Cellular and Molecular Mechanisms of REM Sleep Control

描述（由申请人提供）：胆碱能系统，特别是 背外侧和脚桥被盖（LDT/PPT）核的细胞群 在脑桥中脑被盖（PMT），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）胆碱能诱导的长期 增强REM睡眠（LTRE）：和2）长期增强 桥膝枕（PGO）波（LTPE）是一种区分相位 REM的组成部分。本提案所述研究的重点是 确定PMT中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相关的活性 激动剂卡巴胆碱进入尾外侧PMT，产生这些长期 方面的影响.我们的假设是，胆碱能激活的网站，在 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长期表达， 增强了这些长期影响。实验将测试我们的模型， 尾外侧PMT作为REM调节区，通过使用卡巴胆碱缀合的 荧光纳米球探针，我已经开发标记传入 神经元的药理学刺激的大脑。二、格局 在这些过程中，PMT中的Fos表达证明了神经元的激活 长期影响将得到检验。第三，对LTRE/LTPE的影响， PMT中Fos蛋白的抑制将通过进行 显微注射c-fos反义寡核苷酸。最后，连接 随着LDT/PPT神经元的激活，它们所包含的神经递质， c-fos、fos B、jun-B和fos-1基因时空分布 它们表达的蛋白质，以及CREB介导的基因的信号诱导 转录将使用免疫组织化学与顺行（FITC）进行研究 葡聚糖）和逆行（罗丹明纳米球）标记。成果的 实验应该为理解细胞和分子生物学提供基础。 尾外侧PMT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。的 其影响超越了睡眠，延伸到所有适应性行为状态， 神经系统必须以统一的方式反应，以重置其激活水平 以及大脑某个部分的急性突触事件如何触发改变 在基因产物中。

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.