Sodium and Calcium Channels: Structure, Function, Neuroplasticity, and Disease

钠和钙通道：结构、功能、神经可塑性和疾病

• 批准号: 9923774
• 负责人: WILLIAM A CATTERALL
• 金额: $ 111.16万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-05-01 至 2027-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/9923774
• 关键词: Action Potentials; Aging; Ataxia; Brain; Calcium; Calcium Channel; Calmodulin; Cessation of life; Characteristics; Childhood; Circadian Rhythm Sleep Disorders; Cognitive deficits; Combined Modality Therapy; Communication; Complex; Cre-LoxP; Disease; Elements; Epilepsy; Erythromelalgia; Failure; Functional disorder; Genetic Models; Genetic study; Hippocampus (Brain); Impairment; Inherited; Interneurons; Ion Channel; Ions; Learning; Memory; Methods; Migraine; Modification; Molecular; Molecular Structure; Mus; Mutation; Neurodegenerative Disorders; Neuronal Plasticity; Neurons; Paroxysmal extreme pain disorder; Physiological; Process; Property; Proteins; Regulation; Resolution; Role; Seizures; Signal Transduction; Signaling Protein; Site; Sodium; Sodium Channel; Status Epilepticus; Structure; Synapses; Synaptic Transmission; Synaptic plasticity; Syndrome; Work; X-Ray Crystallography; autistic; base; chronic pain; dravet syndrome; excitatory neuron; experimental study; in vivo; insight; loss of function mutation; molecular drug target; mouse genetics; mouse model; neural circuit; neuropsychiatric disorder; neurotransmitter release; next generation; novel; novel strategies; periodic paralysis; premature; presynaptic; sensor; sudden unexpected death in epilepsy; three dimensional structure; voltage

Voltage-gated sodium (Nav) and calcium (Cav) channels generate action potentials and initiate synaptic transmission in neurons. Mutations in them cause inherited epilepsy, migraine, chronic pain, and periodic paralysis, and they are important molecular targets for drugs. A. New insights into structure and function of Nav channels have come from our high-resolution x-ray crystallography of their bacterial ancestor NavAb. We will further define the structural basis for key functional properties of mammalian Nav channels by building their characteristic structural features into NavAb, including the structural basis for voltage-dependent activation, ion selectivity, and fast inactivation. Based on these results, we will determine the structural basis for impaired Nav channel function by mutations that cause periodic paralysis and the chronic pain syndromes erythromelalgia and paroxysmal extreme pain disorder. B. Failure of learning and memory is a debilitating aspect of aging and neurodegenerative disease, yet we do not understand the basic mechanisms of these crucial brain processes and we cannot intervene effectively in these deficits. Learning and memory takes place primarily at synapses. Presynaptic calcium (Cav2.1) channels initiate neurotransmitter release at most synapses in the brain. The activity of these channels is tightly regulated by a large complex of signaling proteins, including calmodulin and related calcium-sensor proteins. Our work implicates Cav2.1 channel regulation in short-term synaptic plasticity in transfected synapses in cultured neurons and in a novel mouse model in which the IM-AA mutation is inserted into Cav2.1. We will further define the molecular and structural mechanism for Cav2.1 channel regulation, determine the role of regulation of Cav2.1 channels in short-term synaptic plasticity of neural circuits, and explore the role of regulation of Cav2.1 channels and short-term synaptic plasticity in spatial learning and memory. Our experiments with this unique mouse model will give unique insights into the mechanism of short-term presynaptic plasticity in hippocampal neurons and its role in integrative bbrain function. C. Dravet Syndrome (DS) is a devastating childhood neuropsychiatric disorder caused by de novo, heterozygous loss-of-function mutations in Nav1.1. We developed a mouse genetic model with all the features of DS, including thermally induced and spontaneous seizures, ataxia, circadian rhythm and sleep disorders, cognitive deficit, autistic-like features, and premature death via SUDEP. Physiological and genetic studies show that all these effects are correlated with loss of Na currents and excitability of GABAergic interneurons, without consistent effects on excitatory neurons, which causes imbalance of excitation vs. inhibition in neural circuits. To further advance understanding of pathophysiology and treatment of DS, we will determine the neural cells and circuits responsible for DS using specific deletion by the Cre-Lox method, identify the sites of hyperexcitability in neural cells and circuits that appear first in DS mice in vivo, and optimize next-generation combination therapy for seizures, status epilepticus, cognitive deficit, and premature death in DS.

电压门控钠（NAV）和钙（CAV）通道产生动作电位并启动突触 神经元的传播。其中的突变导致遗传癫痫，偏头痛，慢性疼痛和周期性 瘫痪，它们是药物的重要分子靶标。答：对导航的结构和功能的新见解 通道来自我们的细菌祖先Navab的高分辨率X射线晶体学。我们将 进一步定义哺乳动物NAV通道的关键功能特性的结构基础 NAVAB中的特征结构特征，包括电压依赖性激活的结构基础，离子 选择性和快速失活。基于这些结果，我们将确定NAV受损的结构基础 通道功能通过引起周期性麻痹和慢性疼痛综合症的突变功能 和阵发性极端疼痛障碍。 B.学习和记忆的失败是衰老和 神经退行性疾病，但我们不了解这些关键大脑过程的基本机制 而且我们无法有效干预这些缺陷。学习和记忆主要发生在突触。 突触前钙（CAV2.1）通道启动大脑中大多数突触的神经递质释放。这 这些通道的活性受到大量信号蛋白的严格调节，包括钙调蛋白和 相关的钙传感器蛋白。我们的工作暗示CAV2.1在短期突触可塑性中调节 在培养的神经元中转染的突触和IM-AA突变的新型小鼠模型中 插入Cav2.1。我们将进一步定义CAV2.1通道的分子和结构机制 调节，确定CAV2.1通道调节在神经短期突触可塑性中的作用 电路，并探讨Cav2.1通道调节和短期突触可塑性在空间中的作用 学习和记忆。我们对这种独特鼠标模型的实验将为您提供独特的见解 海马神经元中短期突触前可塑性的机制及其在综合BBRAIN中的作用 功能。 C. dravet综合征（DS）是由从头开始引起的毁灭性儿童神经精神疾病 NAV1.1中的杂合丧失功能突变。我们开发了一个具有所有特征的鼠标遗传模型 DS，包括热诱导和自发性癫痫发作，共济失调，昼夜节律和睡眠障碍， 认知缺陷，自闭症特征和Sudep过早死亡。生理学研究 表明所有这些效果与Na电流的丧失和GABA能中间神经元的兴奋性相关， 对兴奋性神经元没有一致的影响，这会导致激发不平衡与神经的抑制作用 电路。为了进一步了解病理生理学和DS的治疗，我们将确定 使用Cre-Lox方法使用特定缺失负责DS的神经细胞和电路，确定 在体内DS小鼠中首先出现的神经细胞和电路中的过度兴奋性，并优化了下一代 DS的癫痫发作，癫痫持续状态，认知缺陷和过早死亡的组合疗法。