Epigenetic programming of brain development by choline nutrition

胆碱营养对大脑发育的表观遗传编程

• 批准号: 8144918
• 负责人: JAN Krzysztof BLUSZTAJN
• 金额: $ 19.61万
• 依托单位: BOSTON UNIVERSITY MEDICAL CAMPUS
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-09-20 至 2013-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8144918
• 关键词: Acetylcholine; Acetylcholinesterase; Adolescent; Adult; Age; Antibodies; Area; Attention; Basal Nucleus of Meynert; Brain; Capital; Cerebral cortex; Choline; Choline O-Acetyltransferase; Chromatin; DNA; DNA Methylation; DNA Methyltransferase; DNA Modification Methylases; 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

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）：累积证据表明表观遗传机制对于正常的脑发育至关重要，包括调节神经元和神经胶质，神经元细胞类型的规范和分化，电路和突触形成以及髓鞘形成。这些事件 - 在跨越妊娠和早期产后时间的关键时期发生 - 对必需营养素的供应非常敏感，尤其是对包括胆碱在内的代谢甲基供体的供应。该提案中描述的研究要检验的中心假设是，在这个关键时期，胆碱可用性的改变调节了通过涉及DNA和组蛋白甲基化变化的表观遗传机制的基础前脑胆碱能神经元（BFCN）的分化和表型。 BFCN构成了神经元电路的关键组成部分，这对于学习，记忆，注意力和睡眠过程所必需。我们已经开发了一种通过荧光激活的细胞分选的方法，使用转基因小鼠在胆碱乙酰转移酶基因的调节元件的控制下，通过荧光激活的细胞分类来表达增强的绿色荧光蛋白（EGFP）。在胚胎第16天到产后第30天之间，在各种发育阶段的纯化BFCN我们将确定：1）使用微阵列的全局转录组曲线； 2）用甲基化DNA免疫沉淀（MEDIP）的DNA甲基化模式，然后进行微阵列分析和3）染色质免疫沉淀（CHIP），抗体抗体对组蛋白3的特定赖氨酸残基的不同甲基化表位3（例如H3K9ME2; H3K4ME1; H3K4ME1; H3K4ME3）（例如，h3k4me1; h3k4me3）cip（例如）。这些数据将构成表观基因组的第一个特征及其与发展中纯神经元种群的转录组的相关性。我们将对小鼠的胎儿和后代进行类似的研究，其母亲在整个怀孕期间食用含有胆碱缺乏胆碱，缺乏胆碱或胆碱补充饮食的饮食，直到断奶为止。调查结果将提供有关BFCN发展里程碑以及通过妊娠和产后营养后营养胆碱摄入量与这些过程的调节相关的表观遗传过程的信息。 公共卫生相关性：基础前脑胆碱能神经元（BFCN）构成了神经元电路的关键组成部分，这对于学习，记忆，注意力和睡眠的过程是必需的。 BFCN发育受到偏甲基基本养分，胆碱的围产期可用性的调节，以及妊娠中胆碱摄入量的变化会导致胎儿脑中DNA和组蛋白甲基化模式的急剧修饰。我们将通过测量胆碱摄入量对BFCN基因表达与全球DNA和组蛋白甲基化模式相关的BFCN基因表达模式的影响，对FACS纯化的鼠BFCN进行第一项此类研究。