Studies on hair cell BK channels

毛细胞BK通道的研究

基本信息

批准号：
7247111
负责人：
DHASAKUMAR S NAVARATNAM
金额：
$ 36.05万
依托单位：
YALE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2006
资助国家：
美国
起止时间：
2006-07-01 至 2011-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7247111
关键词：
Affect Alternative Splicing American Amino Acid Sequence Antibodies Binding Biolistics Biological Assay Buffers CDK5 gene Cell physiology Cells Chimeric Proteins Chinese Hamster Ovary Cell Cochlea Code Commercial Sources Cyclophosphamide/Fluorouracil/Prednisone Dependence Dickkopf-1 protein Discrimination Drosophila genus Equilibrium Event Family Fluorescence Resonance Energy Transfer Frequencies Goals Hair Cells Hearing Hybrids Immunoprecipitation Ion Channel Ions Kinetics Label Laboratories Localized Location Measures Mechanics Membrane Potentials Methods Mining Modeling Molecular Mus N-Methyl-D-Aspartate Receptors Nature Neurons Numbers Oocytes Phosphorylation Physiological Processes Postdoctoral Fellow Potassium Channel Probability Property Proteins RNA Interference RNA Splicing Range Rate Research Personnel Reverse Transcriptase Polymerase Chain Reaction Site Sorting - Cell Movement Structure Surface Synapses System Techniques Testing Time Transducers Turtles Ubiquitin Variant Vertigo Whole-Cell Recordings Work Yeasts base cell injury deafness gene cloning hearing impairment insight interest knock-down large-conductance calcium-activated potassium channels ligand gated channel mutant novel numb protein programs protein distribution research study sound sound frequency voltage yeast two hybrid system

项目摘要

DESCRIPTION (provided by applicant): Hearing loss affects approximately 25 Million Americans. It also affects approximately 35% of those 65 and older. The primary cause of hearing loss in this group is a result of a loss of hair cells. Our interest is in understanding the molecular events that determine hair cell function, and how these events are controlled. We anticipate that such an understanding will allow rational therapies to be developed to treat hair cell loss. Hair cells are the primary transducers of sound serving to convert the mechanical energy of sound to a coded a neuronal one. In this proposal we are seeking to understand the molecular basis of the large conductance potassium channel as it affects hair cell function. Specifically, we are trying to elucidate the molecular determinants of how these channels bring about electrical tuning, a mechanism of frequency discrimination. We are also interested in determining the molecular basis of how they are clustered and co-localized with the voltage gated Ca channel at the basolateral surface of hair cells. The loss of the large conductance potassium channel in mice results in progressive hearing loss. We propose to extend previous work that has identified variations in the primary structure of the protein, and now seek to determine how interacting proteins influence its function. We have already isolated several proteins from the cochlea that interact with a limited portion of this channel using the yeast 2 hybrid technique. We will now determine how these interacting proteins affect the kinetic properties of this channel. We will also determine how these proteins affect it basolateral sorting, clustering and co-localization with voltage gated Ca channels. In addition, we will extend our successful yeast 2 hybrid approach to identify other proteins that interact with entire BK channel. These proteins too will be subject to similar assays. For these experiments we wilt use the chick as a model and will use a modular set of assays that we have developed to test their significance in affecting these physiological processes.

描述（由申请人提供）：听力损失影响约2500万美国人。它也影响了65岁及以上的那些年龄段的35％。该组听力丧失的主要原因是毛细胞丧失的结果。我们的兴趣是了解确定毛细胞功能的分子事件以及如何控制这些事件。我们预计这种理解将允许发展理性疗法以治疗毛细胞损失。毛细胞是声音的主要传感器，可将声音的机械能转换为编码神经元的机械能。在此提案中，我们正在寻求了解大型电导钾通道的分子基础，因为它会影响毛细胞功能。具体而言，我们正在尝试阐明这些通道如何带来电气调整的分子决定因素，即频率歧视的机理。我们还有兴趣确定它们如何与毛细胞基底外侧表面的电压门控CA通道共凝聚和共定位的分子基础。小鼠大型电导钾通道的损失导致渐进的听力损失。我们建议扩大已经确定蛋白质主要结构变化的先前工作，现在试图确定相互作用蛋白如何影响其功能。我们已经使用酵母2混合技术从耳蜗中分离出几种蛋白质，这些蛋白质与该通道的有限部分相互作用。现在，我们将确定这些相互作用蛋白如何影响该通道的动力学特性。我们还将确定这些蛋白质如何影响其基底外侧分选，聚类和与电压门控CA通道共定位。此外，我们将扩展成功的酵母2混合方法，以识别与整个BK通道相互作用的其他蛋白质。这些蛋白质也将受到类似的测定。对于这些实验，我们将使用小鸡作为模型，并将使用一组模块化的测定方法来测试它们在影响这些生理过程中的重要性。