Brain serotonin neuron gene regulatory networks and chromatin architecture

大脑血清素神经元基因调控网络和染色质结构

• 批准号: 10295748
• 负责人: EVAN S DENERIS
• 金额: $ 44.01万
• 依托单位: CASE WESTERN RESERVE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-02-01 至 2024-10-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10295748
• 关键词: ADRA1B gene; ATAC-seq; Address; Anxiety; Architecture; Binding Sites; Biological Assay; Brain; Cell Nucleus; Chromatin; Defect; Dependence; Development; Disease; Environmental Risk Factor; G-Protein-Coupled Receptors; GCHFR gene; Gene Expression; Gene Expression Profile; Gene Expression Regulation; Genes; Genetic; Genetic Transcription; Investigation; Knock-out; Knowledge; Label; Life; Maps; Mental Depression; Molecular; Mus; Neurodevelopmental Disorder; Neurons; Neurotransmitters; Pattern; Plant Roots; Property; Protocols documentation; Regulation; Regulator Genes; Research; Schizophrenia; Series; Serotonin; Stress; Synapses; Testing; Tn5 transposase; Up-Regulation; anxiety-related disorders; autism spectrum disorder; brain health; chromatin remodeling; critical period; environmental stressor; fetal; gene regulatory network; gene synthesis; interest; neural circuit; neuron development; neuropsychiatric disorder; novel; postnatal; postnatal period; preservation; spatiotemporal; transcription factor; transcriptome; transcriptome sequencing; young adult

Precise spatiotemporal control of gene expression is crucial throughout life for maturation of postmitotic neuronal function and preservation of brain health. Although continuously expressed neuronal transcription factors termed terminal selectors, such as Pet1 and Lxm1b, are key regulators of gene expression across fetal and postnatal stages of life the underlying molecular mechanisms through which they control stage specific neuronal gene expression are poorly understood. Here, we aim to fill this gap by comprehensively investigating terminal selector function in postmitotic serotonin (5-HT) neurons. Pet1 and Lmx1b’s control of serotonergic gene expression is of broad interest as 5-HT has wide-ranging modulatory effects on central neural circuitry and altered serotonergic gene expression has been implicated in several developmental neuropsychiatric disorders including depression, stress-related anxiety, autism, OCD, and schizophrenia. The studies proposed here are motivated, in part, by our discovery of changing dependencies of continuously expressed 5-HT neuron terminal effector genes on Pet1 as 5-HT neurons mature. We unexpectedly found that Pet1’s terminal selector control of 5-HT synthesis genes is largely switched off in the early postnatal period and instead Pet1 switches to controlling the upregulation of neurotransmitter GPCR genes, Htr1a, Adra1b, that are needed for afferent synaptic modulation of 5-HT neuron excitability. These recent observations have led us to suggest a new principle that we have termed “terminal selector target switching”. We suggest that as postmitotic neurons progress through life, continuous terminal selector regulated transcription is not static, as is the prevailing assumption. Instead, we hypothesize that terminal selector regulated transcription is highly dynamic in which regulatory factor interactions with target genes are remodeled as postmitotic neurons mature. We hypothesize the remodeling of regulatory interactions is rooted in the temporal remodeling of postmitotic neuronal chromatin architecture. In Aim 1, we will investigate Lmx1b’s control of early postnatal 5-HT gene expression to determine whether postmitotic target switching is a general property of 5-HT terminal selectors. In Aim 2, we will use our newly developed 5HT-ATAC-seq protocol to uncover the temporal dynamics of 5-HT open chromatin as 5-HT neurons develop and mature from fetal to early postnatal stages of life. Specifically, we will address these questions: Do open chromatin states undergo maturational changes in parallel with the maturation of 5-HT neuron transcriptomes? Do switches in target dependence result from gene specific changes in open chromatin? Aim 3 will investigate a plausible potential mechanism through which developing 5-HT gene expression patterns are controlled: Terminal selectors, Pet1 and Lmx1b, directly act to control the maturation of 5-HT neuronal open chromatin states. Understanding how terminal selectors control postmitotic neuronal gene expression may illuminate potential neurodevelopmental disease mechanisms that cause aberrant neuronal gene expression and defects in neuronal maturation.

基因表达的精确时空控制对于蒙脱病的成熟至关重要 神经元功能和脑健康的保存。尽管连续表达神经元转录 所谓的末端选择器（例如PET1和LXM1B）是胎儿基因表达的关键调节剂 生活的产后阶段，它们控制特定阶段的基本分子机制 神经元基因表达知之甚少。在这里，我们旨在通过全面调查来填补这一空白 终末选择器功能在神经后血清素（5-HT）神经元中。 PET1和LMX1B的血清素能控制 基因表达具有广泛的兴趣，因为5-HT对中央神经回路具有广泛的调节作用 以及多种发育神经精神病学中实施了血清素能基因表达的改变 包括抑郁症，与压力相关的动画，自闭症，强迫症和精神分裂症在内的疾病。研究提出 这是我们发现不断变化的连续表达5-HT的依赖性的部分动机 神经元末端效应子基因在PET1上为5-HT神经元成熟。我们意外地发现Pet1的终端 5-HT合成基因的选择器控制在产后早期主要关闭，而是PET1 切换到控制神经递质GPCR基因HTR1A，ADRA1B的上调，这是需要的 令人兴奋的5-HT神经元的传入突触调节。这些最近的观察结果使我们提出了一个 我们称为“终端选择器目标切换”的新原则。我们建议作为有丝分裂后神经元 通过生活的进展，连续终端选择器调节的转录不是静态的， 假设。相反，我们假设终端选择器调节的转录是高度动态的 调节因子与靶基因的相互作用被重塑为有丝分裂后神经元成熟。我们假设 调节相互作用的重塑植根于有丝分裂后染色质的临时重塑 建筑学。在AIM 1中，我们将研究LMX1B对早期产后5-HT基因表达的控制 确定有丝分裂后目标转换是否是5-HT终端选择器的一般特性。在AIM 2中，我们 将使用我们新开发的5HT-ATAC-SEQ协议来揭示5-HT Open的临时动态 染色质作为5-HT神经元的发育和从胎儿到早期生命的早期阶段成熟。具体来说，我们会的 解决以下问题：开放染色质状态与与之并联的成熟变化 5-HT神经元转录组的成熟？ DO转换目标依赖性是基因特异性的 开放染色质的变化？ AIM 3将研究一种合理的潜在机制 5-HT基因表达模式受到控制：端子选择器PET1和LMX1B直接作用以控制 5-HT神经元开放染色质状态的成熟。了解终端选择器如何控制蒙脱病后 神经元基因表达可能会阐明引起潜在的神经发育疾病机制 神经元成熟的异常神经元基因表达和缺陷。