Genetic Analysis of Inner Ear Development in Xenopus tropicalis

热带爪蟾内耳发育的遗传分析

• 批准号: 8307735
• 负责人: Richard M Harland
• 金额: $ 28.79万
• 依托单位: UNIVERSITY OF CALIFORNIA BERKELEY
• 依托单位国家: 美国
• 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/8307735
• 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

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）：听力或平衡缺陷很常见，由发育和环境原因引起。为了了解正常的内耳发育，我们将使用遗传上温顺的青蛙，热带爪蟾，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具有保守的特征，四足动物耳朵的发育与哺乳动物非常相似。耳朵发育的相关阶段可以在光学透明的蝌蚪中观察到，使这种动物成为耳朵突变体遗传筛选的理想选择。此外，耳发育缺陷会导致异常游泳行为和翻正反应丧失，因此也可以对解剖学上不明显的缺陷进行评分。作为对PA-06-365“听力和平衡的细胞谱系和发育研究”的回应，我们已经解决了“更全面地代表模型生物系统”以研究耳发育的需求。我们建议通过研究四足动物热带爪蟾（Xenopus tropicalis）的耳泡发育来促进我们对耳发育的理解，热带爪蟾是一种两栖动物，其内耳发育与哺乳动物非常保守。在一个小规模的遗传筛选使用非洲爪蟾热带，我们已经恢复突变体，破坏耳朵形态，耳石形成，和平衡/游泳行为。我们已经在其中两个影响耳锥发育和耳囊大小的基因中分离出受影响的基因。因此，我们已经证明筛选和恢复突变体、分析表型以及绘制和克隆受影响的基因是可能的。X. tropicalis与哺乳动物基因组显示出相当大的结构相似性和同线性，没有额外的全基因组重复的迹象，因此我们有信心我们可以鉴定出在哺乳动物中显示保守功能的各种基因中的隐性突变体。随着最近基因组组装和注释的改进，以及外显子捕获和高通量序列分析的技术进步，我们相信可以快速定位其他突变，并分离受影响的基因。在下一个资助期内，我们建议通过定位克隆来表征额外的突变等位基因，并将突变表型与潜在的分子和细胞缺陷联系起来。我们将联合收割机分子方法与经典的胚胎移植相结合，以了解突变体的细胞自主性，以及相互作用以产生功能性耳朵的信号和响应组织。