Cellular and Molecular Mechanisms of REM Sleep Control

说明(申请人提供)：胆碱能系统，特别是 背外侧核和桥脑脚外侧核(LDT/PPT)细胞群 在桥脑-中脑被盖(PMT)内，积极参与 快速眼动(REM)睡眠的时间和数量。我们的初步研究 导致了两个重要的发现：1)胆碱诱导的长期 增强快速眼动睡眠(LTRE)：和2)长期增强 脑桥-膝枕(PGO)波(LTPE)是一个明显的阶段 REM的组件。本提案所述研究的重点是 确定PMT中的哪些神经元表现出独特的神经元模式 与单点源注射胆碱能有关的活性 激动剂卡巴胆碱进入尾侧PMT，产生这些长期的 效果。我们的假设是，脑内胆碱能部位的激活 尾侧PMT改变REM睡眠和PGO的突触后兴奋性 通过刺激即刻早期基因的延长表达来建立网络 强化了这些长期影响。这些实验将测试我们的模型 尾侧PMT作为REM调节区的研究 我开发的用于标记传入的荧光纳米球探针 药物刺激大脑后的神经元。第二，模式 在这些过程中，PMT中Fos的表达证明了神经元的激活 将对长期影响进行考察。第三，对LTRE/LTPE的影响如下 抑制PMT中Fos蛋白的研究将通过以下方式进行 微量注射c-fos反义寡核苷酸。最后，这些联系 在LDT/PPT神经元被激活的情况下，它们包含的神经递质， C-fos、fos B、jun-B和EGR-1基因的时空分布 它们表达的蛋白质和CREB介导的基因的信号诱导 转录将使用顺行免疫组织化学(FITC)进行研究 (葡聚糖)和逆行(罗丹明纳米球)标记。这些措施的结果 实验应该为理解细胞和分子提供基础。 PMT尾侧区神经元激活神经元网络的机制 参与REM睡眠和PGO活动的长期维持。这个 暗示从睡眠扩展到所有适应行为状态，在这些状态下 神经系统必须以统一的方式作出反应，以重置其激活水平 以及大脑某一部分的急性突触事件如何引发变化 在偏远地点的基因产物中。

DESCRIPTION (provided by applicant): The cholinergic system, particularly the cell groups of the laterodorsal and pedunculopontine tegmental (LDT/PPT) nuclei within the ponto-mesencephalic tegmentum (PMT), is actively involved in the timing and quantity of rapid eye movement (REM) sleep. Our preliminary studies have led to two important discoveries: 1) cholinergically induced long-term enhancement of REM sleep (LTRE): and 2) long-term enhancement of ponto-geniculo-occipital (PGO) waves (LTPE) that are a distinguishing phasic component of REM. The focus of the studies described in this proposal is to determine which neurons in the PMT demonstrate unique patterns of neuronal activity in association with a single point source injection of the cholinergic agonist carbachol into the caudolateral PMT that produces these long-term effects. Our hypothesis is that cholinergic activation of sites in the caudolateral PMT alters the postsynaptic excitability of the REM sleep and PGO networks by stimulating the prolonged expression of immediate-early genes that potentiates these long-term effects. The experiments will test our model of the caudolateral PMT as a REM regulatory region by using the carbachol conjugated fluorescent nanosphere probe that I have developed for labeling afferent neurons following pharmacological stimulation of the brain. Second, the pattern of activation of neurons as evidenced by Fos expression in the PMT during these long-term effects will be examined. Third, the effects on LTRE/LTPE following suppression of Fos protein in the PMT will be studied by performing microinjections of c-fos antisense oligonucleotides. Finally, the connections with LDT/PPT neurons that are activated, the neurotransmitters they contain, the temporal and spatial distribution of c-fos, fos B, jun-B, and egr-1 gene proteins they express, and the signaling induction of CREB-mediated gene transcription will be studied using immunohistochemistry with anterograde (FITC dextran) and retrograde (rhodamine nanospheres) labeling. The outcome of these experiments should provide a basis for understanding the cellular and molecular mechanisms by which neurons in the caudolateral PMT activate neuronal networks involved in the long-lasting maintenance of REM sleep and PGO activity. The implications extend beyond sleep to all adaptive behavioral states in which neuronal systems must react in a unified way to reset their activation levels and to how acute synaptic events in one part of the brain trigger alterations in gene product at remote sites.