The neural circuitry of seizure-induced apnea and SUDEP

癫痫发作引起的呼吸暂停和 SUDEP 的神经回路

• 批准号: 10719519
• 负责人: Ian Christopher Wenker
• 金额: $ 40.38万
• 依托单位: UNIVERSITY OF VIRGINIA
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-06-01 至 2028-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10719519
• 关键词: Acute; Address; Apnea; Benchmarking; Brain; Brain Mapping; Brain Stem; Breathing; Bypass; Cardiac; Cause of Death; Cell Nucleus; Cessation of life; Chromosome Mapping; Clonic Seizure; Data; Development; Electrophysiology (science); Elements; Epilepsy; Experimental Designs; FOXP2 gene; Failure; Generations; Genetic; Heart Arrest; In Vitro; Individual; Injections; Interruption; Maps; Midbrain structure; Modeling; Motor Seizures; Mus; Muscle; Mutant Strains Mice; Mutation; National Institute of Neurological Disorders and Stroke; Nerve Fibers; Neurons; Nodal; Patients; Pattern; Phase; Pontine structure; Prevention strategy; Prosencephalon; Recovery; Refractory; Research; Respiratory Failure; Risk Factors; Role; SCN8A gene; Seizures; Site; Slice; Structure; Sudden Death; Techniques; Testing; Tonic Seizures; Vagus nerve structure; Work; afferent nerve; hindbrain; in vivo; mouse model; mutant; neural; neural circuit; neuronal circuitry; neuronal excitability; novel; optogenetics; parabrachial nucleus; patch clamp; prevent; receptor; recruit; respiratory; selective expression; sudden unexpected death in epilepsy; ventilation

Sudden Unexpected Death in Epilepsy (SUDEP) accounts for up to 17% of all epilepsy-related deaths, and 50% for those refractory to treatment. There is increasing evidence that apnea (respiratory arrest) is the primary cause of death from SUDEP. We have shown in a number of mouse models - including PTZ injection, rapid kindling (prelim. data), and mouse models harboring Scn8a mutations identified from SUDEP patients - that sudden death is due to seizure-induced apnea (SIA) that occurs during tonic seizures, minutes prior to terminal asystole. As it pertains to this proposal, we have also made four other important observations: 1) artificial ventilation can prevent death and suppressed cardiac activity does not increase likelihood of fatality; 2) tonic contraction of breathing muscles represents a likely mechanism of SIA; 3) inhibition of cortical seizure activity does not impact SIA; and 4) inhibition of the inspiratory oscillator does not reduce apnea (prelim. data), suggesting seizure spread bypasses homeostatic elements of respiratory control circuitry. CENTRAL HYPOTHESIS: [AIM 1] Increased midbrain and pontine excitability is both necessary and sufficient to produce SIA, [AIM 2] the parabrachial/kölliker fuse (PB/KF) nucleus in the pons represents a key nodal point for SIA generation, and [AIM 3] that brainstem nuclei, like the PB/KF and PAG are recruited specifically during SIA, which we will explore using electrophysiology and genetic mapping. AIM 1: We have previously used chemogenetics to demonstrate that cortical ictal activity is not required for SIA and we now propose that hyperexcitability of the pons and midbrain drive SIA and SUDEP. We will use chemogenetics to inhibit different parts of the brainstem of Scn8a mutant mice to test their necessity for SIA and SUDEP. Additionally, we will test sufficiency of these regions by selectively expressing an Scn8a mutation that increases neuronal excitability. Using the rapid kindling model, we will also perform patch clamp recordings to test whether increased neuronal excitability in these regions occurs concurrently with the development of SIA. AIM 2: The parabrachial/kölliker fuse nucleus PB/KF in the Pons is a key nodal point for descending forebrain input and projects past the inspiratory oscillator, and is modulated by the vagus nerve. In this aim, we will photoinhibit the Foxp2-positive PB/KF neurons to regain normal breathing pattern during SIA, photostimulate P2ry1-positive vagal afferent nerve fibers to interrupt inspiration during SIA, and determine if PB/KF and Npy2r-positive vagal neuron photostimulation rescues postictal breathing in our PTZ SUDEP mouse model. AIM 3: We have demonstrated that tonic seizures with SIA and clonic seizures without SIA can occur in the very same mouse, both in Scn8a mutant and rapid kindling models (prelim. data) and that certain mid- and hindbrain structures are activated during SIA. We will determine the neuronal circuitry activated specifically during SIA and SUDEP using in vivo electrophysiology and TRAP2 identification of activated neurons.

癫痫猝死 (SUDEP) 占所有癫痫相关死亡的 17%，并且 50% 的患者难以接受治疗。越来越多的证据表明，呼吸暂停（呼吸骤停）是导致这种情况的原因。 我们已经在许多小鼠模型中证明了 SUDEP 死亡的主要原因 - 包括 PTZ 注射， 快速点燃（初步数据），以及从 SUDEP 患者中鉴定出的携带 Scn8a 突变的小鼠模型 - 猝死是由于癫痫发作引起的呼吸暂停 (SIA)，发生在强直性癫痫发作前几分钟 就终止心脏停搏而言，我们还提出了其他四项重要观察结果： 1) 人工通气可以预防死亡，抑制心脏活动不会增加死亡的可能性； 2) 呼吸肌的强直收缩是 SIA 的一个可能机制 3) 抑制皮质癫痫； 活动不影响 SIA；4) 抑制吸气振荡器不会减少呼吸暂停（初步数据）， 表明癫痫发作绕过了呼吸控制回路的稳态元件。 假设：[目标 1] 中脑和脑桥兴奋性的增加对于 产生 SIA，[AIM 2] 脑桥中的臂旁/kölliker 融合 (PB/KF) 核代表关键节点 SIA 生成点，以及招募脑干核团（如 PB/KF 和 PAG）的 [AIM 3] 特别是在 SIA 期间，我们将使用电生理学和基因图谱进行探索：我们。 之前已经使用化学遗传学证明 SIA 不需要皮质发作活动，现在我们 提出脑桥和中脑的过度兴奋驱动 SIA 和 SUDEP 我们将使用化学遗传学来研究。 抑制Scn8a突变小鼠脑干的不同部位，以测试其对SIA和SUDEP的必要性。 此外，我们将通过选择性表达 Scn8a 突变来测试这些区域的充分性，该突变会增加 使用快速点燃模型，我们还将进行膜片钳记录来测试是否。 这些区域的神经兴奋性增加与 SIA 2 的发展同时发生： 脑桥中的臂旁/科利克融合核 PB/KF 是前脑下降输入和 投射通过吸气振荡器，并由迷走神经调节。为此，我们将光抑制。 Foxp2 阳性 PB/KF 神经元在 SIA 期间恢复正常呼吸模式，光刺激 P2ry1 阳性 迷走神经传入神经纤维在 SIA 期间中断吸气，并确定 PB/KF 和 Npy2r 阳性迷走神经是否 神经元光刺激可挽救我们的 PTZ SUDEP AIM 3 小鼠模型的发作后呼吸。 证明伴有 SIA 的强直性癫痫发作和不伴有 SIA 的阵挛性癫痫发作可以发生在同一只小鼠身上， 在 Scn8a 突变体和快速点燃模型（初步数据）中，某些中脑和后脑结构是 我们将使用 SIA 和 SUDEP 期间特别激活的神经回路来确定。 激活神经元的体内电生理学和 TRAP2 鉴定。