The role of a novel viral-like signalling pathway in synaptic plasticity and neurological disorders

新型病毒样信号通路在突触可塑性和神经系统疾病中的作用

• 批准号: 9802983
• 负责人: Travis Thomson
• 金额: $ 36.64万
• 依托单位: UNIV OF MASSACHUSETTS MED SCH WORCESTER
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-08-15 至 2024-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9802983
• 关键词: 3' Untranslated Regions; Actins; Address; Amino Acid Sequence; Antibodies; Behavior; Binding; Biological Models; Capsid; Cells; Co-Immunoprecipitations; Communication; Cytoskeleton; Degenerative Disorder; Development; Down-Regulation; Drosophila genus; Encapsulated; Epilepsy; Exhibits; Genes; Genetic; Genome; Goals; Grant; Homologous Gene; Human; Infectious Agent; Learning; Mammals; Mediating; Memory; Messenger RNA; Molecular; Multivesicular Body; Muscle; Nervous system structure; Neuromuscular Diseases; Neuromuscular Junction; Neuronal Plasticity; Neurons; Pathway interactions; Physiological; Play; Presynaptic Terminals; Proteins; RNA; RNA Interference; RNA immunoprecipitation sequencing; Reagent; Reporting; Research; Reticulum; Retrotransposon; Role; Schizophrenia; Selfish Genes; Signal Pathway; Signal Transduction; Spinocerebellar Ataxias; Synapses; Synaptic Vesicles; Synaptic plasticity; System; Testing; Transcript; Transfer RNA; Travel; Uncertainty; Ursidae Family; Viral; Viral Genome; Virus; WNT Signaling Pathway; Work; experimental study; extracellular vesicles; fly; gag Gene Products; human disease; intercellular communication; knock-down; nervous system disorder; neuromuscular; neuron development; neurotransmission; novel; particle; postsynaptic; presynaptic; protein function; trafficking; viral RNA

We have discovered a novel viral-like signaling pathway associated with extracellular vesicles (EV). We found the Drosophila homolog of ARC (Actin-Regulated Cytoskeleton-associated protein) (darc1), is present in EVs as both an mRNA and protein. ARC is a master regulator of synaptic plasticity in the nervous system of mammals and is crucial for learning and memory. dArc1 bears a domain resembling retroviral/retrotransposon Gag-like proteins that multimerizes into a capsid that packages viral RNA. Our work shows dARC1 forms a capsid, associates with its own RNA, and then transports the darc1 transcript across the synapse. The transfer of dArc1 is needed for activity-dependent plasticity at the fly neuromuscular junction (NMJ). Besides dArc1, it is unknown whether other genes are in this viral-like pathway. We address this uncertainty in Aim 1. Here we describe our plan to identify other Gag-like proteins in EVs, and we have already found another Gag protein enriched in EVs, that is encoded by the retrotransposon Copia. We have found that Copia transfers across the synaptic bouton. When copia is knocked down at the NMJ this strikingly leads to increased plasticity. This is the opposite of darc1, where we reported a decrease in plasticity. In Aim 2 we focus on what cargoes are co-transferring with dArc1 and Copia. We have identified through co-immunoprecipitation, mRNAs and proteins that associate with dArc1 and Copia. As to how the transfer of Arc occurs, we have found that the dArc1 3’untranslated region (UTR) is necessary and sufficient for the transfer of dArc1 across synaptic boutons. We are now testing if the dArc1 3’UTR directs the loading of dArc1 into EVs. As well, we propose experiments to understand how darc1 and copia mediate synaptic plasticity. We have co-immunoprecipitated dArc1 and Copia to identify potential interactors, and we will take a candidate approach to find genetic interactors. In preliminary work we found that dArc1 is needed for proper WNT pathway signaling at the NMJ. Additionally, we observe that Copia and dArc1 bind to some of the same proteins and mRNAs, suggesting that they may be antagonistic to each other, thus potentially explaining their seemingly opposite roles in mediating plasticity. Through this grant we will expand our understanding of EV trafficking and synaptic plasticity, while describing a novel physiological function of a retrotransposon in neuronal communication.

我们发现了一种与细胞外蔬菜相关的新型病毒状信号通路（EV）。我们 发现弧菌（肌动蛋白调节的细胞骨架相关蛋白）（DARC1）的果蝇同源物为 在电动汽车中作为mRNA和蛋白质存在。 ARC是神经系统突触可塑性的主调节器 哺乳动物系统，对于学习和记忆至关重要。 darc1带有一个类似于 逆转录病毒/逆转录座子类GAG样蛋白多含量地将其包装为病毒RNA包装的衣壳。我们的 工作表明Darc1形成了一个Capsid，与自己的RNA相关联，然后运输DARC1成绩单 穿过突触。 DARC1的转移是需要活动依赖性可塑性的转移 神经肌肉连接（NMJ）。除了darc1外，还不知道其他基因是否在这种病毒状态下 路径。我们在目标1中解决了这种不确定性。在这里，我们描述了我们的计划，以识别其他类似插科打的 电动汽车中的蛋白质，我们已经发现了另一种富含电动汽车的GAG蛋白，该蛋白质由 Retrotransposon copia。我们发现，Copia跨过突触式花道。何时Copia 击倒NMJ，这显着导致可塑性增加。这与darc1相反，那里 我们报告了可塑性的降低。在AIM 2中，我们专注于与Darc1共同转移的货物和 帕西亚。我们已经通过与DARC1相关的共免疫沉淀，mRNA和蛋白质确定 和Copia。至于弧的转移是如何发生的，我们发现Darc1 3'untranslated区域（UTR） 对于将DARC1转移到突触胸子上是必要且足够的。我们现在正在测试是否 Darc1 3’utr将Darc1的负载引导到电动汽车中。同样，我们提出了实验，以了解如何 DARC1和COPIA介导突触可塑性。我们有共免疫沉淀的darc1和copia来识别 潜在的相互作用者，我们将采用一种候选方法来找到遗传相互作用者。在初步工作中 我们发现需要DARC1才能在NMJ处进行适当的WNT途径信号传导。另外，我们观察到 Copia和Darc1结合了一些相同的蛋白质和mRNA，表明它们可能是 彼此对立，因此有可能解释它们在介导可塑性中看似相反的作用。 通过这笔赠款，我们将扩大我们对电动汽车贩运和突触可塑性的理解，而 描述了神经元通信中逆转录座子的新型物理功能。