Dynamic Chemical Regulation of Voltage-gated Sodium Channels

电压门控钠通道的动态化学调节

• 批准号: 10266071
• 负责人: Stephen M Smith
• 金额: --
• 依托单位: PORTLAND VA MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-01-01 至 2023-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10266071
• 关键词: Action Potentials; Acute; Acute Pain; Affect; Agonist; Animals; Arrhythmia; Biochemical; Biochemistry; Biological Assay; Biophysical Process; Biophysics; Brain; Cannabinoids; Cell Line; Chemicals; Chemistry; Collaborations; Coupled; Cyclic AMP; Data; Disease; Dose; Drug Targeting; Electrophysiology (science); Elements; Endocannabinoids; Epilepsy; Fatty Acids; G Protein-Coupled Receptor Signaling; G-Protein-Coupled Receptors; GTP-Binding Protein alpha Subunits, Gs; GTP-Binding Proteins; Gene Deletion; Generations; Gilles de la Tourette syndrome; Glycerol; Goals; Huntington Disease; Ion Channel; Ion Channel Gating; Knowledge; Ligands; Lipids; Malignant Neoplasms; Mediating; Methods; Molecular; Muscle Cells; Muscle Spasticity; Muscle function; Nerve; Neurobiology; Neuronal Plasticity; Neurons; Neuropathy; Pain; Pain management; Paralysed; Patch-Clamp Techniques; Pathway interactions; Pattern; Peripheral Nervous System Diseases; Pharmaceutical Preparations; Pharmacology; Phosphatidylinositols; Physiological; Positioning Attribute; Protein Isoforms; Proteins; Publishing; Reagent; Regulation; Second Messenger Systems; Seizures; Signal Pathway; Signal Transduction; Slice; Sodium Channel; Spasm; System; Testing; Veterans; Wild Type Mouse; Work; anandamide; cannabinoid receptor; clinically relevant; experience; experimental study; improved; innovation; inorganic phosphate; interest; kidney cell; knock-down; live cell imaging; military veteran; mouse model; neocortical; neuronal cell body; neuronal excitability; neurotransmission; new therapeutic target; novel; novel therapeutics; patch clamp; periodic paralysis; receptor; screening; sensor; side effect; small hairpin RNA; tool; voltage

Voltage-gated sodium channels (VGSCs) are essential for action potential generation. Regulation of voltage- gated ion channel function is an important pathway by which neuronal signaling and brain function is regulated, and G-protein coupled receptors (GPCRs) form a major element of the endogenous transduction mechanisms by which this occurs. However, unlike other ion channels, VGSCs have been believed to be relatively insensitive to modulation by GPCR signaling. We have recently identified a pathway that is modulated by agents known to interact with the GPCR CB1 (cannabinoid receptor). This pathway is widespread, present in the vast majority of neocortical neurons, and strong enough to completely and reversibly block VGSC currents when maximally stimulated. This novel, dynamic signaling pathway is positioned to substantially modulate neuronal excitability and brain function. Detailed knowledge about the underlying mechanisms is crucial to understand its many effects. These preliminary findings may fundamentally change our understanding of the mechanism of action of endocannabinoids. The objectives of this proposal are to determine how endocannabinoids regulate VGSCs. We will complete this undertaking by studying VGSC function using patch- clamp methods and live cell imaging in neurons in acute neocortical brain slices, following acute isolation, and in primary cultures. We will employ mouse models. We are ideally suited to perform this project because of our preliminary data and expertise. Successful completion of these specific aims will characterize the mechanism of action of inhibition of sodium channels by this novel pathway and characterize a new mechanism by which endocannabinoids can affect neuroplasticity. Our rationale is that the identification and characterization of a novel and prevalent receptor(s) and downstream pathway will facilitate our understanding of a prevalent and potentially powerful neurobiological signaling pathway. Elucidation of the pathway will provide a detailed characterization of a new drug target that may be relevant to a wide range of diseases characterized by unbalanced excitability.

电压门控钠通道（VGSC）对于动作电位的产生是必不可少的。电压调节- 门控离子通道功能是调节神经元信号和脑功能的重要途径， 和G蛋白偶联受体（GPCR）构成了内源性转导机制的主要元件 发生这种情况。然而，与其他离子通道不同，VGSC被认为是相对独立的。 对GPCR信号转导的调节不敏感。我们最近发现了一种由 已知与GPCR CB 1（大麻素受体）相互作用的药物。这种途径广泛存在于 绝大多数的新皮层神经元，并强大到足以完全和可逆地阻断VGSC电流 最大限度地刺激。这种新的，动态的信号通路被定位为实质上调节 神经元兴奋性和大脑功能。对潜在机制的详细了解对于 了解它的许多影响。这些初步发现可能会从根本上改变我们对 内源性大麻素的作用机制。本提案的目标是确定如何 内源性大麻素调节VGSC。我们将通过使用补丁研究VGSC功能来完成这项任务- 在急性分离后，在急性新皮层脑切片中的神经元中的钳夹方法和活细胞成像，以及 在原代培养物中。我们将使用小鼠模型。我们非常适合执行这个项目，因为我们的 初步数据和专业知识。成功完成这些具体目标将是该机制的特点 通过这种新的途径抑制钠通道的作用，并表征了一种新的机制， 内源性大麻素可以影响神经可塑性。我们的基本原理是， 新的和普遍的受体和下游途径将有助于我们理解一种普遍的和 潜在的强大的神经生物学信号通路。阐明该途径将提供详细的 新的药物靶点的表征可能与广泛的疾病相关， 不平衡的兴奋性