睡眠-觉醒周期是最为显著的生物节律，然而生物钟在睡眠中调控的遗传机制尚不清楚。申请人前期通过对斑马鱼period3突变体的分析发现缺失Per3导致显著的睡眠障碍。本项目旨在阐明Per3对睡眠调控的遗传机制及其睡眠功能的演化规律：1）通过建立和检验per3驱动的GFP、Luciferase和GCamp6的转基因品系确定其在大脑中不同区域表达、振荡和细胞活动的特征；2）通过建立全身和组织特异基因敲除和过表达、光遗传学介导的神经元激活、化学遗传介导神经元杀死和高通量行为追踪分析等技术确定Per3是否通过γ-胺基丁酸（GABA）和去甲肾上腺素（Norepinephrine）信号在睡眠中起作用；3）通过建立和检验在正常和per3纯合突变体斑马鱼中表达人和小鼠正常PER3和携带突变的PER3的品系，确定生物钟蛋白Per3睡眠功能的演化规律。本研究将系统地阐明Per3对睡眠调节的遗传机制。

While the sleep-wake cycle is the most significant circadian rhythm, the molecular genetic mechanisms underlying how the circadian clock regulates sleep are still unclear. By the analysis and examination of the TALEN-generated period3 mutant zebrafish, the Han Wang laboratory recently found that loss of Per3 results in significant sleep disorders. The objective of this project is to elucidate the genetic mechanisms underlying Per3 regulation of sleep and the evolution of Per3 sleep function in three specific aims: 1) to establish and examine per3-driven GFP, Luciferase, and GCamp6 transgenic lines to determine its expression, rhythmicity and cellular activities in different regions of the brain; 2) to determine whether Per3 contributes to sleep via γ-aminobutyric acid (GABA) and norepinephrine signaling through generating whole-body and tissue-specific gene knockout and overexpression zebrafish lines, optogenetics-mediated neuronal activation, chemogenetics-mediated neuronal ablation as well as high-throughput behavioral recording and analytic techniques; and 3) to determine the evolution of Per3 sleep function by establishing and examining overexpression of wild-type and mutated PER3 of human and mouse in wild-type and per3 homozygous zebrafish, respectively. This study should systematically elucidate the genetic mechanisms of Per3 regulation of sleep.