Role of miRNAs in the development and function of a circuit for vocal learning

miRNA 在声音学习回路的发育和功能中的作用

• 批准号: 8803123
• 负责人: XIAOCHING LI
• 金额: $ 36.5万
• 依托单位: LSU HEALTH SCIENCES CENTER
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-12-01 至 2019-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8803123
• 关键词: 3' Untranslated Regions; Adolescent; Adult; Affect; Area; Behavioral; Brain; Broca' s area; Cell Nucleus; Cell Proliferation; Cerebellum; Corpus striatum structure; Dendritic Spines; Development; Diagnostic; Embryo; Environmental Risk Factor; Finches; Gene Expression; Genes; Genetic; Genetic study; Image; Impairment; In Vitro; Individual; Language; Lead; Learning; Mediating; Mental disorders; Messenger RNA; MicroRNAs; Molecular; Motor; Mutation; Neurons; Newborn Infant; Open Reading Frames; Outcome; Performance; Play; Procedures; Protein Biosynthesis; Recruitment Activity; Regulation; Regulator Genes; Research; Role; Site; Speech; Structure; Subfamily lentivirinae; Testing; Therapeutic Agents; Untranslated RNA; Ventricular; adult neurogenesis; autism spectrum disorder; base; density; developmental disease; dimer; in vivo; insight; lateral ventricle; learned behavior; mRNA Transcript Degradation; motor learning; nerve stem cell; nervous system development; nervous system disorder; neural circuit; neurogenesis; novel; novel diagnostics; paralogous gene; postnatal; public health relevance; relating to nervous system; research study; therapeutic target; transcription factor; vocal learning; young adult; zebra finch

 DESCRIPTION (provided by applicant): miRNAs are small non-coding RNA molecules that regulate gene expression by targeting the 3'-untranlated regions (UTRs) of mRNAs leading to mRNA degradation and suppression of protein synthesis. By coordinating and tuning gene expression, miRNAs play important roles in nervous system development and function. Dysregulation of miRNAs may lead to neural developmental disorders and mental illness. Here we propose to study two brain-enriched miRNAs, miR-9 and miR-139-5p, which potentially regulate the FOXP1 and FOXP2 genes. FOXP1 and FOXP2 are paralog transcription factors that function as a dimer and regulate the expression of numerous downstream genes important in neural circuit development. Mutations in the FOXP2 gene cause speech and language impairments, and both FOXP1 and FOXP2 have also been implicated in the Autism Spectrum Disorders (ASDs). An emerging hypothesis suggests that an adequate amount of functional FOXP2 is required for the proper development of the distributed neural circuitry for procedure learning and motor sequencing required for speech and language. However, regulation of the expression of FOXP1 and FOXP2 is not clear. Both the FOXP1 and FOXP2 genes have long 3'-UTRs (~4 kb), twice as long as their respective protein coding regions, suggesting that they are regulated by miRNAs. miR-9 and miR-139-5p are evolutionarily conserved and their targeting sites in the 3'-UTRs of FOXP1 and FOXP2 are also conserved, allowing us to study them in the zebra finch in the context of neural circuit development and vocal learning behavior. In the zebra finch brain, FOXP1 and FOXP2 are expressed in Area X, a nucleus in a cortical-striatal circuit required for vocal learning in juvenile zebra finches. Knockdown of FoxP2 in Area-X impairs vocal learning and reduces dendritic spine density in juvenile finches. Area X is largely composed of spiny neurons and a considerable number of new spiny neurons are generated in the germinal lining of the lateral ventricle adjacent to Area X and being recruited into Area X in young and adult zebra finches. We have shown that both miR-9 and miR-139-5p are expressed in Area X and in the ventricular zone. miR-9 is also known to regulate neural progenitor proliferation and differentiation during embryonic brain development. These collective observations led us to hypothesize that miR-9 and miR-139-5p regulate FOXP1 and FOXP2, and play important roles in regulating postnatal neurogenesis, neuronal dendritic development, and vocal learning behavior. This hypothesis will be tested using a combination of molecular, cellular, and behavioral approaches. First, we will test whether miR-9 and miR-139-5p regulate FoxP1 and FoxP2 in vitro; we will also examine how the expression of miR-139-5p and FoxP1 is regulated in the zebra finch brain during vocal development and in adults (we already studied miR-9 and FoxP2). Then we will use lentiviruses to manipulate the expression of miR-9 and miR-139-5p in Area X and test the effects on vocal learning and performance. We will also study the roles of miR-9 and miR-139-5p in regulating neurogenesis in postnatal and adult Area X, and study the roles of miR-9 and miR-139-5p in regulating dendritic structures in spiny neurons in Area X. This research promises to establish novel post-transcriptional regulatory relationships between miR-9/miR- 139-5p and FOXP1/FOXP2, and to delineate the roles of miR-9 and miR-139-5p in regulating neurogenesis and dendritic development in a neural circuit important for vocal learning behavior. As the striatum is at the center of many mental illness and neurological disorders, these results may open new opportunities to study how genetic and environmental factors contribute to these conditions via miRNA-mediated gene regulatory networks. With further research, these miRNAs may be developed into diagnostic markers or therapeutic agents.

