EYA1 In Ear Development And Branchio-Oto-Renal Syndrome

EYA1 耳朵发育与鳃耳肾综合征

• 批准号: 7210704
• 负责人: JOSE M MANALIGOD
• 金额: $ 40.36万
• 依托单位: UNIVERSITY OF IOWA
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-04-15 至 2010-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7210704
• 关键词: Affect; Alternative Splicing; Animal Model; Attention; Auditory system; Branchio-Oto-Renal Syndrome; Child; Congenital Abnormality; Development; Diagnosis; Dominant-Negative Mutation; Ear; Embryo; Gene Expression; Gene Expression Alteration; Gene Mutation; Genes; Genetic; Genetic Transcription; Individual; Injection of therapeutic agent; Investigation; Labyrinth; Learning; Messenger RNA; Methods; Modeling; Molecular; Mutation; Oligonucleotides; Other Genetics; Pathogenesis; Pathway interactions; Pattern; Phenotype; Play; Protein Isoforms; Proteins; RNA Splicing; Reporting; Research; Research Personnel; Role; Sensory; Staging; Testing; Tissues; Transcript; Xenopus; Xenopus laevis; body system; deafness; hearing impairment; insight; mRNA Transcript Degradation; malformation; mutant; programs; protein expression; research study; tool

DESCRIPTION (provided by applicant): Congenital hearing loss affects ~ 1:1000 children, making it one of the most commonly diagnosed sensory birth defects. The long-term objective of this study is to use Xenopus laevis as a vertebrate model organism to investigate the genetic mechanisms of inner ear development and deafness. Targeted alteration of gene expression in Xenopus embryos, which has been successfully used in the investigation of other organ systems, is an ideal method to investigate the developmental dynamics of inner ear formation. We will initially focus on the role of EYA1 in inner ear development, since EYA1 mutations result in branchio-otorenal syndrome (BOR), a common syndromic forms of hearing loss with a unique phenotype that may be reproducible and easily recognizable in a model organism. This study has 3 specific aims: 1) to determine the spatial and temporal developmental expression patterns of EYA1 in the Xenopus laevis inner ear. A comparison will be made of the stage specific expression patterns of both EYA1 mRNA and protein in the inner ear, with specific attention to the presence and functional significance of alternative splicing in EYA1 expression. 2) To perform targeted EYA1 mRNA degradation with modified oligonucleotide injection of Xenopus embryos to determine its molecular and morphologic effects in the developing inner ear. If these experiments result in inner ear abnormalities comparable to that found in BOR, this would indicate haploin sufficiency as a likely mechanism for the development of the BOR phenotype. 3) To model the effects of EYA1 mRNA misexpression by embryo injection experiments involving excess normal message or altered mRNAs corresponding to mutations found in BOR. If inner ear malformations result from inappropriate EYA1 mRNA levels, this would indicate that dominant negative or abnormal protein activity underlies the dominant phenotype of BOR.

描述（由申请人提供）：先天性听力损失影响约1：1000的儿童，使其成为最常见的诊断感官出生缺陷之一。本研究的长期目标是利用非洲爪蟾作为脊椎动物模式生物，研究内耳发育和耳聋的遗传机制。在非洲爪蟾胚胎中靶向改变基因表达是研究内耳形成发育动力学的理想方法，已成功应用于其他器官系统的研究。我们最初将重点关注EYA 1在内耳发育中的作用，因为EYA 1突变会导致鳃耳肾综合征（BOR），这是一种常见的听力损失综合征形式，具有独特的表型，在模式生物中可能是可重复且易于识别的。本研究有3个具体的目的：1）确定EYA 1在非洲爪蟾内耳的时空发育表达模式。比较EYA 1 mRNA和蛋白质在内耳中的阶段特异性表达模式，特别注意EYA 1表达中选择性剪接的存在和功能意义。2)目的：应用修饰的寡核苷酸注射技术对非洲爪蟾胚胎进行靶向EYA 1 mRNA降解，以确定其对发育中内耳的分子和形态学影响。如果这些实验导致与BOR中发现的内耳异常相当的内耳异常，则这将表明单倍蛋白充足是BOR表型发展的可能机制。3)通过胚胎注射实验模拟EYA 1 mRNA错误表达的影响，该实验涉及过量的正常信息或与BOR中发现的突变相对应的改变的mRNA。如果内耳畸形是由EYA 1 mRNA水平不适当引起的，这将表明显性负性或异常蛋白活性是BOR显性表型的基础。