RNA methylation in synaptic plasticity and drug-seeking

RNA甲基化在突触可塑性和药物寻找中的作用

基本信息

批准号：
10159882
负责人：
Xiaoxi Zhuang
金额：
$ 52.66万
依托单位：
UNIVERSITY OF CHICAGO
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-09-01 至 2022-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10159882
关键词：
Addictive Behavior Affect Agonist Animal Model Binding Binding Proteins Biological Biological Assay Brain Cell Culture Techniques Cell physiology Cells Cellular biology Cocaine Coculture Techniques Corpus striatum structure Cues Cyclic AMP Cyclic AMP-Dependent Protein Kinases DLG4 gene DNA DNA Methylation Data Dendrites Dendritic Spines Desire for food Dopamine Dopamine D1 Receptor Drug Addiction Epigenetic Process Explosion Fluorescent in Situ Hybridization Food Gene Expression Genetic Transcription Impairment Knock-out Label Learning Long-Term Potentiation Mass Spectrum Analysis Mental Depression Messenger RNA Methods Methylation Methyltransferase Modeling Modification Mus Mutant Strains Mice Neurons Nucleus Accumbens Pathway interactions Pharmaceutical Preparations Play Protein Biosynthesis Proteins Psychological reinforcement Publishing RNA RNA Biochemistry RNA Splicing RNA methylation Reader Research Rest Rewards Role Self Administration Signal Transduction Slice Spatial Distribution Specificity Synapses Synaptic plasticity Testing Translating Translations base cocaine exposure cocaine self-administration conditional knockout density drug seeking behavior epitranscriptome experimental study glutamatergic signaling histone modification in vivo mouse genetics neuronal cell body overexpression protein transport response single molecule transcriptome sequencing

项目摘要

Abstract Aberrant synaptic plasticity is a key underlying biological mechanism of drug addiction. The long-lasting synaptic changes are dependent on changes in gene expression and evidence support the involvement of epigenetic mechanisms. However, histone modifications, DNA methylation or DNA hydroxymethylation affects gene expression at transcriptional level, which is not only slow in affecting protein synthesis but also lacks synaptic specificity. RNA N6-methyladenosine (m6A) modification has been found to significantly affect RNA splicing, export, localization, translation efficiency and stability. All there are key factors in regulating translation and potentially localized translation in synapses. Both our preliminary data and published studies suggest that deficiency in m6A dependent pathways significantly impairs neuronal function including dopamine signaling and dopamine dependent learning. Therefore we propose to focus on the role of m6A modification of mRNAs in synaptic plasticity and in drug addiction models. We will use mice with the m6A methyltransferase METTL14 deleted in D1 positive striatal neurons in the nucleus accumbens. In Aim 1, we will test the hypothesis that mutant mice have impaired appetitive Pavlovian learning and impaired cue-induced reinstatement of cocaine self-administration. In Aim 2, we will record from brain slices and test if mutant mice have impaired corticostriatal plasticity and diminished changes in corticostriatal synaptic strengths caused by cocaine exposure. In Aim 3, we will determine how m6A RNA methylation regulates synaptic protein translation in responses to drug challenges and other changes on neuronal activity. We will first examine if cocaine exposure in vivo affects level of m6A RNA methylation and identify downstream targets. The rest of the proposed experiments will examine the degree to which m6A RNA methylation affects protein translation in the soma vs. dendrites where local translation is under control of synapses. We will use dissociated neuronal cortical-striatal co-cultures from METTL14 knockout and control mice.

抽象的异常的突触可塑性是药物成瘾的一个关键的潜在生物学机制。持久的突触变化取决于基因表达的变化，有证据支持表观遗传机制。然而，组蛋白修饰、DNA 甲基化或 DNA 羟甲基化会影响基因在转录水平表达，不仅影响蛋白质合成缓慢，而且缺乏突触特异性。 RNA N6-甲基腺苷 (m6A) 修饰被发现显着影响 RNA 拼接、导出、本地化、翻译效率稳定。规范翻译的关键因素以及突触中潜在的本地化翻译。我们的初步数据和已发表的研究都表明 m6A 依赖性通路的缺陷会显着损害神经元功能，包括多巴胺信号传导和多巴胺依赖性学习。因此，我们建议重点关注 mRNA 的 m6A 修饰在突触可塑性和药物成瘾模型。我们将使用具有 m6A 甲基转移酶 METTL14 的小鼠伏隔核中的 D1 阳性纹状体神经元被删除。在目标 1 中，我们将检验以下假设：突变小鼠巴甫洛夫食欲学习能力受损，提示诱导的可卡因恢复能力受损自我管理。在目标 2 中，我们将记录大脑切片并测试突变小鼠是否受损可卡因引起的皮质纹状体可塑性和皮质纹状体突触强度变化减弱接触。在目标 3 中，我们将确定 m6A RNA 甲基化如何调节突触蛋白翻译对药物挑战和神经元活动的其他变化的反应。我们将首先检查是否接触可卡因体内影响 m6A RNA 甲基化水平并识别下游靶标。其余建议实验将检查 m6A RNA 甲基化对体细胞和体外蛋白质翻译的影响程度。局部翻译受突触控制的树突。我们将使用分离的神经元皮质纹状体 METTL14 敲除小鼠和对照小鼠的共培养物。