 描述(申请人提供)：miRNAs是一种小的非编码RNA分子，通过靶向mRNAs的3‘-非翻译区(UTRs)来调节基因表达，导致mRNA降解和蛋白质合成受到抑制。MiRNAs通过协调和调节基因表达，在神经系统发育和功能中发挥重要作用。MiRNAs的失调可能导致神经发育障碍和精神疾病。在这里，我们建议研究两个大脑丰富的miRNAs，miR-9和miR-139-5p，它们可能调节Foxp1和FOXP2基因。Foxp1和FOXP2是平行转录因子，起二聚体的作用，调节许多在神经回路发育中重要的下游基因的表达。FOXP2基因的突变会导致言语和语言障碍，Foxp1和FOXP2也都与自闭症谱系障碍(ASD)有关。一种新的假说表明，对于程序学习和语言所需的运动序列的分布式神经回路的适当发展，需要足够数量的功能性FOXP2。然而，Foxp1和FOXP2的表达调控尚不清楚。Foxp1和FOXP2基因都有较长的3‘-UTRs(~4kb)，是各自蛋白编码区的两倍，表明它们受miRNAs调控。MiR-9和miR-139-5p在进化上是保守的，它们在Foxp1和FOXP2的3‘-UTRs中的靶点也是保守的，这使得我们可以在斑雀神经回路发育和发声学习行为的背景下对它们进行研究。在斑马雀的大脑中，Foxp1和FOXP2在X区表达，X区是幼年斑雀发声学习所需的皮质-纹状体回路中的一个核。在X区敲除FoxP2会损害幼雀的发声学习，并降低树突棘的密度。X区主要由棘神经元组成，幼龄和成年斑马雀在毗邻X区的侧脑室生发衬里产生相当数量的新棘神经元，并被招募到X区。我们发现miR-9和miR-139-5p在X区和脑室带均有表达。MIR-9还被认为在胚胎大脑发育过程中调节神经前体细胞的增殖和分化。这些集体观察使我们假设miR-9和miR-139-5p调节Foxp1和FOXP2，并在调节出生后神经发生、神经元树突发育和发声学习行为中发挥重要作用。这一假设将使用分子、细胞和行为方法的组合进行验证。首先，我们将测试miR-9和miR-139-5p是否在体外调节FoxP1和FoxP2；我们还将检测miR-139-5p和FoxP1在斑雀发声发育过程中和成年斑雀脑中的表达是如何调节的(我们已经研究了miR-9和FoxP2)。然后，我们将使用慢病毒来操纵X区miR-9和miR-139-5p的表达，并测试其对发声学习和表演的影响。我们还将研究miR-9和miR-139-5p在出生后和成年X区神经发生调控中的作用，以及miR-9和miR-139-5p在X区棘神经元树突结构调控中的作用。本研究旨在建立miR-9/miR-139-5p和Foxp1/FOXP2之间新的转录后调控关系，并阐明miR-9和miR-139-5p在调节发声学习行为重要神经回路中的神经发生和树突发育中的作用。由于纹状体是许多精神疾病和神经疾病的中心，这些结果可能为研究遗传和环境因素如何通过miRNA介导的基因调控网络导致这些疾病提供新的机会。随着进一步的研究，这些miRNAs可能被开发成诊断标记物或治疗剂。