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

项目摘要 选择性mRNA剪接（AS）是一个基本的过程，调节超过90%的基因表达。 人类蛋白质编码基因。AS在神经系统中的功能特别普遍， 与影响学习的多种神经系统疾病有关尽管AS与活动有关- 神经可塑性依赖的基因表达，没有系统的分析AS已经 在体内学习模型上进行。哺乳动物大脑的复杂性对此提出了挑战 类型的研究。因此，我们仍然不明白（1）学习在多大程度上使AS参与神经系统， 系统，（2）AS如何有助于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和特定基因的剪接异构体以产生学习行为。在这里，我们建议 在C中解决这些基本问题。优雅基本原理是， 氏梭线虫的神经系统被很好地表征，动态基因表达可以被描绘为 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剖析 与遗传和成像工具，以及发展和功能的基本属性， 神经系统在C.以及更复杂的动物。此外，许多形式 C.表现出的学习能力。秀丽线虫有着相似的行为特征和分子基础 那些由高等生物展示的。本项目的总体目标是描述AS如何调节 学习，并提供在许多疾病条件下大脑功能的神经缺陷的见解。 本项目的假设是AS通过调节神经元基因表达来调节神经功能 and produce生产learning学习.具体来说，我们将首先描述AS网络和拼接的全球模式 受我们实验室良好表征的学习范式调控的同种型。我们计划 系统地分析学习如何改变C. elegans 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功能， 神经活动和行为中保守分子的剪接异构体。该补助金是探索性的，因为它 (1)提出了第一个系统的分析AS在体内模型的学习和（2）介绍了概念 和技术进步来解决AS和学习行为之间的因果关系。拟议的工作是 重要的是，因为它（1）测试了AS的一个高度合理的功能，一个基本的基因表达过程 保守的真核生物，在学习，和（2）特点的机制，其中保守的 分子调节神经元基因表达和功能以产生学习。与此同时，我们的资助 在大量的初步结果，支持概念和技术生产力。的 本研究的结果将为其他系统中关于AS学习的研究提供关键和及时的见解 并进一步了解与异常AS相关的神经系统疾病中的学习缺陷。

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.