Roles of hemocytes and bioactive lipids in the modulation of neuronal excitability and seizure behavior in Drosophila voltage-gated sodium channel mutants

血细胞和生物活性脂质在果蝇电压门控钠通道突变体神经元兴奋性和癫痫行为调节中的作用

• 批准号: 10433305
• 负责人: TOSHIHIRO KITAMOTO
• 金额: $ 23.18万
• 依托单位: UNIVERSITY OF IOWA
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-02-15 至 2024-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10433305
• 关键词: Adult; Affect; Animal Model; Basic Science; Behavior; Behavior Control; Behavioral; Biological; Biological Process; Biology; Blood Cells; Brain; Candidate Disease Gene; Cells; Communication; Complex; DNA Sequence Alteration; Development; Diet; Dietary Component; Drosophila genus; Drosophila melanogaster; Electrophysiology (science); Experimental Models; Family; Foundations; Future; Genes; Genetic; Genetic Predisposition to Disease; Genetic Screening; Glutathione S-Transferase; Goals; Hematopoietic; Hemocytes; Hyperactivity; Immune; Immune response; Immune system; Innate Immune Response; Innate Immune System; Insecta; Interdisciplinary Study; Larva; Link; Linolenic Acids; Lipids; Mediating; Mental disorders; Molecular; Morphology; Natural Immunity; Nature; Nervous system structure; Neuraxis; Neuroimmune; Neurologic; Neurons; Neurophysiology - biologic function; Orthologous Gene; Outcome; Pathologic Processes; Phenotype; Physiological; Physiological Processes; Physiology; Play; Polyunsaturated Fatty Acids; Process; Property; Prostaglandins; Protocols documentation; Public Health; Pupa; Recipe; Research; Role; Seizures; Severities; Signal Pathway; Signal Transduction; Sodium; Sodium Channel; Solid; Synthase D; base; behavioral phenotyping; clinically significant; dietary; feeding; fly; food standard; genetic manipulation; immune function; insight; knock-down; lipid mediator; lipidomics; macrophage; mutant; nervous system disorder; neurodevelopment; neuronal excitability; neuropsychiatric disorder; neuroregulation; novel; novel strategies; prevent; voltage

PROJECT SUMMARY Local and systemic interactions between the nervous system and immune system play important roles in various physiological and pathological processes. The long-term goal of this project is to obtain a fundamental understanding of how neuroimmune communications contribute to the regulation of neural development, physiology, and behavior using an experimental model organism Drosophila melanogaster possessing the evolutionarily conserved innate immune system. The overall objective in this application is to reveal the roles and mechanisms of functional interactions between innate immune cells and the central nervous system in the modulation of neuronal excitability and behavioral hyperactivity in seizure-prone fly voltage-gated sodium (Nav) channel mutant, paraShu. The project is based on intriguing findings that: 1) paraShu adult seizure severity is significantly suppressed by diet supplemented with -3 polyunsaturated fatty acids (PUFAs) -linolenic acid (ALA); 2) hemocyte (macrophage-like blood cell)-specific knockdown of GstS1, a fly ortholog of mammalian prostaglandin D synthase, mimics the effect of dietary ALA; and 3) both dietary ALA and GstS1 knockdown are effective during development to suppress seizures in adult mutants. The central hypothesis is that the severity of adult paraShu is modulated through neural development by the action of innate immune cells in the biological processes involving bioactive lipid mediators. Two specific aims will be pursued to investigate the hypothesis: 1) Define the roles of hemocytes in modulation of neuronal excitability and behavioral hyperactivity in paraShu mutants; and 2) Identify genes and signaling pathways involved in hemocyte-dependent modulation of adult seizures. For the first aim, hemocyte functions will be genetically perturbed and the effects on neural development, electrophysiological properties, and seizure behavior will be examined in the presence or absence of phenotypic modifiers using freely moving or tethered behaving adult flies as well as dissociated primary neuronal cultures prepared from mutant larval brains. For the second aim, a candidate gene approach will be used to identify genes involved in putative lipid signaling processes, and lipidomics analysis will be carried out to determine the changes in oxylipins, lipid mediators produced from PUFAs, in the mutants in the presence or absence of phenotypic modifiers. The proposed multidisciplinary research is expected to reveal the roles of hemocytes in the diet-dependent modulation of neuronal excitability and behavioral seizures in Nav channel mutants, and provide novel genetic and molecular insights into the largely unexplored lipid signaling pathways that play important roles in the modulation of genetically predisposed neurological phenotypes. The project is scientifically and clinically significant because it will lead to a deeper appreciation of the plastic and interactive nature of the nervous system controlled by the immune system, and is expected to provide valuable insights into future strategies to prevent and treat nervous system disorders associated with aberrant immune function.

项目摘要 神经系统和免疫系统之间的局部和系统性相互作用在 各种物理和病理过程。该项目的长期目标是获得基本 了解神经免疫传播如何促进神经发展的调节， 生理学和使用实验模型的有机体果蝇的行为具有 进化配置的先天免疫系统。该应用程序的总体目的是揭示角色 与先天免疫细胞与中枢神经系统之间功能相互作用的机制 调节神经元令人兴奋和行为多动症中癫痫发作电压门控钠（NAV）的调节 频道突变体，帕拉修。该项目基于有趣的发现：1） 被补充的-3多不饱和脂肪酸（PUFAS）丁醇酸的饮食明显抑制 （翼）; 2）血细胞（巨噬细胞样血细胞） - GSTS1的特异性敲低，哺乳动物的蝇直源 前列腺素D合酶，模仿饮食ALA的作用； 3）饮食中的ALA和GSTS1敲低均为 在发育过程中有效抑制成人突变体的癫痫发作。中心假设是严重性 成人帕拉修（Parashu 过程涉及生物活性脂质介质。将追求两个具体的目标来研究以下假设： 1）定义血细胞在parashu中神经元激动和行为多动症调节中的作用 突变体； 2）确定成年血细胞依赖性调节涉及的基因和信号通路 癫痫发作。为了第一个目标，血细胞功能通常会受到干扰，并且对中性的影响 在存在或 使用自由移动或束缚的成年蝇以及分离的表型修饰符没有表型修饰符 由突变幼虫大脑制备的原发性神经元培养物。对于第二个目标，候选基因方法 将用于鉴定参与推定脂质信号传导过程的基因，脂质组学分析将是 执行以确定黄脂素，PUFAS产生的脂质介质的变化，在突变体中产生的变化 存在或不存在表型修饰符。拟议的多学科研究有望揭示 血细胞在NAV中神经元令人兴奋和行为癫痫发作的饮食依赖性调节中的作用 通道突变体，并向很大程度上出乎意料的脂质信号传导提供新的遗传和分子见解 在一般易感的神经表型调节中起重要作用的途径。 项目在科学和临床上具有重要意义，因为它将对塑料和 由免疫系统控制的神经系统的互动性质，有望提供有价值的 对预防和治疗与异常免疫相关的未来策略的见解 功能。