Transcriptional regulation of post-embryonic neuronal maturation

项目摘要/摘要 在神经系统发育和成熟的复杂和错综复杂的时间序列中， 通过遗传和环境对动态监管过程的重大和持续的破坏 风险可能导致神经发育障碍。神经科学研究对我们的 胚胎和围产期包括神经元在内的早期神经发育的认识 规范、神经元迁移、轴突引导/生长和突触发生。从早期开始的时期 从出生后到成年，对于成熟的大脑的正常发育同样至关重要。 在这些时期观察到结构和行为的可塑性。然而，人们对此知之甚少 出生后发育过程中神经元成熟的分子调控机制。在第一个 在我博士后工作的一部分，我开发并优化了一种分离单个神经元类型的方法，用于基因组学 侧写。使用这种方法，结合线虫系统的遗传适宜性，我将 单个神经元类型在出生后早期的转录组和染色质可及性 发展到青年期(目标1)。在目标2中，使用来自目标1的性能分析数据作为入口点，i 将采取双管齐下的方法来确定神经元成熟的新调节因子。首先，我将共同开发和 利用生物信息学工具从发育调控基因中提取常见的顺式调控基序 并预测候选转录调控因子。第二，使用有效的发育调节基因 记者在Aim 1，我将进行无偏见的正向基因筛查，以确定神经元的新调节因子 成熟。在目标3中，我将使用先进的神经技术工具来研究环境刺激的作用 (感觉诱导的神经元活动)对神经元成熟的影响。首先，我将研究急性和慢性抑制是如何 靶神经元在不同发育窗口期的活性影响单个神经元的发育 转录本和染色质的可及性。此外，调节神经元的通路之间的相互作用 通过环境诱导的活动与内在的遗传机制的成熟将被检验。 使用目标2中的方法。拟议的研究将增加我们对遗传和 支持正常出生后神经发育的环境调节机制，改善我们的 了解神经发育障碍的病因，并揭示新的治疗靶点。 该奖项的培训阶段在奥利弗·霍伯特博士的实验室进行，在联合- 哥伦比亚大学Harmen BusSemaker博士和Stavros Lomvardas博士的指导，概述了 获得技术和专业技能的全面计划，使我能够过渡到 独立研究岗位。该项目的顺利完成将为今后的工作提供平台 旨在了解基因-环境在神经转录调控中相互作用的实验 系统的开发和功能。 。

PROJECT SUMMARY/ABSTRACT 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 the first part of my post-doctoral work, 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 will profile the transcriptome and chromatin accessibility of single neuron types throughout early post-natal development to young adulthood (Aim 1). In Aim 2, using the profiling data from Aim 1 as an entry point, I will take a 2-pronged approach to identify novel regulators of neuronal maturation. First, I will co-develop and implement bioinformatics tools to extract common cis-regulatory motifs from developmentally regulated genes and predict candidate transcriptional regulators. Second, using validated developmentally-regulated gene reporters in Aim 1, I will conduct unbiased forward genetic screens to identify novel regulators of neuronal maturation. In Aim 3, 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 2. 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 training phase of the award, conducted in the laboratory of Dr. Oliver Hobert and under the co- mentorship of Dr. Harmen Bussemaker and Dr. Stavros Lomvardas at Columbia University, outlines a comprehensive plan for the acquisition of technical and professional skills that will enable my transition to an independent research position. 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. .