miR-137 Regulation of Intrinsic Excitability

项目摘要 MiRNAs是一种小的、内源性的、非编码的RNA，越来越多地被证明发挥着重要的作用 在基因表达调控中的作用。MiR-137是一种高度保守的富含大脑的miRNA，已被连接到 多发性大脑疾病，包括精神分裂症、自闭症谱系障碍(ASD)、双相情感障碍和 智力残疾(ID)。我们的长期目标是确定miR-137影响内源性的所有基因靶点 神经元兴奋性，并了解这些调节事件如何、何时和在哪里对信号起作用 神经系统的可塑性。在拟议的研究中，我们将首先检查内在和突触如何 神经元的活性随着miR-137表达水平的改变而改变，然后识别 在体内导致兴奋性改变的MIR-137。利用果蝇作为实验系统，我们将 结合强大的分子遗传学工具，包括CRISPR-Cas9技术，制造定点基因组 变化，用生物化学和电生理学来检查：1)神经元的内在电活动是如何 2)离子通道基因，包括Kv4/Shal、Kv3/Shaw、Kcnh/EAG、Cav3/Ca-α1T、 和NAV1/对离子通道基因，以及其他与活动相关的靶点，由miR-137调节以影响 活体内神经元兴奋性。

Project Summary miRNAs are small, endogenous, non-coding RNAs that are increasingly being shown to play important roles in regulating gene expression. miR-137 is a highly conserved brain-enriched miRNA that has been linked to multiple brain disorders, including schizophrenia, autism spectrum disorder (ASD), bipolar disorder, and intellectual disability (ID). Our long-term goal is to identify all of the gene targets of miR-137 that affect intrinsic neuronal excitability, and understand how, when, and where these regulatory events contribute to signaling and plasticity in the nervous system. In the proposed studies, we will first examine how intrinsic and synaptic activity in neurons is changed in response to altered expression levels of miR-137, then identify the targets of miR-137 that contribute to altered excitability in vivo. Using Drosophila as an experimental system, we will combine powerful molecular-genetic tools, including CRISPR-Cas9 technology to make site-directed genomic changes, with biochemistry and electrophysiology to examine: 1) how intrinsic electrical activity in neurons is regulated by miR-137, and 2) if ion channel genes, including Kv4/Shal, Kv3/Shaw, KCNH/eag, Cav3/Ca-α1T, and Nav1/para ion channel genes, as well as other activity-related targets, are regulated by miR-137 to affect neuronal excitability in vivo.