Regulation of Sleep and Circadian Behaviors by Prokineticin 2

类睡眠状态在整个进化过程中都是保守的，慢性睡眠障碍影响着超过10%的人类。然而，调节睡眠的遗传和神经机制仍然知之甚少。大多数睡眠研究都使用了夜间活动的哺乳动物，如老鼠和大鼠，但尚不清楚昼夜节律方面的睡眠调节是否适用于人类等白天活动的哺乳动物。我将利用斑马鱼相对简单但保守的神经回路和光学透明度来测试几个关于原激动素2在调节睡眠和昼夜行为中的作用的假说。斑马鱼是一种昼夜活动的脊椎动物。首先，我将通过使用高通量运动活动和觉醒阈值分析进行功能获得和功能丧失的遗传学实验，来验证Prokineticin 2作为昼夜节律信号抑制运动活动的假设。其次，我将使用一种生物发光报告来检验斑马鱼幼体中原激动素2神经元在夜间活动的假设，该生物发光报告监测行为自由的斑马鱼幼体中特定于基因的神经元的活动。第三，我将通过监测这些神经元群体对原激动素2过度表达的反应来检验这样的假设，即原激动素2抑制促进觉醒的神经元，激活促进睡眠的神经元。这些实验有可能阐明调节昼夜节律输出和调节睡眠的神经机制，这可能会导致睡眠和昼夜节律相关疾病的新疗法。

Sleep-like states are conserved throughout evolution, and chronic sleep disorders affect over 10% of humans. However, the genetic and neural mechanisms that regulate sleep remain poorly understood. Most sleep studies have used nocturnal mammals such as mice and rats, but it is unclear whether circadian aspects of sleep regulation are conserved with diurnal mammals such as humans. I will exploit the relatively simple yet conserved neural circuits and optical transparency of the zebrafish, a diurnal vertebrate, to test several hypotheses regarding the role of Prokineticin 2 in regulating sleep and circadian behaviors. First, I will test the hypothesis that Prokineticin 2 acts as a circadian signal to inhibit locomotor activity by performing gain and loss of function genetic experiments using high throughput locomotor activity and arousal threshold assays. Second, I will test the hypothesis that Prokineticin 2 neurons are active at night in zebrafish larvae using a bioluminescence reporter that monitors the activities of genetically specified neurons in freely behaving zebrafish larvae. Third, I will test the hypothesis that Prokinectin 2 inhibits wake-promoting neurons and activates sleep-promoting neurons by monitoring the activities of these neural populations in response to Prokineticin 2 overexpression. These experiments have the potential to clarify the neural mechanisms that mediate circadian output and regulate sleep, which may lead to new therapies for sleep and circadian related disorders.