Circadian clock protein Period3 contributes to sleep Homeostasis through GABA signaling in zebrafish
作者单位：Soochow University
会议名称：《中国睡眠研究会第十一届全国学术年会》
会议日期：2019年
摘 要：The zebrafish(Danio rerio) has figured prominently as a vertebrate model for studying circadian clocks and sleep. Zebrafish are a strong model for developmental biology research because of their fecundity, larval-stage transparency, short time to hatching and ease in handling;furthermore, they are well characterized in terms of development, neurobiology, and genetics. Moreover, there are many experimental tools not easily applied in mammalian models that can be readily applied to zebrafish. It is known that both the homeostatic process(S) and the circadian process(C) contribute to regulation of sleep homeostasis. While we have a good understanding of circadian regulation, relatively little is known about molecular mechanisms underlying circadian regulation of sleep homeostasis and/or interaction between the circadian clock system and sleep. A variable-number(4 or 5) tandem-repeat polymorphism in the coding region of the clock gene PERIOD3 was shown to confer vulnerability to sleep loss and circadian misalignment through its effects on sleep homeostasis in human beings. While Per3 knockout mice shows little impacts in circadian functions and sleeping behaviors. Here we reported characterization of zebrafish period3(per3) null mutants generated by TALEN method. Locomotor assays showed that per3 mutant fish display 1.1-hour shortened period and approximately 3-hour phase advance compared with wild types fish under constant dark condition, and are nearly 3 hours shortened period under constant light condition;and also bmal1b, per1a, per1b and per2 are down-regulated in per3 mutant fish;Deep sequencing-based transcriptome analysis also showed a lot of circadian genes were altered in per3 mutant fish, indicating that per3 is essential for zebrafish circadian regulation. Cell transfection experiments showed that zebrafish Per3 can regulate the expression of bmal1 b through RRE box. Intriguingly, per3 mutant fish display less sleep time in light dark condition, decreased ar