Identification of preoptic sleep neurons using retrograde labelling and gene profiling.
Identification of preoptic sleep neurons using retrograde labelling and gene profiling.
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DOI：
10.1038/nature22350
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发表时间：
2017-05-25
期刊：
影响因子：
64.8
通讯作者：
Dan Y
中科院分区：
文献类型：
--
作者：
Chung S;Weber F;Zhong P;Tan CL;Nguyen TN;Beier KT;Hörmann N;Chang WC;Zhang Z;Do JP;Yao S;Krashes MJ;Tasic B;Cetin A;Zeng H;Knight ZA;Luo L;Dan Y
在人类和其他哺乳动物物种中，下丘脑视前区（POA）的病变会导致严重的睡眠障碍，表明POA在睡眠产生中起着至关重要的作用。然而，潜在的电路机制仍然知之甚少。电生理记录和c-Fos免疫组化结果显示，POA内存在睡眠神经元，尤其是腹外侧视前区（VLPO）和正中视前核（MnPO）。已显示c-Fos标记的睡眠活性神经元的药物遗传学激活可诱导睡眠。然而，睡眠活跃神经元在空间上与唤醒活跃神经元混合，使得难以专门针对睡眠神经元进行电路分析。在这里，我们已经确定了一个群体的POA睡眠神经元的基础上，他们的投射目标，并发现了他们的分子标记。利用表达通道视紫红质2（ChR 2）或光激活氯离子通道（iC++）的慢病毒进行逆行标记、双向光遗传学操作和光电极记录，我们发现投射到结节乳头核（TMN）的POA GABA能神经元既具有睡眠活性又具有睡眠促进作用。此外，翻译核糖体亲和纯化（TRAP）和单细胞RNA-seq鉴定了这些神经元的候选标记物，并且光遗传学和药物遗传学操作证明了几种肽标记物（胆囊收缩素、促肾上腺皮质激素释放激素和速激肽1）标记促进睡眠的神经元。总之，这些发现为促进睡眠的POA神经元提供了简单的遗传途径，并为解剖睡眠控制回路提供了有价值的切入点。
In humans and other mammalian species, lesions in the preoptic area (POA) of the hypothalamus cause profound sleep impairment, indicating a crucial role of the POA in sleep generation. However, the underlying circuit mechanism remains poorly understood. Electrophysiological recordings and c-Fos immunohistochemistry showed the existence of sleep-active neurons in the POA, especially in the ventrolateral preoptic area (VLPO) and median preoptic nucleus (MnPO). Pharmacogenetic activation of c-Fos-labeled sleep-active neurons has been shown to induce sleep. However, the sleep-active neurons are spatially intermingled with wake-active neurons, making it difficult to target the sleep neurons specifically for circuit analysis. Here, we have identified a population of POA sleep neurons based on their projection target and discovered their molecular markers. Using a lentivirus expressing channelrhodopsin-2 (ChR2) or a light-activated chloride channel (iC++) for retrograde labeling, bidirectional optogenetic manipulation, and optrode recording, we showed that the POA GABAergic neurons projecting to the tuberomammillary nucleus (TMN) are both sleep active and sleep promoting. Furthermore, translating ribosome affinity purification (TRAP) and single-cell RNA-seq identified candidate markers for these neurons, and optogenetic and pharmacogenetic manipulations demonstrated that several peptide markers (cholecystokinin, corticotropin releasing hormone, and tachykinin 1) label sleep-promoting neurons. Together, these findings provide easy genetic access to sleep-promoting POA neurons and a valuable entry point for dissecting the sleep control circuit.
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