Genetic Analysis of Inner Ear Development in Xenopus tropicalis

热带爪蟾内耳发育的遗传分析

基本信息

批准号：
8509655
负责人：
Richard M Harland
金额：
$ 27.3万
依托单位：
UNIVERSITY OF CALIFORNIA BERKELEY
依托单位国家：
美国
项目类别：
财政年份：
2011
资助国家：
美国
起止时间：
2011-08-01 至 2016-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8509655
关键词：
ATP-Binding Cassette Transporters Acceleration Adaptor Signaling Protein Address Affect Aging Alleles Amphibia Animal Model Animals Auditory Behavior Behavioral Candidate Disease Gene Cell Lineage Cells Chemicals Clinical Data Defect Development Disease Ear Embryology Equilibrium Esthesia Event Exons Family Fishes Functional disorder Future Genes Genetic Genetic Screening Genome Grant Hearing Human Individual Inherited Labyrinth Lead Lesion Link Mammals Maps Melanophores Methods Molecular Morphogenesis Morphology Multi-Drug Resistance Mus Mutagenesis Mutant Strains Mice Mutation Nature Noise Nonsense Codon Otic Placodes Otic Vesicle Pathway interactions Pattern Pharmaceutical Preparations Phenotype Pigmentation physiologic function Process Proteins Rana Research Resolution Role Sensory Sequence Analysis Signal Transduction Sorting - Cell Movement Staging Structure Swimming Synteny System Tadpoles Tissues Transplantation Vertebrates Vesicle Xenopus base comparative deafness equilibration disorder gene function genetic analysis hearing impairment high risk insight mammalian genome member mutant novel otoconia positional cloning response

项目摘要

DESCRIPTION (provided by applicant): Deficits in hearing or balance are common, and result from both developmental and environmental causes. In order to understand normal inner ear development, we will use the genetically tractable frog, Xenopus tropicalis, to investigate the genetic network underlying ear development. The structure and development of the inner ear shows conserved features among the vertebrates, and the development of the ear of the tetrapod is very similar to that of mammals. The relevant stages of ear development can be observed in the optically clear tadpole, making this animal ideal for genetic screens for ear mutants. Furthermore, defects in ear development lead to abnormal swimming behavior and loss of the righting response, so that defects that may not be anatomically obvious can also be scored. In response to PA-06-365, "Cell Lineage and Developmental Studies in Hearing and Balance" we have addressed the stated need for "more comprehensive representation of model organisms systems" to study ear development. We propose to advance our understanding of ear development by examining otic vesicle development in the tetrapod Xenopus tropicalis, an amphibian whose inner ear development is well-conserved with mammals. In a small-scale forward genetic screen using Xenopus tropicalis we have recovered mutants that disrupt ear morphology, otolith formation, and balancing/swimming behavior. We have isolated the affected gene in two of these that affect otoconial development and otocyst size. Thus we have shown that it is possible to screen for and recover mutants, analyze the phenotype, and map and clone the affected genes. The genome of X. tropicalis shows considerable structural similarity and synteny with mammalian genomes, with no sign of additional whole genome duplications, so we are confident that we can identify recessive mutants in a variety of genes which show conserved functions with the mammals. With recent improvements in the genome assembly and annotation, and technical advances in exon capture and high throughput sequence analysis, we are confident that other mutations can be rapidly mapped, and the affected genes isolated. In the next grant period, we propose to characterize additional mutant alleles by positional cloning, and link mutant phenotypes with the underlying molecular and cellular defects. We combine molecular approaches with classical embryological transplantations to understand the cel autonomy of mutants, and the signaling and responding tissues that interact to produce the functioning ear.

描述（由申请人提供）：听力或平衡的缺陷很常见，并且是由于发展和环境原因而造成的。为了理解正常的内耳发育，我们将使用可遗传牵引的青蛙Tropicalis进行研究，以研究耳朵发育的遗传网络。内耳的结构和发育在脊椎动物中显示出保守的特征，而四足动物耳朵的发育与哺乳动物的发育非常相似。可以在光学透明的tpole中观察到耳朵发育的相关阶段，这使该动物非常适合耳朵突变体的遗传筛选。此外，耳朵发育的缺陷导致游泳行为异常和矫正反应的丧失，因此在解剖学上可能并不明显的缺陷也可以得分。为了回应PA-06-365，“听力和平衡中的细胞谱系和发展研究”，我们已经解决了对“模型生物系统更全面表示”以研究耳朵发育的必要性。我们建议通过检查Tetrapod Xenopus tropicalis的耳囊发育，这是一种两栖动物，这是一种两栖动物，其内耳发育与哺乳动物的内部发育良好。在使用Tropicalis Xenopus的小型前向遗传筛选中，我们恢复了破坏耳朵形态，耳石形成和平衡/游泳行为的突变体。我们已经在影响耳鼻喉发育和耳囊大小的两个中分离了受影响的基因。因此，我们已经表明，可以筛选和恢复突变体，分析表型，并绘制并克隆受影响的基因。 X. topicalis的基因组显示出相当大的结构相似性和与哺乳动物基因组的同步，没有额外的整个基因组重复的迹象，因此我们有信心我们可以鉴定出各种基因中显示出与哺乳动物保持保守功能的隐性突变体。随着基因组组装和注释的最新改进，以及外显子捕获和高吞吐量序列分析的技术进步，我们相信可以快速映射其他突变，并且受影响的基因隔离。在下一个赠款期间，我们建议通过位置克隆来表征其他突变等位基因，并将突变表型与潜在的分子和细胞缺陷联系起来。我们将分子方法与经典的胚胎移植相结合，以了解突变体的CEL自主权，以及相互作用以产生功能性耳朵的信号传导和反应组织。