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: 9199444
• 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/9199444
• 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; Dimerization; Embryo; Environmental Risk Factor; FOXP1 gene; FOXP2 gene; Finches; Gene Expression; Genes; Genetic; Genetic Transcription; Genetic study; 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; diagnostic biomarker; dimer; experimental study; imaging study; in vivo; insight; knock-down; language impairment; lateral ventricle; learned behavior; mRNA Transcript Degradation; 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; 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（由申请人提供）：miRNA是小的非编码RNA分子，其通过靶向mRNA的3 '-非翻译区（UTR）调节基因表达，导致mRNA降解和抑制蛋白质合成。通过协调和调节基因表达，miRNAs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，我们建议研究两种脑富集的miRNAs，miR-9和miR-139- 5 p，它们可能调节FOXP 1和FOXP 2基因。FOXP 1和FOXP 2是作为二聚体起作用的旁转录因子，并调节在神经回路发育中重要的许多下游基因的表达。FOXP 2基因的突变会导致言语和语言障碍，FOXP 1和FOXP 2也与自闭症谱系障碍（ASD）有关。一个新兴的假说表明，需要足够数量的功能FOXP 2的适当发展的分布式神经回路的程序学习和运动所需的语音和语言的排序。然而，FOXP 1和FOXP 2的表达调控尚不清楚。FOXP 1和FOXP 2基因都有很长的3 '-UTR（~4 kb），是它们各自蛋白编码区的两倍长，这表明它们受miRNA调控。miR-9和miR-139- 5 p在进化上是保守的，它们在FOXP 1和FOXP 2的3 '-UTR中的靶位点也是保守的，这使得我们能够在斑胸草雀的神经回路发育和发声学习行为的背景下研究它们。在斑胸草雀的大脑中，FOXP 1和FOXP 2在X区表达，这是幼年斑胸草雀发声学习所需的皮质-纹状体回路中的一个核团。在Area-X中FoxP 2的敲低损害了声音学习并降低了幼年雀的树突棘密度。X区主要由多刺神经元组成，并且相当数量的新的多刺神经元产生于与X区相邻的侧脑室的生发层中，并且在年轻和成年斑胸草雀中被招募到X区。我们已经证明miR-9和miR-139- 5 p都在X区和心室区表达。还已知miR-9在胚胎脑发育期间调节神经祖细胞增殖和分化。这些集体观察使我们假设miR-9和miR-139- 5 p调节FOXP 1和FOXP 2，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测试。首先，我们将测试miR-9和miR-139- 5 p是否在体外调节FoxP 1和FoxP 2;我们还将研究miR-139- 5 p和FoxP 1的表达在斑胸草雀大脑中如何在发声发育过程中和成年人中进行调节（我们已经研究了miR-9和FoxP 2）。然后，我们将使用慢病毒操纵X区中miR-9和miR-139- 5 p的表达，并测试其对发声学习和表现的影响。我们还将研究miR-9和miR-139- 5 p在调节出生后和成年X区神经发生中的作用，并研究miR-9和miR-139- 5 p在调节X区棘神经元树突结构中的作用。本研究有望建立miR-9/miR- 139- 5 p和FOXP 1/FOXP 2之间新的转录后调控关系，并阐明miR-9和miR-139- 5 p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系统疾病的中心，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促进这些疾病提供新的机会。随着研究的深入，这些miRNAs有可能被开发成诊断标志物或治疗药物。