Epigenetic programming of brain development by choline nutrition

胆碱营养对大脑发育的表观遗传编程

基本信息

批准号：
7992008
负责人：
JAN Krzysztof BLUSZTAJN
金额：
$ 24.38万
依托单位：
BOSTON UNIVERSITY MEDICAL CAMPUS
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-09-20 至 2012-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7992008
关键词：
Acetylcholine Acetylcholinesterase Adolescent Adult Age Antibodies Area Arts Attention Basal Nucleus of Meynert Brain Capital Cerebral cortex Choline Choline O-Acetyltransferase Chromatin DNA DNA Methylation Data Development Developmental Gene Diet Dietary intake Embryo Epigenetic Process Epitopes Event Fetal Development Fetus Fluorescence-Activated Cell Sorting Gene Expression Gene Expression Profile Generations Genes Hippocampus (Brain)Histone H3 Histones Human Development Immunoprecipitation Intake Investigation Knowledge Lactation Language Learning Letters Longevity Lysine Maps Measures Medial Mediating Memory Messenger RNA Metabolic Methods Methylation Methyltransferase MicroRNAs Microarray Analysis Mission Modeling Modification Molecular Profiling Mothers Mus Names National Institute of Child Health and Human Development Neuroglia Neurons Neurotransmitters Nutrient Nutritional Pattern Perinatal Phenotype Population Pregnancy Process Proteins Regulatory Element Resources S-Adenosylmethionine Sleep Staging Synapses Testing Time Transgenic Mice Weaning basal forebrain cholinergic neurons body system brain cell cell type choline transporter cholinergic chromatin immunoprecipitation critical period design developmental neurobiology dietary supplements enhanced green fluorescent protein enzyme activity feeding fetal genome-wide histone methyltransferase methyl group myelination neurogenesis neuron development neuronal cell body neuronal circuitry neurotransmission novel nutrition offspring postnatal prenatal programs public health relevance response synaptogenesis young adult

项目摘要

DESCRIPTION (provided by applicant): Accumulating evidence indicates that epigenetic mechanisms are vital for normal brain development including modulation of generation of neurons and glia, neuronal cell-type specification and differentiation, circuit and synapse formation, and myelination. These events - that occur during a critical period that spans gestation and early postnatal time - are highly sensitive to the supply of essential nutrients and in particular to the supply of metabolic methyl group donors including choline. The central hypothesis to be tested by the studies described in this proposal is that alterations in the availability of choline during this critical period modulates the differentiation and phenotype of basal forebrain cholinergic neurons (BFCN) via epigenetic mechanisms involving changes in DNA and histone methylation. BFCN constitute a key component of the neuronal circuitry that is necessary for the processes of learning, memory, attention, and sleep. We have developed a method to purify BFCN throughout lifespan by fluorescence-activated cell sorting using transgenic mice that that express enhanced green fluorescent protein (eGFP) under the control of the regulatory elements of the choline acetyltransferase gene. Using purified BFCN from mice at various developmental stages between embryonic day 16 and postnatal day 30 we will determine: 1) global transcriptome profile using microarrays; 2) DNA methylation patterns with methylated DNA immunoprecipitation (MeDIP) followed by microarray analysis and 3) chromatin immunoprecipitation (ChIP) with antibodies against different methylated epitopes on specific lysine residues of histone 3 (e.g. H3K9me2; H3K4me1; H3K4me3) followed microarray analysis (ChIP-chip). These data will constitute the first characterization of the epigenome and its correlation with the transcriptome of a developing pure neuronal population. We will perform an analogous study on the fetuses and offspring of mice whose mothers consume diets containing choline-deficient, choline-sufficient, or choline-supplemented diets throughout pregnancy until weaning. The results of the investigations will provide information on epigenetic processes that are associated with the completion of developmental milestones of BFCN and on the modulation of these processes by gestation and postnatal nutritional choline intake. PUBLIC HEALTH RELEVANCE: The basal forebrain cholinergic neurons (BFCN) constitute a key component of the neuronal circuitry that is necessary for the processes of learning, memory, attention, and sleep. BFCN development is modulated by the perinatal availability of the methyl-group-donating essential nutrient, choline, and changes of choline intake in pregnancy cause dramatic modifications in DNA and histone methylation patterns in fetal brain. We will perform the first of its kind study on murine BFCN purified by FACS throughout development (fetal to adulthood) by measuring the effects of choline intake on the pattern of BFCN gene expression in correlation with global DNA and histone methylation patterns.

描述（由申请人提供）：越来越多的证据表明，表观遗传机制对于正常的大脑发育至关重要，包括调节神经元和神经胶质的生成、神经元细胞类型的规范和分化、回路和突触的形成以及髓鞘形成。这些事件发生在妊娠期和产后早期的关键时期，对必需营养素的供应，特别是对包括胆碱在内的代谢甲基供体的供应高度敏感。本提案中描述的研究要测试的中心假设是，在此关键时期胆碱可用性的变化通过涉及 DNA 和组蛋白甲基化变化的表观遗传机制调节基底前脑胆碱能神经元 (BFCN) 的分化和表型。 BFCN 是学习、记忆、注意力和睡眠过程所必需的神经元回路的关键组成部分。我们开发了一种在整个生命周期中通过荧光激活细胞分选来纯化 BFCN 的方法，使用转基因小鼠进行荧光激活细胞分选，这些小鼠在胆碱乙酰转移酶基因的调控元件的控制下表达增强型绿色荧光蛋白 (eGFP)。使用来自胚胎第 16 天和出生后第 30 天之间不同发育阶段的小鼠的纯化 BFCN，我们将确定：1) 使用微阵列的整体转录组谱； 2) 使用甲基化 DNA 免疫沉淀 (MeDIP) 进行 DNA 甲基化模式分析，然后进行微阵列分析；3) 使用抗组蛋白 3 特定赖氨酸残基上不同甲基化表位的抗体进行染色质免疫沉淀 (ChIP)（例如 H3K9me2；H3K4me1；H3K4me3），然后进行微阵列分析 (ChIP) -芯片）。这些数据将构成表观基因组的第一个特征及其与发育中的纯神经元群体转录组的相关性。我们将对小鼠的胎儿和后代进行类似的研究，这些小鼠的母亲在整个怀孕期间直到断奶期间摄入胆碱缺乏、胆碱充足或补充胆碱的饮食。研究结果将提供与 BFCN 发育里程碑完成相关的表观遗传过程以及妊娠和产后营养胆碱摄入对这些过程的调节信息。公共健康相关性：基底前脑胆碱能神经元 (BFCN) 是学习、记忆、注意力和睡眠过程所必需的神经元回路的关键组成部分。 BFCN 的发育受到围产期提供甲基的必需营养素胆碱的可用性的调节，怀孕期间胆碱摄入量的变化会导致胎儿大脑中 DNA 和组蛋白甲基化模式的显着改变。我们将通过测量胆碱摄入量对与整体 DNA 和组蛋白甲基化模式相关的 BFCN 基因表达模式的影响，对通过 FACS 纯化的小鼠 BFCN 在整个发育过程（胎儿到成年）进行首次此类研究。