Systematic and functional analysis of alternative mRNA splicing in an in vivo model of learning

项目总结 选择性信使核糖核酸剪接(AS)是一个基本的过程，它调节超过90%的 人类蛋白质编码基因。AS在神经系统中的功能特别普遍，一直以来 牵涉到损害学习的多种神经疾病。尽管AS与活动有牵连- 神经可塑性背后的依赖基因表达，尚未对AS进行系统分析 在活体学习模型上执行。哺乳动物大脑的复杂性对此提出了挑战 研究类型。因此，我们仍然不明白(1)学习在多大程度上与神经有关 系统，(2)AS如何促进学习诱导的神经元基因表达的变化，以及(3)如何 学习调节AS并拼接特定基因的异构体，以产生习得行为。在这里，我们建议 来解决线虫的这些基本问题。其基本原理是连线和基因构成 秀丽线虫神经系统的特征，可以用来描述动态基因表达 无论是整个大脑还是单个神经元，基因在学习中的功能都可以在细胞分辨率上进行解剖。 与遗传和成像工具，以及基本属性的发展和功能的 线虫和更复杂的动物之间的神经系统保存得很好。此外，许多形式 线虫表现出的学习能力具有相似的行为特征和分子基础 那些由高等有机体表现出来的。这个项目的总体目标是描述AS如何监管 学习和提供许多疾病条件下大脑功能的神经缺陷的洞察力。 该项目的假设是AS调节神经元基因表达，从而调节神经功能。 并产生学习。具体地说，我们将首先描述AS网络和拼接的全局模式 由我们实验室的学习范例很好地描述的异构体。我们计划 系统分析学习如何改变线虫中表达的所有基因的剪接或异构体使用 神经系统。接下来，我们将使用遗传扰动来解决学习调节的因果函数 剪接神经活动和行为中保守分子的异构体。这笔赠款是探索性的，因为它 (1)首次在活体学习模型中对AS进行了系统分析；(2)介绍了概念 和技术进步，以解决自闭症和学习行为之间的因果联系。建议的工作是 意义重大，因为它(1)测试了AS的一个非常可信的功能，这是一个基本的基因表达过程 在学习中在真核生物中保守的，以及(2)具有保守性的机制的 分子调节神经元基因的表达和功能，以产生学习。与此同时，我们的拨款是建立在 基于大量支持概念和技术生产力的初步结果。这个 这项研究的结果将为在其他系统中学习AS的研究提供关键和及时的见解 并促进对与异常AS相关的神经系统疾病的学习缺陷的理解。

PROJECT SUMMARY Alternative mRNA splicing (AS) is a fundamental process that regulates the expression of more than 90% of human protein-coding genes. The function of AS in the nervous system is particularly prevalent and has been implicated in multiple neurological disorders that impair learning. Although AS is implicated in activity- dependent gene expression underlying neural plasticity, no systematic analysis on AS has been performed on an in vivo learning model. The complexity of the mammalian brain poses challenges to this type of studies. Thus, we still do not understand (1) to what extent learning engages AS in the nervous system, (2) how AS contributes to learning-induced changes in neuronal gene expression, and (3) how learning modulates AS and splice isoforms of specific genes to generate learned behavior. Here, we propose to address these fundamental questions in C. elegans. The rationale is that the wiring and genetic make-up of the C. elegans nervous system are well characterized, dynamic gene expression can be profiled for the whole brain or individual neurons, functions of genes in learning can be dissected at the cellular resolution with genetic and imaging tools, and the fundamental properties of the development and function of the nervous system are well conserved between C. elegans and more complex animals. In addition, many forms of learning exhibited by C. elegans share similar behavioral characteristics and molecular underpinnings with those displayed by higher organisms. The overall goal of this project is to characterize how AS regulates learning and to provide insights into neurological defects in brain function under many disease conditions. The hypothesis of this project is that AS regulates neuronal gene expression to modulate neural function and produce learning. Specifically, we will first characterize the global patterns of AS network and splice isoforms that are regulated by a learning paradigm well-characterized in our laboratory. We plan to systematically analyze how learning alters splicing or isoform usage of all genes expressed in the C. elegans nervous system. Next, we will use genetic perturbations to address the causal function of learning-regulated splice isoforms of conserved molecules in neural activity and behavior. The grant is exploratory, because it (1) presents the first systematic analysis of AS in an in vivo model of learning and (2) introduces conceptual and technical advances to address causal links between AS and learning behavior. The proposed work is significant, because it (1) tests a highly plausible function of AS, a fundamental gene expression process conserved in eukaryotes, in learning, and (2) characterizes the mechanisms whereby AS of conserved molecules regulates neuronal gene expression and function to produce learning. Meanwhile, our grant is built on a substantial amount of preliminary results that support conceptual and technical productivity. The outcome of this study will provide critical and timely insights into the studies on AS in learning in other systems and advance understanding of learning defects in neurological diseases associated with aberrant AS.