Epigenomic Events in Development of the Locus Coeruleus Noradrenergic Neurons

蓝斑去甲肾上腺素能神经元发育中的表观基因组事件

• 批准号: 8179494
• 负责人: JAN Krzysztof BLUSZTAJN
• 金额: $ 24.53万
• 依托单位: BOSTON UNIVERSITY MEDICAL CAMPUS
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-08-01 至 2013-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8179494
• 关键词: Acetylcholine; Address; Adolescent; Adult; Age; Alzheimer' s Disease; Antibodies; Anxiety; Arousal; Atlases; Attention; Attention deficit hyperactivity disorder; Bioinformatics; Biological Assay; Brain; Brain region; Capital; Cells; Chromatin; Collection; Communities; DNA; DNA Methylation; Data; Databases; Development; Developmental Gene; Disease; Dopamine; Embryo; Epigenetic Process; Epitopes; Event; Exploratory/Developmental Grant; Female; Fetal Development; Fetus; Fluorescence-Activated Cell Sorting; Functional disorder; Funding; Funding Mechanisms; Galanin; Gene Expression; Gene Expression Profile; Generations; Genes; Goals; Histone H3; Histones; Housing; Language; Learning; Letters; Longevity; Maps; Memory; Mental Depression; Mental Health; Mental disorders; Messenger RNA; Methods; Methylation; MicroRNAs; Mixed Function Oxygenases; Modification; Molecular Profiling; Mus; Names; Nervous system structure; Neuraxis; Neurons; Neuropeptides; Neurotransmitters; Parkinson Disease; Pattern; Performance; Perinatal; Phenotype; Population; Process; Proteins; Psyche structure; Regulatory Element; Reporter; Reporting; Research; Resources; Role; Serotonin; Sleep; Sleep Disorders; Specificity; Stress; Substance P; Symptoms; Synapses; Transgenic Mice; Transgenic Organisms; United States National Institutes of Health; Universities; base; bisulfite; brain cell; chromatin immunoprecipitation; design; enhanced green fluorescent protein; epigenomics; fetal; gamma-Aminobutyric Acid; genome-wide; insight; locus ceruleus structure; male; neurodevelopment; neurogenesis; noradrenergic; novel; postnatal; response; tool; transcriptomics; young adult

DESCRIPTION (provided by applicant): This R21 application was prepared in response to the RFA entitled "Epigenomic Modifications in Neurodevelopment." As described in the RFA, our studies are broadly designed to characterize epigenetic events involved in neurodevelopmental processes and to identify epigenome-wide marks associated with vulnerability to mental disorders. The proposed studies will address cell specificity in the brain across development, and focus on how several epigenetic modifications contribute to the development and maturation of a defined population of brain cells known as the locus coeruleus noradrenergic neurons (LCNN). LCNN constitute a critical component of the circuitry that is necessary for the processes of learning, memory, attention, sleep, arousal, anxiety, and responses to stress. Abnormalities of the LCNN are thought to contribute to the symptoms of attention deficit hyperactivity disorder, depression, Parkinson's and Alzheimer's diseases, and sleep disorders. Our studies focus on the key periods of development of LCNN from the completion of neurogenesis in the fetus through postnatal maturation and the establishment of functional synaptic connections, and conclude with adulthood in both male and female subjects. We have developed a method to purify defined neuronal populations throughout lifespan by fluorescence-activated cell sorting using transgenic mice that express enhanced green fluorescent protein (EGFP) under the control of the regulatory elements of genes expressed exclusively in these populations. In this study we will use transgenic mice (designated Dbh- EGFP) that express EGFP under the control of the regulatory elements of the dopamine 2-hydroxylase (Dbh) gene - a marker of noradrenergic neurons. By combining several state-of-the-art experimental approaches, we will generate the first comprehensive characterization of the genome-wide developmental pattern of the LCNN transcriptome [for both messenger RNA (mRNA) and microRNA (miRNA)], the DNA CpG methylome, and genome-wide maps of methylated histone H3 (e.g. H3K4me1, H3K4me2, H3K4me3, H3K9me3, H3K27me3). We will integrate this novel information into a framework defined by chromatin state maps and developmental gene expression profiles of LCNN. We will develop interactive bioinformatics tools designed to manage, visualize and integrate these data. This study, supported by an R21 mechanism that focuses on a single population of neurons highly relevant to mental health (the LCNN), will serve as a proof of concept for our long- term goal: to establish a comprehensive atlas of developmental epigenomic events that occur in key neuronal populations purified from specific brain regions and identified by their neurotransmitter (e.g. GABA, dopamine, serotonin, acetylcholine) or neuropeptide (e.g. galanin, NPY, VIP, substance P) phenotype. A large collection of the relevant EGFP-expressing transgenic mouse lines has been generated and is available at the NIH- supported GENSAT resource. PUBLIC HEALTH RELEVANCE: Accumulating evidence indicates that epigenetic mechanisms are vital for normal brain development and function in the adult, and suggests that epigenetic abnormalities may contribute to the pathophysiology of mental illness. We will study the role of genome-wide epigenetic events in the development of locus coeruleus noradrenergic neurons, which are necessary for the processes of learning, memory, attention, sleep, arousal, anxiety, and responses to stress. Abnormalities in the function of these neurons are thought to contribute to the symptoms of attention deficit hyperactivity disorder, depression, Parkinson's and Alzheimer's diseases, and sleep disorders.

