NEUROPHARMACOLOGY OF AROUSAL AND SLEEP DISORDERS

描述(申请人提供)：这项研究计划的主要目的是从神经药理学的角度研究唤醒和睡眠的细胞机制，特别强调发作性睡病的发病机制。将研究新发现的多肽神经递质--下克汀素/食欲素及其受体。下丘脑克汀素/食欲素及其受体在发作性睡病和睡眠障碍以及食物摄入量的调节中发挥作用。目前，人们对它们对脑神经元的生理影响知之甚少。以前的研究表明，脑核中的下丘脑肌红素/食欲素缺乏通过下丘脑肌红素/食欲素2受体产生的兴奋是发作性睡病的中心成分。拟议的项目侧重于通过分离的大鼠和小鼠大脑核的原代培养，进一步阐明它们在细胞和分子水平上的作用，这些核参与调节觉醒和睡眠。具体项目包括：(1)阐明下丘脑素/食欲素对结节乳头核内富含下丘脑素/食欲素2受体的组胺能神经元的影响；(2)研究下丘脑区去甲肾上腺素能神经元对富含下丘脑素/食欲素1受体的蓝斑核内去甲肾上腺素能神经元的影响；(3)研究下丘脑素/食欲素对Meynert基底核胆碱能神经元的影响。此外，一个异源系统(HEK293A) 将使用与每种类型的下丘脑素/增食欲素受体一起转染的受体。所有这些计划都侧重于阐明下丘脑肌红素/食欲素的信号转导机制，并确定G蛋白的身份和作用。将使用电生理技术(膜片钳)和药理技术与分子生物学方法相结合。这些项目对于加深对发作性睡病和其他睡眠障碍的理解非常重要。所获得的知识对于开发更有效的治疗发作性睡病和其他睡眠障碍的方法至关重要。这项关于大脑睡眠唤醒中心的研究将有助于帮助那些患有睡眠障碍的人。

DESCRIPTION (provided by applicant): The main objective of this research proposal is to investigate neuropharmacologically the cellular mechanisms of arousal and sleep with a special emphasis on the pathogenesis of narcolepsy. Hypocretins/orexins, newly discovered peptide neurotransmitters, and their receptors will be studied. Hypocretins/orexins and their receptors play a role in narcolepsy and sleep disorders and in the regulations of food intake. Currently little is known about their physiological effects on brain neurons. Previous studies have shown that lack of excitation produced by hypocretins/orexins through hypocretin2/orexin2 receptors in the brain nuclei is the central component of narcolepsy. Proposed projects focus on further elucidating their actions at the cellular and molecular level by using dissociated primary cultures of rat and mouse brain nuclei that are involved in regulating arousal and sleep. The specific projects are: (1) to elucidate hypocretin/orexin effects on histaminergic neurons in the tuberomammillary nucleus which are rich in hypocretin2/orexin2 receptors, (2) to determine the transmitter effects on noradrenergic neurons in the locus coeruleus which are rich in hypocretin1/orexin 1 receptors, and (3) to investigate hypocretin/orexin effects on cholinergic neurons in the nucleus basalis of Meynert. In addition, a heterologous system (HEK293A) which is transfected with each type of hypocretin/orexin receptor will be used. All of these projects emphasize elucidating signal transduction mechanisms of hypocretins/orexins, and determining the identity and roles of G proteins. Electrophysiological techniques (patchclamp) and pharmacological techniques combined with molecular biological methods will be used. These projects are important for deepening the understanding of narcolepsy and other sleep disorders. The knowledge obtained is essential in developing more effective treatments for narcolepsy and other sleep disorders. This research on the sleep-arousal centers of the brain will contribute in helping those who suffer sleep disorders.