Transcriptional regulation of post-embryonic neuronal maturation

项目总结 在神经系统发育和成熟的复杂和错综复杂的时间序列中，意义重大 遗传和环境风险对动态监管过程的持续干扰可能 会导致神经发育障碍。神经科学研究对我们理解 胚胎和围产期的早期神经发育，包括神经元规格、神经元 迁移、轴突引导/生长和突触发生。从出生后早期到年轻的时期 成年期对于成熟大脑的正常发育和广泛的结构和行为同样重要。 在这些时期观察到了可塑性。然而，对分子调控的了解要少得多。 出生后发育过程中神经元成熟的机制。在我的博士后工作中/K99 在获奖阶段，我开发和优化了一种分离单个神经元类型的方法，用于基因组学图谱分析。 使用这种方法，结合线虫系统的遗传适宜性，我描绘了 转录组和染色质在神经系统在出生后早期发育过程中的可及性 并发现了一条由microRNA lin-4(脊椎动物中的mir-125)启动的保守途径， 它控制了许多但不是所有发育调节的、神经元特异性基因表达的变化， 这是神经元成熟的基础。在目标1中，使用基因组图谱数据作为切入点，我将采取2- 确定神经元成熟的额外的、新的调节因子的分叉方法。首先，我将实施 生物信息学工具从发育调节基因中提取常见顺式调控基序 预测/测试候选转录调控因子。第二，使用有效的发育调节基因 记者从我的简介中，我将进行无偏见的前向基因筛查，以确定新的调节者 神经元成熟。在目标2中，我将使用先进的神经技术工具来研究环境的作用 刺激(感觉诱导的神经元活动)对神经元成熟的影响。首先，我将检查急性和慢性 在不同的发育窗口期抑制靶神经元的活动会影响单个神经元 发育转录本和染色质可及性。此外，通路之间的相互作用 通过环境诱导的活动与内在遗传机制调控神经元成熟 将使用目标1中的方法进行研究。拟议的研究将增加我们对 遗传和环境调节机制支持正常的产后神经发育，改善 我们对神经发育障碍的病因的理解，并揭示了新的治疗靶点。这个 该项目的成功完成将为今后的实验提供一个平台 基因-环境在神经系统发育和功能的转录调控中相互作用。

PROJECT SUMMARY During the complex and intricately-timed sequence of nervous system development and maturation, significant and persistent disruptions to the dynamic regulatory process through genetic and environmental risks can result in neurodevelopmental disorders. Neuroscience research has contributed much to our understanding of early neural development in the embryonic and perinatal periods including neuronal specification, neuronal migration, axonal guidance/outgrowth, and synaptogenesis. The period from early post-natal life to young adulthood is equally critical for proper development of the mature brain, as extensive structural and behavioral plasticity are observed during these periods. However, much less is known about the molecular regulatory mechanisms underlying neuronal maturation during post-natal development. In my post-doctoral work/K99 phase of the award, I developed and optimized a method to isolate single neuron types for genomics profiling. Using this method, in combination with the genetic amenability of the C. elegans system, I profiled the transcriptome and chromatin accessibility of the nervous system throughout early post-natal development to young adulthood and identified a conserved pathway, initiated by the microRNA lin-4 (mir-125 in vertebrates), that controlled many but not all developmental-regulated, neuron-specific gene expression changes that underlie neuronal maturation. In Aim 1, using the genomic profiling data as an entry point, I will take a 2- pronged approach to identify additional, novel regulators of neuronal maturation. First, I will implement bioinformatics tools to extract common cis-regulatory motifs from developmentally regulated genes and predict/test candidate transcriptional regulators. Second, using validated developmentally-regulated gene reporters from my profiling, I will conduct unbiased forward genetic screens to identify novel regulators of neuronal maturation. In Aim 2, I will use advanced neurotechnology tools to examine the role of environmental stimuli (sensory-induced neuron activity) on neuronal maturation. First, I will examine how acute and chronic inhibition of activity in target neurons, during different developmental windows, affects single neuron developmental transcriptomes and chromatin accessibility. In addition, the interaction between pathways regulating neuronal maturation through environmentally-induced activity versus intrinsic genetic mechanisms will be examined using approaches as in Aim 1. The proposed research will increase our understanding of the genetic and environmental regulatory mechanisms underlying normal post-natal neurodevelopment, improve our understanding of the etiology of neurodevelopmental disorders, and reveal novel therapeutic targets. The successful completion of this project will provide a platform for future experiments aimed at understanding gene-environment interplay in the transcriptional regulation of nervous system development and function.