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.

项目总结 神经系统和免疫系统之间的局部和系统相互作用在 各种生理和病理过程。这个项目的长期目标是获得一个基本的 了解神经免疫通讯如何有助于神经发育的调节， 使用一种实验模式生物黑腹果蝇的生理学和行为 进化上保守的天然免疫系统。这个应用程序的总体目标是揭示角色 以及先天免疫细胞与中枢神经系统功能相互作用的机制 癫痫易感苍蝇电压门控钠(NAV)对神经元兴奋性和行为多动的调节 海峡变种人，帕拉苏。该项目基于耐人寻味的发现：1)Parashu成人癫痫发作的严重程度 饲料中添加-3多不饱和脂肪酸(PUFA)和-亚麻酸显著抑制 (ALA)；2)血细胞(巨噬细胞样血细胞)特异性敲除哺乳动物果蝇同源基因GstS1 前列腺素D合成酶，模拟饮食ALA的影响；以及3)饮食ALA和GstS1基因敲除都是 在发育过程中有效抑制成年突变体的癫痫发作。中心假设是严重程度 成年鹦鹉的神经发育是通过生物体内的天然免疫细胞的作用来调节的。 涉及生物活性脂质介体的过程。将追求两个具体目标来调查这一假说： 1)明确血细胞在神经兴奋性和行为多动调节中的作用 突变体；以及2)确定参与成人血细胞依赖调节的基因和信号通路 癫痫发作。对于第一个目标，血细胞功能将受到遗传干扰，并对神经产生影响 发育、电生理特性和癫痫行为将在或 没有使用自由移动或拴住行为的成蝇以及分离的表型修饰物 从突变的幼虫脑中制备的原代神经元培养。对于第二个目标，候选基因方法 将用于识别与假定的脂质信号过程有关的基因，并将进行脂质组学分析 以确定在突变体中由多不饱和脂肪酸产生的脂类介体氧脂的变化。 有或没有表型修饰物。这项拟议的多学科研究预计将揭示 血细胞在NAV对神经元兴奋性和行为惊厥的饮食依赖性调节中的作用 通道突变体，并提供了对在很大程度上未被探索的脂质信号的新的遗传和分子洞察 在遗传易感神经表型的调节中发挥重要作用的通路。这个 该项目具有科学和临床意义，因为它将导致对塑料和 神经系统受免疫系统控制的交互性质，有望提供有价值的 对未来预防和治疗与异常免疫相关的神经系统疾病的策略的见解 功能。