的慢波睡眠，血清素能神经元的活性降低。该结果表明，抑制Orexin神经元诱导小鼠慢波睡眠。