Elucidating the molecular mechanisms that shape the hair bundle

阐明塑造发束的分子机制

基本信息

批准号：
7440568
负责人：
Brian Michael McDermott
金额：
$ 43.02万
依托单位：
CASE WESTERN RESERVE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-06-05 至 2011-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7440568
关键词：
Actins Address Adult Apical BAIAP2 gene Behavioral Biological Biological Assay Biological Models Bundling CDC42 gene Cell physiology Cells Cellular biology Chimeric Proteins Collaborations Cytoskeletal Gene Cytoskeletal Proteins DNA DNA Microarray Chip Detection Development EPS8 gene Epithelial Cells Equilibrium Evaluation Excision Fellowship Functional disorder Gene Expression Profile Genes Goals Grant Hair Hair Cells Hearing Height Hepatocyte Homologous Gene Human Image Immunoelectron Microscopy In Situ Hybridization Inherited Ions Kinocilium Label Labyrinth Maintenance Maps Mechanoreceptors Methods Microarray Analysis Microtubules Molecular Morphogenesis Morphology Movement Mus Mutate Myosin ATPase National Institute on Deafness and Other Communication Disorders National Research Service Awards Oligonucleotide Microarrays Operative Surgical Procedures Optics Orthologous Gene Phase Play Population Positioning Attribute Postdoctoral Fellow Principal Investigator Proteins RNA RNA Cap-Binding Proteins Reverse Transcriptase Polymerase Chain Reaction Role Sensorineural Hearing Loss Shapes Staging Stereocilium Structure Surface System Techniques Testing Tissues Transcript Transcriptional Regulation Trimethoprim-Sulfamethoxazole Vertebrates Vesicle Water Movements Zebrafish Zebrafish Proteins actin 2 base deafness density design falls fascin functional genomics grasp in vivo insight interest kinetosome lateral line mutant positional cloning protein function public health relevance research study

项目摘要

DESCRIPTION (provided by applicant): An understanding of the morphogenesis and operation of hair cells requires identification of the genes expressed in these cells and of the functions of the cognate proteins. As an initial step towards this goal, we have used DNA-microarray technology to identify the transcripts present in hair cells of the adult zebrafish. After isolating pure populations of hair cells from the lagena of the inner ear, we amplified and labeled RNA and hybridized the resultant probes to oligonucleotide microarrays. To exclude genes expressed ubiquitously in epithelial cells, we conducted parallel experiments with hepatocytes and disregarded the genes that they expressed. The hair-cell transcriptome of the zebrafish includes genes involved in vesicle fusion, transcriptional regulation, and transmembrane ion movement; among these are homologs of genes that, when mutated, produce deafness in humans and mice. Additional genes encode proteins that are involved in cytoskeletal function, and might therefore contribute to the formation and maintenance of the hair bundle. These components include actin-binding and -capping proteins as well as constituents of the intraflagellar transport system. Finally, the transcriptional profile encompasses numerous genes of undetermined function. Using the molecular-biological techniques applicable to the zebrafish, we have begun to investigate the roles that some of these genes' products play in hair-bundle development. PUBLIC HEALTH RELEVANCE The hair cell is a mechanoreceptor of the inner ear and lateral-line system that is responsible for hearing, balance, and the detection of water movement by some aquatic vertebrates. The hair cell possesses a hair bundle, which holds the apparatus of mechanotransduction. To identify the protein constituents of the hair bundle and to give insights into hereditary deafness, we are developing a functional genomics approach in zebrafish.

描述（由申请人提供）：了解毛细胞的形态发生和运作需要鉴定这些细胞中表达的基因以及同源蛋白质的功能。作为实现这一目标的第一步，我们使用 DNA 微阵列技术来识别成年斑马鱼毛细胞中存在的转录本。从内耳耳膜中分离出纯毛细胞群后，我们扩增并标记 RNA，并将所得探针与寡核苷酸微阵列杂交。为了排除上皮细胞中普遍表达的基因，我们用肝细胞进行了平行实验，并忽略了它们表达的基因。斑马鱼的毛细胞转录组包括参与囊泡融合、转录调控和跨膜离子运动的基因；其中一些基因的同源物一旦发生突变，就会导致人类和小鼠耳聋。其他基因编码参与细胞骨架功能的蛋白质，因此可能有助于发束的形成和维持。这些成分包括肌动蛋白结合蛋白和加帽蛋白以及鞭毛内运输系统的成分。最后，转录谱包含许多功能未确定的基因。利用适用于斑马鱼的分子生物学技术，我们已经开始研究其中一些基因产物在发束发育中所起的作用。公共卫生相关性毛细胞是内耳和侧线系统的机械感受器，负责听力、平衡以及某些水生脊椎动物检测水的运动。毛细胞拥有一个毛束，其中包含机械传导装置。为了识别发束的蛋白质成分并深入了解遗传性耳聋，我们正在开发斑马鱼的功能基因组学方法。