MicroRNA function in ascidian sensory neurons

海鞘感觉神经元中的 MicroRNA 功能

• 批准号: 8889247
• 负责人: ROBERT W ZELLER
• 金额: $ 18.56万
• 依托单位: SAN DIEGO STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-07-10 至 2017-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8889247
• 关键词: Address; Afferent Neurons; Algorithms; Animal Model; Animals; Biological Assay; Blast Cell; Child; Childhood; Chordata; Cilia; Complex; Defect; Development; Disease; Embryo; Equilibrium; Gene Cluster; Gene Expression Profile; Gene Expression Regulation; Gene Targeting; Genes; Genome; Genomics; Hair Cells; Health; Hearing; Hearing Impaired Persons; Human; In Situ Hybridization; Knowledge; Labyrinth; Larva; Lead; Left; Maintenance; Measures; MicroRNAs; Modeling; Molecular; Morphogenesis; Mus; Mutate; Mutation; Neurons; Normal Cell; Nucleic Acid Probes; Organ; Pattern; Peripheral Nervous System; Play; Point Mutation; Population; Porifera; Process; Processed Genes; Regulation; Regulator Genes; Reporter; Reporter Genes; Research; Role; Running; Seeds; Sensory; Sensory Hair; Sister; Site; Tail; Tissues; Transgenes; Transgenic Organisms; United States; Untranslated Regions; Urochordata; Vertebrates; ascidian; cell motility; cell type; comparative; deafness; hearing impairment; human disease; in vivo; insight; intercellular communication; locked nucleic acid; marine organism; member; neuron development; novel; response; sensor; transcription factor

DESCRIPTION (provided by applicant): The inner ear is a complex organ that provides us with the ability to hear and maintain balance. The development and morphogenesis of the inner ear depends upon precise temporal and spatial gene expression patterns, cell signaling and cell migration. Disruption of this regulation can lead to loss of hearing, which is common in the human population. Two or three out of every 1,000 children in the United States are born deaf or hard-of-hearing. More lose their hearing later during childhood and approximately one-third of people older than 60, and half older than 85, have hearing loss. Sensory neurons in the inner ear express a variety of microRNAs (miRs), including a three miR cluster comprised of miR-96, miR-182 and miR-183. Mutations in the seed sequence of miR-96 have been shown to result in hearing defects. A small number of inner ear target genes have been identified; however our knowledge on the overall role of these miRs and sensory neuron specification, development and maintenance is largely unknown. We have found that the sensory neurons of the ascidian larva express a transgene reporter for the ascidian miR-96, miR-182 and miR-183 cluster. Because animal sensory neurons are derived from a common ancestral cell type, ascidian sensory neurons likely share numerous sensory miR target genes with vertebrates. Ascidians are the sister group of vertebrates, but have small genomes and simplified gene regulatory networks making them ideal for regulatory analysis. We have developed the ascidian embryo as a platform for identifying and characterizing microRNAs and their gene targets. Here we will characterize the miR-96, -182 and -183 cluster and investigate conserved and unique target genes in ascidian embryos. The Specific Aims of this proposal are to: 1) Characterize expression of the sensory miR cluster in Ciona embryos. In situ hybridization with locked nucleic acid (LNA) probes, miR sponges, and ectopic transcription factor expression will be used to examine miR expression. 2) Identify gene targets regulated by the sensory miR cluster. Sensory miR gene targets shared between ascidians and humans will be assayed in vivo using a transgenic sensory assay. This research addresses fundamental questions of miR target gene regulation and will help promote ascidians as a model organism for comparative regulatory studies not only as related to hearing loss, but to other diseases as well. In addition, this research may provide novel potential targets for the treatment of sensory neuronal diseases, i.e. deafness, which are associated with SMC targets.

描述（由申请人提供）：内耳是一个复杂的器官，它为我们提供了听到和维持平衡的能力。内耳的发育和形态发生取决于精确的时间和空间基因表达模式，细胞信号传导和细胞迁移。这种法规的破坏可能导致听力丧失，这在人口中很常见。在美国，每1000名儿童中有两个或三个出生于聋哑人或听力障碍。在童年后期晚些时候，有更多的听力，大约三分之一的年龄超过60岁，年龄在85岁以上的人有听力损失。内耳中的感觉神经元表达各种microRNA（miR），包括由miR-96，miR-182和miR-183组成的三个miR簇。 miR-96种子序列中的突变已显示出导致听力缺陷。已经确定了少量的内耳靶基因。但是，我们对这些miR和感觉神经元规范，开发和维护的总体作用的了解在很大程度上是未知的。我们发现，海囊幼虫的感觉神经元表达了海鞘miR-96，miR-182和miR-183簇的转基因记者。由于动物感觉神经元来自普通的祖传细胞类型，因此海囊感觉神经元可能与脊椎动物共享许多感觉miR靶基因。海沿岸是脊椎动物的姊妹组，但具有较小的基因组和简化的基因调节网络，使其非常适合调节分析。我们已经开发了海囊胚作为识别和表征microRNA及其基因靶标的平台。在这里，我们将表征miR -96，-182和-183群集，并研究海囊胚胎中保守和独特的靶基因。该建议的具体目的是：1）表征ciona胚胎中感觉miR簇的表达。与锁定的核酸（LNA）探针，miR海绵和异位转录因子表达的原位杂交将用于检查miR表达。 2）确定受感觉miR簇调节的基因靶标。感觉miR基因靶标在体内使用转基因感觉测定法分析了海沿岸和人类之间的靶标。这项研究解决了MIR靶基因调节的基本问题，并将有助于促进海沿岸作为模型生物体，不仅与听力损失有关，而且与其他疾病有关。此外，这项研究可能为治疗感觉神经元疾病的治疗（即与SMC靶标有关的耳聋）提供新的潜在靶标。