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：
8258910
负责人：
Brian Michael McDermott
金额：
$ 41.2万
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 2014-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8258910
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 Health 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 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 operation positional cloning protein function 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）：对毛细胞的形态发生和操作的理解需要鉴定这些细胞中表达的基因和同源蛋白的功能。作为朝着这一目标的第一步，我们使用了DNA微阵列技术来识别成年斑马鱼毛细胞中存在的成绩单。在从内耳的拉格纳分离出纯毛细胞群后，我们放大并标记了RNA，并将所得探针杂交到寡核苷酸微阵列中。为了排除在上皮细胞中普遍表达的基因，我们用肝细胞进行了平行的实验，并无视它们表达的基因。斑马鱼的发胶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。其中是基因的同源物，当突变时会在人类和小鼠中产生耳聋。其他基因编码参与细胞骨架功能的蛋白质，因此可能有助于形成和维持毛发束。这些成分包括肌动蛋白结合和 - 盖蛋白以及flagellar运输系统的成分。最后，转录曲线包含许多不确定函数的基因。使用适用于斑马鱼的分子生物学技术，我们开始研究这些基因产品中一些产品在脱发发育中起的作用。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，平衡和某些水生脊椎动物对水的运动检测。毛细胞具有一个头发束，可容纳机械转导的设备。为了确定头发束的蛋白质成分并为遗传性耳聋提供见解，我们正在斑马鱼中开发一种功能性基因组学方法。