Pontine Control of REM Atonia

项目总结/摘要 快速眼动睡眠的一个特征是几乎所有肌肉的麻痹。在REM睡眠中 行为障碍（RBD），缺乏快速眼动肌张力允许梦的出现 颁布。这种疾病与神经退行性疾病， 帕金森病，它通常出现在疾病的早期阶段， 包括脑桥在内的下脑干大量证据表明， 脑桥的背侧亚核（SLD）神经元，也称为 在快速眼动睡眠期间产生肌肉张力。这一区域的病变产生 无张力的REM，而药理学激活触发REM样状态。我们 最近发现，SLD中的突触能REM活性神经元直接投射到 脊髓中的甘氨酸能/GABA能前运动神经元。我们的假设是， 脊髓投射SLD（SLDsp）神经元在肌肉的生成中起着关键作用 快速眼动睡眠时的肌张力不足我们将研究的电生理行动的 用膜片钳技术观察了SLDsp神经元的胆碱能、单胺能和食欲素能传入。 在切片制备中夹持记录。我们将逆行预标记SLDsp神经元 在脊髓中显微注射荧光示踪剂，然后用这些动物 切片记录。在目标1和目标2中，我们将确定 对胆碱能和单胺能输入的反应。我们将确定他们的前-和 突触后效应，我们将确定它们的受体亚型和它们的 电生理介导的机制。在目标3中，我们将重点关注食欲素 输入。我们已经发现SLDsp神经元不直接受食欲素的影响， 相反，食欲素增加了对这些神经元的GABA能突触输入。我们将确定 用诱导性食欲素受体的拯救实验来说明这种GABA能输入的来源 Ox 1 R和Ox 2 R基因敲入小鼠。这些小鼠出生时缺乏Ox 1 R或Ox 2 R或两者兼而有之。我们 将注射含有Cre重组酶的腺相关病毒载体， 这些小鼠的特定脑区域以诱导食欲素受体的局灶性拯救， 恢复对SLDsp神经元的GABA能突触输入的食欲素能调节。 总的来说，这些实验将有助于确定调节的生理学基础， 快速眼动-肌张力神经元。此外，这些研究将提供必要的见解， 食欲素对REM-弛缓神经元的作用机制，以及它们的缺乏如何导致 紧张症，以及进入RBD中的REM-紧张神经元的控制。

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.