Pontine Control of REM Atonia

项目摘要/摘要 快速眼动睡眠的一个重要特征是几乎所有肌肉的瘫痪。在REM睡眠中 行为障碍(RBD)，缺乏REM紧张症可以出现梦境 制定法律。这种疾病与神经退行性疾病密切相关，如 作为帕金森氏病，它通常出现在早期阶段，当疾病涉及 脑干的下部，包括脑桥。相当多的证据表明 脑桥背外侧下核(SLD)神经元，也称为 亚核，在快速眼动睡眠期间产生肌肉张力。这一地区的病变会产生 REM没有紧张性，而药物激活则触发类似REM的状态。我们 最近发现SLD中的谷氨酸能REM活性神经元直接投射到 脊髓内的甘氨酸能/GABA能前运动神经元。我们的假设是，这些 脊髓投射系统性红斑狼疮(SLDSP)神经元在肌肉的生成中起着关键作用 快速眼动睡眠中的紧张症。我们将研究细胞的电生理活动。 胆碱能、单胺能和食欲素能传入SLDSP神经元 在切片准备中夹住记录。我们将对SLDSP神经元进行逆行预标记 在脊髓内显微注射荧光示踪剂，然后用这些动物 对录制进行切片。在目标1和目标2中，我们将确定电生理 对胆碱能和单胺能输入的反应。我们将确定他们的预案和 突触后效应，我们将确定其受体亚型和它们的 电生理介导的机制。在目标3中，我们将重点介绍食欲素能 投入。我们发现SLDSP神经元并不直接受到食欲素的影响，但 取而代之的是，食欲素增加了对这些神经元的GABA能突触输入。我们将确定 利用诱导型增食欲素受体进行抢救实验的GABA能输入的来源 Ox1R和Ox2R敲入小鼠。这些小鼠出生时就缺乏Ox1R或Ox2R，或者两者都缺乏。我们 将含有Cre重组酶的腺相关病毒载体注射到 这些小鼠的特定脑区诱导增食欲素受体的局灶性挽救并 恢复GABA能突触传入SLDSP神经元的食欲素能调制。 总体而言，这些实验将有助于确定调控的生理基础 REM-紧张性神经元。此外，这些研究将提供对 增食欲素对REM-张力神经元的作用机制及其缺失如何导致 猝倒，以及对RBD的REM张力神经元的控制。

PROJECT SUMMARY/ABSTRACT A defining feature of REM sleep is paralysis of nearly all muscles. In REM sleep behavior disorder (RBD), the absence of REM atonia permits the appearance of dream enactment. This disorder is strongly associated with neurodegenerative diseases such as Parkinson's disease, and it often appears in early stages when the disease involves the lower brainstem including the pons. Considerable evidence demonstrates that neurons of the sublaterodorsal (SLD) nucleus of the pons, also known as the subcoreuleus, generate muscle atonia during REM sleep. Lesions of this area produce REM without atonia while pharmacological activation triggers a REM-like state. We recently found that glutamatergic REM-active neurons in the SLD directly project to the glycinergic/GABAergic premotorneurons in the spinal cord. Our hypothesis is that these spinal projecting SLD (SLDsp) neurons play a critical role in the generation of muscle atonia during REM sleep. We will examine the electrophysiological actions of the cholinergic, monoaminergic and orexinergic afferents to the SLDsp neurons using patch- clamp recordings in a slice preparation. We will retrogradely prelabel the SLDsp neurons with fluorescent tracers microinjected in the spinal cord, and then use these animals for slice recordings. In Aim 1, and Aim 2, we will determine the electrophysiological responses to cholinergic and monoaminergic inputs. We will determine their pre- and postsynaptic effects, and we will identify their receptor subtypes and their electrophysiologically-mediated mechanisms. In Aim 3, we will focus on orexinergic inputs. We have found that SLDsp neurons are not directly affected by orexin, but instead orexin increases GABAergic synaptic input to these neurons. We will identify the source of this GABAergic input with a rescue experiment using inducible orexin receptor Ox1R and Ox2R knock-in mice. These mice are born lacking Ox1R or Ox2R or both. We will place injections of an adeno-associated viral vector containing Cre recombinase into specific brain regions of these mice to induce focal rescue of orexin receptors and to restore the orexinergic modulation of GABAergic synaptic input to the SLDsp neurons. Overall these experiments will help define the physiology underlying the regulation of REM-atonia neurons. In addition, these studies will provide essential insights into the mechanism for orexin effects on REM-atonia neurons, and how their absence results in cataplexy, as well as into the control of the REM-atonia neurons in RBD.