Monoamine-Mediated Arousal to Prevent Seizure-Induced Sudden Death

项目摘要/摘要 癫痫猝死(SUDEP)是全身性致死的主要原因。 癫痫发作，目前还没有有效的疗法来治疗这种毁灭性的疾病。临床观察和临床观察 动物研究表明，癫痫诱导的呼吸骤停(S-IRA)是导致SUDEP的主要事件。 尽管目前的研究表明5-羟色胺(5-羟色胺)(也称为腺苷)与S-IRA有关，但尚不清楚是否有其他 神经递质对S-IRA的作用，以及这些神经递质如何调节S-IRA。这些差距存在于 知识在很大程度上阻碍了制定针对SUDEP的有效预防战略。 大脑中的单胺类神经递质，如5-羟色胺、去甲肾上腺素(NE)和多巴胺(DA)， 促进唤醒，并可能刺激呼吸。在我们对DBA/1小鼠的初步和已发表的研究中 模型后，我们确定了5-羟色胺或去甲肾上腺素水平的药理学升高，以及5-羟色胺的光遗传刺激。 中介唤醒抑制S-IRA。因此，在这个高风险和高影响的提案中，我们假设 增强的单胺介导的唤醒可防止癫痫诱发的小鼠猝死。我们将测试 这一假设在两个不同的SUDEP小鼠模型中，DBA/1小鼠和DraveT小鼠表现出 人类德拉韦综合征(癫痫的一种)的显著特征，包括自发性癫痫发作。在……里面 具体目标1，我们将确定单胺功能的药物增强是否会降低S-IRA和/或 DBA/1和Dravet小鼠均发生猝死。我们将检查5-羟色胺、去甲肾上腺素和多巴胺是否与癫痫有关- 使用单胺再摄取抑制剂的相关死亡率，并确定特定的5-羟色胺能、肾上腺素能和 参与预防癫痫相关死亡的多巴胺能受体。在具体目标2中，我们将 确定选择性刺激中缝背侧、蓝斑或腹侧被盖区是否降低S-IRA 在DBA/1和Dravet两种小鼠中。我们将使用光遗传学来激活大脑中的单胺类神经核团 已知可调节唤醒反应，并检查是否由每种单胺介导的增强唤醒 打压S-艾拉。如果成功，拟议的研究可能导致在概念上的重大进展 SUDEP的机制，并促进新的药理和神经刺激策略的开发 对苏德普的指控。拟议研究的完成将建立动物模型和方法学，以便进一步 解剖神经元电路，研究相互作用，并阐明细胞机制 与癫痫相关死亡有关的单胺类物质。

PROJECT SUMMARY/ABSTRACT Sudden unexpected death in epilepsy (SUDEP) is the major cause of deaths associated with generalized seizures, and there are currently no effective therapies for this devastating disorder. Clinical observations and animal studies suggest that seizure-induced respiratory arrest (S-IRA) is the primary event leading to SUDEP. Although current studies implicate serotonin (5-HT) (also adenosine) in S-IRA, it is unclear if other neurotransmitters contribute to S-IRA, and how these neurotransmitters modulate S-IRA. These gaps in knowledge have significantly hindered the development of effective prevention strategies against SUDEP. Monoamine neurotransmitters in the brain, such as 5-HT, norepinephrine (NE) and dopamine (DA), are arousal-promoting and may stimulate breathing. In our preliminary and published studies in the DBA/1 mouse model, we determined that pharmacologic elevation of 5-HT or NE levels, and optogenetic stimulation of 5-HT- mediated arousal suppress S-IRA. Thus, in this high risk and high impact proposal, we hypothesize that enhanced monoamine-mediated arousal prevents seizure-induced sudden death in mice. We will test this hypothesis in two separate SUDEP mouse models, the DBA/1 mouse and the Dravet mouse that exhibits the salient features of Dravet syndrome (a type of epilepsy) in humans, including spontaneous seizures. In specific aim 1, we will determine if pharmacologic enhancement of monoamine function reduces S-IRA and/or sudden death in both DBA/1 and Dravet mice. We will examine if 5-HT, NE and DA are implicated in seizure- related mortality using monoamine reuptake inhibitors, and identify the specific serotonergic, adrenergic and dopaminergic receptors that are involved in prevention of seizure-related mortality. In specific aim 2, we will determine if selective stimulation of the dorsal raphe, locus coeruleus or ventral tegmental area reduces S-IRA in both DBA/1 and Dravet mice. We will use optogenetics to activate the monoaminergic brain nuclei that are known to mediate arousal response, and examine if enhancing arousal mediated by each monoamine suppresses S-IRA. If successful, the proposed studies could lead to significant conceptual advance in the mechanisms of SUDEP, and foster development of novel pharmacologic and neurostimulatory strategies against SUDEP. Completion of the proposed studies will establish animal models and methodology for further dissecting the neuronal circuitry, investigating the interaction, and elucidating the cellular mechanisms of monoamines involved in seizure-related mortality.