描述（由申请人提供）：此 R21 申请是为了响应题为“神经发育中的表观基因组修饰”的 RFA 而准备的。正如 RFA 中所述，我们的研究广泛旨在描述神经发育过程中涉及的表观遗传事件，并识别与精神障碍易感性相关的表观基因组范围内的标记。拟议的研究将解决大脑在整个发育过程中的细胞特异性，并重点关注几种表观遗传修饰如何促进被称为蓝斑去甲肾上腺素神经元（LCNN）的特定脑细胞群的发育和成熟。 LCNN 构成了学习、记忆、注意力、睡眠、唤醒、焦虑和压力反应过程所必需的电路的关键组成部分。 LCNN 的异常被认为会导致注意力缺陷多动障碍、抑郁症、帕金森病、阿尔茨海默病以及睡眠障碍的症状。我们的研究重点关注男性和女性受试者的 LCNN 发育的关键时期，从胎儿神经发生的完成到出生后的成熟和功能性突触连接的建立，直至成年。我们开发了一种方法，通过使用转基因小鼠进行荧光激活细胞分选来纯化整个生命周期中确定的神经元群体，这些转基因小鼠在这些群体中专门表达的基因的调控元件的控制下表达增强型绿色荧光蛋白（EGFP）。在这项研究中，我们将使用转基因小鼠（指定为 Dbh-EGFP），它们在多巴胺 2-羟化酶 (Dbh) 基因（去甲肾上腺素能神经元的标记）的调节元件的控制下表达 EGFP。通过结合几种最先进的实验方法，我们将首次全面表征 LCNN 转录组（信使 RNA (mRNA) 和 microRNA (miRNA)）的全基因组发育模式，即 DNA CpG 甲基化组，以及甲基化组蛋白 H3 的全基因组图谱（例如 H3K4me1、H3K4me2、H3K4me3、H3K9me3、H3K27me3）。我们将把这些新信息整合到由染色质状态图和 LCNN 发育基因表达谱定义的框架中。我们将开发交互式生物信息学工具，旨在管理、可视化和整合这些数据。这项研究得到了 R21 机制的支持，该机制专注于与心理健康高度相关的单个神经元群体（LCNN），将作为我们长期目标的概念证明：建立一个全面的发育表观基因组事件图谱，发生在从特定大脑区域纯化的关键神经元群体中，并通过其神经递质（例如 GABA、多巴胺、血清素、乙酰胆碱）或神经肽（例如甘丙肽、 NPY、VIP、P 物质）表型。已经生成了大量相关的表达 EGFP 的转基因小鼠品系，并且可以在 NIH 支持的 GENSAT 资源中获取。 公共卫生相关性：越来越多的证据表明表观遗传机制对于成人的正常大脑发育和功能至关重要，并表明表观遗传异常可能导致精神疾病的病理生理学。我们将研究全基因组表观遗传事件在蓝斑去甲肾上腺素能神经元发育中的作用，这些神经元对于学习、记忆、注意力、睡眠、觉醒、焦虑和压力反应过程是必需的。这些神经元功能异常被认为会导致注意力缺陷多动障碍、抑郁症、帕金森病和阿尔茨海默病以及睡眠障碍的症状。