Studies on hair cell BK channels

毛细胞BK通道的研究

基本信息

批准号：
7860380
负责人：
DHASAKUMAR S NAVARATNAM
金额：
$ 35.33万
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 2012-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7860380
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 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 Location Measures Mechanics Membrane Potentials Methods Mining Modeling Molecular Mus N-Methyl-D-Aspartate Receptors Nature Neurons Oocytes Phosphorylation Physiological Processes Postdoctoral Fellow Potassium Channel Probability Property Proteins RNA Interference RNA Splicing Research Personnel Reverse Transcriptase Polymerase Chain Reaction Site Sorting - Cell Movement Source 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）：听力损失影响着大约 2500 万美国人。它还影响大约 35% 的 65 岁及以上老年人。该群体听力损失的主要原因是毛细胞损失。我们的兴趣是了解决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有助于开发出治疗毛细胞损失的合理疗法。毛细胞是声音的主要传感器，用于将声音的机械能转换为编码的神经元能量。在本提案中，我们试图了解大电导钾通道影响毛细胞功能的分子基础。具体来说，我们正试图阐明这些通道如何带来电调谐（一种频率辨别机制）的分子决定因素。我们还对确定它们如何与毛细胞基底外侧表面的电压门控 Ca 通道聚集和共定位的分子基础感兴趣。小鼠大电导钾通道的丧失会导致进行性听力损失。我们建议扩展先前已确定蛋白质一级结构变化的工作，现在寻求确定相互作用的蛋白质如何影响其功能。我们已经使用酵母 2 杂交技术从耳蜗中分离出了几种与该通道的有限部分相互作用的蛋白质。我们现在将确定这些相互作用的蛋白质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排序、聚类以及与电压门控 Ca 通道的共定位。此外，我们将扩展我们成功的酵母 2 杂交方法来识别与整个 BK 通道相互作用的其他蛋白质。这些蛋白质也将接受类似的检测。对于这些实验，我们将使用小鸡作为模型，并将使用我们开发的一组模块化测定法来测试它们在影响这些生理过程中的重要性。