Regulation of Pet1/FEV binding and chromatin accessibility during serotonergic neuron development

血清素能神经元发育过程中 Pet1/FEV 结合和染色质可及性的调节

• 批准号: 10542353
• 负责人: Xinrui Zhang
• 金额: $ 5.27万
• 依托单位: CASE WESTERN RESERVE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-01-01 至 2023-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10542353
• 关键词: ATAC-seq; Adult; Anxiety; Architecture; Attention deficit hyperactivity disorder; Automobile Driving; Binding; Biological Assay; Brain; Cells; Central Nervous System; ChIP-seq; Characteristics; Chromatin; Chromatin Structure; Communication; Coupled; DNA; DNA Binding; DNA Methylation; Data Analyses; Dependence; Development; Developmental Gene; Disease; ETS Family Protein; Embryo; Embryonic Development; Emotional; Enhancers; Exhibits; Experimental Designs; Family; Functional disorder; Gene Expression; Gene Expression Profile; Genes; Genetic Transcription; Genomics; Growth; Hematopoietic stem cells; Heterochromatin; High-Throughput Nucleotide Sequencing; Human; Knockout Mice; Knowledge; Life; Link; Maintenance; Maps; Mediating; Mental Depression; Mental disorders; Molecular; Neurodevelopmental Disorder; Neurons; Neurosciences; Neurotransmitters; Oral; Organism; Orthologous Gene; Phylogenetic Analysis; Physicians; Physiological; Play; Population; Regulation; Regulator Genes; Regulatory Element; Role; SWI1; Schizophrenia; Scientist; Serotonin; Shapes; Sudden infant death syndrome; Synapses; Testing; Time; Training; Transposase; Up-Regulation; Writing; autism spectrum disorder; brain circuitry; cell type; chromatin immunoprecipitation; chromatin remodeling; embryonic stem cell; epigenetic profiling; epigenetic regulation; experience; experimental study; fetal; gene synthesis; genetic regulatory protein; histone modification; in vivo; insight; mouse genetics; neuron development; neuropsychiatric disorder; neuropsychiatry; neurotransmission; novel therapeutics; postmitotic; postnatal; postnatal period; promoter; training opportunity; transcription factor; transcriptome sequencing; transcriptomic profiling

PROJECT SUMMARY/ABSTRACT The neurotransmitter serotonin (5-HT) is implicated in the pathophysiology of many psychiatric and neurodevelopmental disorders, including anxiety, depression, autism, schizophrenia, attention- deficit/hyperactivity disorder, and compulsive disorders. How serotonin neurons mature and acquire their transmitter identity and adult characteristics remain poorly understood. During the embryonic and early postnatal period, lineage specific gene expression patterns of serotonin neurons are orchestrated by a network of developmentally critical transcription factors, including the ETS family transcription factor Pet1 (human ortholog FEV) that is essential for the establishment of serotonin neurotransmission. Intriguingly, we recently found Pet1 switches targets during fetal to early postnatal transition, from controlling the upregulation of 5-HT synthesis genes during fetal life to activating gene required for synaptic excitability during the postnatal period. This study investigates the hypothesis that the changes in the chromatin accessibility of cis-regulatory elements dictate the repertoire of transcriptional targets that are available for Pet1 regulation during development. Furthermore, I hypothesize that Pet1 binding also shapes chromatin architecture to direct the gene expression trajectories of developing serotonin neurons. To test these hypotheses, in Aim 1, I will map the global open chromatin landscape of serotonin neurons using Assay for Transposase Accessible Chromatin with high throughput sequencing (ATAC-seq) at multiple embryonic and early postnatal developmental time points, and investigate the relation of open chromatin to 5-HT neuron gene expression. In Aim 2, I will analyze the changes in Pet1 DNA occupancy during the same stages of serotonin neuron development as in Aim 1 using Pet1 chromatin immunoprecipitation coupled to high throughput sequencing (ChIP-seq), and determine the importance of Pet1 for developmentally critical chromatin remodeling or maintenance by performing ATAC- seq in Pet1 knockout mice. By elucidating how Pet1 dynamically regulates gene expression critical for the maturation of serotonin neurons, I will provide insight into the pathophysiology of the neuropsychiatric conditions in which serotonin gene expression is thought to be perturbed. Additionally, this study provides valuable training opportunity for me to gain technical proficiency in mouse genetics and transcriptomic and epigenetic profiling, broad conceptual knowledge in molecular neuroscience, and experience with experimental design, data interpretation, and oral and written communication that are crucial for my own growth as a physician-scientist in training.

项目概要/摘要 神经递质血清素 (5-HT) 与许多精神和疾病的病理生理学有关。 神经发育障碍，包括焦虑症、抑郁症、自闭症、精神分裂症、注意力障碍 缺陷/多动障碍和强迫症。血清素神经元如何成熟并获得它们 传播者身份和成人特征仍然知之甚少。在胚胎期和早期 出生后，血清素神经元的谱系特异性基因表达模式由网络精心策划 发育关键转录因子，包括 ETS 家族转录因子 Pet1（人类 直系同源 FEV）对于建立血清素神经传递至关重要。有趣的是，我们最近 发现 Pet1 在胎儿到产后早期的过渡期间转换目标，从控制 5-HT 的上调 胎儿时期合成基因，到出生后激活突触兴奋所需的基因。 本研究调查了以下假设：顺式调控染色质可及性的变化 元件决定了可用于 Pet1 调控的转录目标的全部。 发展。此外，我假设 Pet1 结合也塑造染色质结构以指导 发育中的血清素神经元的基因表达轨迹。为了测试这些假设，在目标 1 中，我将绘制 使用转座酶可及染色质检测分析血清素神经元的全局开放染色质景观 在多个胚胎和产后早期发育时间进行高通量测序 (ATAC-seq) 点，并研究开放染色质与 5-HT 神经元基因表达的关系。在目标2中，我将分析 与目标 1 相同的血清素神经元发育阶段中 Pet1 DNA 占据的变化 使用 Pet1 染色质免疫沉淀结合高通量测序 (ChIP-seq)，并确定 Pet1 对于发育关键染色质重塑或通过执行 ATAC 维护的重要性 Pet1 敲除小鼠中的 seq。通过阐明 Pet1 如何动态调节对 血清素神经元的成熟，我将深入了解神经精神的病理生理学 血清素基因表达被认为受到干扰的条件。此外，这项研究还提供了 宝贵的培训机会让我获得小鼠遗传学和转录组学方面的技术熟练程度 表观遗传分析、分子神经科学的广泛概念知识以及实验经验 设计、数据解释以及口头和书面交流，这些对于我作为一个人的成长至关重要 接受培训的医师科学家。