Functional Role for H3 Serotonylation During Critical Periods of Postnatal Brain Development and Plasticity

项目摘要/摘要 5-羟色胺能(5-羟色胺)系统与多种神经发育和神经精神疾病有关。 现象，包括情绪和应激反应的调节。虽然5HT的行动被认为是 完全通过5-羟色胺受体及其突触效应介导，最近的研究表明 中缝背侧5-羟色胺能神经元和前脑靶神经元存在5-羟色胺核池。我们的 实验室已经证实，5-羟色胺与组蛋白H3-形成共价键，从而产生H3-谷氨酰胺5-羟色胺 (H3Q5ser)--一个被称为H3血清素基化的过程。我们进一步证明了H3的5-羟色胺基化起着关键作用 在建立正常的胚胎和成年大脑转录可塑性模式中的调节作用。然而， H3 5羟色胺在出生后早期脑发育中的功能作用，包括 神经可塑性的关键时期，在很大程度上还没有被探索，以及环境刺激的影响 (异常或其他)在早期生命中对这种修饰的影响仍然未知。我假设H3 在整个出生后的大脑中，5-羟色胺的变化轨迹--以一种区域特有的方式--来控制关键的 神经发育基因表达和早期生活压力的各个方面打乱了这些模式，直接影响 易受压力相关行为异常的影响。在伊恩·梅兹和埃里克·内斯特勒博士的指导下 在西奈山的伊坎医学院，我将以三个不同的目标来阐述这一假设，使用 方法多种多样。在目标1中，我将从功能上评估出生后H3 5羟色胺的动态变化 发展，研究早期生活压力如何扰乱这些模式，并阐明其中的因果关系 表基因组与组蛋白策略整合对基因表达调控的新型组蛋白修饰 转录切分方法(即RNA测序和切割和运行)。在目标2中，我将描述中断是如何 出生后5-羟色胺相关基因的表达直接导致脑回路的改变，从而导致 使用先进的基因编辑技术和 行为分析。在目标3中，我将描述出生后大脑中H3 5-羟色胺的全脑模式 使用全脑清理和成像技术(即，)来开发和响应早期的类似压力。 IDISCO+)。总体而言，这个项目将为h3 5羟色胺控制大脑的方式提供新的见解。 这一翻译后标记的发展和破坏导致异常的机制 病理生理状态。

Project Summary/Abstract The serotonergic (5HTergic) system is implicated in a wide range of neurodevelopmental and neuropsychiatric phenomena, including the regulation of mood and stress reactivity. While 5HT actions have been assumed to be mediated exclusively through 5HT receptors and their synaptic effects, recent studies have demonstrated the presence of nuclear pools of 5HT in dorsal raphe serotonergic neurons and forebrain target neurons. Our laboratory has established that 5HT forms covalent bonds with histone H3—resulting in H3 glutamine 5 serotonin (H3Q5ser)–a process known as H3 serotonylation. We have further shown that H3 serotonylation plays key regulatory roles in establishing normal embryonic and adult patterns of brain transcriptional plasticity. However, functional roles for H3 serotonylation during early post-natal brain development, time points encompassing critical periods of neural plasticity, have largely been unexplored, and the impact of environmental stimuli (aberrant or otherwise) on this modification during early life remains unknown. I hypothesize that H3 serotonylation trajectories vary – in a region-specific manner – across the post-natal brain to control critical aspects of neurodevelopmental gene expression and early life stress disrupts these patterns directly influencing vulnerability to stress-related behavioral abnormalities. Under the mentorship of Drs. Ian Maze and Eric Nestler at the Icahn School of Medicine at Mount Sinai, I will address this hypothesis with three distinct aims using a variety of approaches. In Aim 1, I will functionally assess H3 serotonylation dynamics during post-natal development, examine how early life stress disrupts these patterns, and elucidate the causal relationship of this novel histone modification on gene expression regulation using strategic integration of epigenomic and transcriptomic approaches (ie. RNA sequencing and CUT&RUN). In Aim 2, I will characterize how disruptions in post-natal serotonylation-associated gene expression directly result in alterations in brain circuitry leading to increased vulnerability to stress-related behavioral abnormalities using advanced gene-editing techniques and behavioral analyses. In Aim 3, I will characterize brain-wide patterns of H3 serotonylation across post-natal brain development and in response to early like stress using whole brain clearing and imaging techniques (ie. iDISCO+). Overall, this project will provide novel insight into the ways that h3 serotonylation controls brain development and the mechanisms by which disruptions to this posttranslational mark cause aberrant pathophysiological states.