Tip Link Regeneration in Auditory Hair Cells

描述（由申请人提供）：毛细胞尖端连接断裂和再生是低频噪声诱导的听力损失导致的暂时性阈值偏移的模型。这项研究将在分子水平上描述尖端连接再生的过程。该提案的目的是表征尖端链接蛋白和肌球蛋白马达的位置后，尖端链接中断，确定如何抑制尖端链接蛋白的相互作用会影响断裂的尖端链接的再生，并探索阵列的毛束蛋白是动态的尖端链接再生。该提案中的研究将检查毛束中蛋白质的免疫组织化学定位，通过扫描电子显微镜分析尖端链接的存在，测试截短的尖端链接蛋白在显性阴性实验中抑制再生的能力，并使用质谱法比较再生尖端链接的毛束的蛋白质含量。通过研究断裂的尖端连接的再生，将检查机械转导复合物响应于应力的动力学。相关性：大约有3600万美国成年人受到听力损失的影响。耳朵中检测声音的细胞很容易受到损害，这些细胞在这一点上不能被身体或科学方法所取代。这项研究的总体目标是了解这些细胞如何取代其功能所必需的受损成分，这对所有形式的听力损失都至关重要。

DESCRIPTION (provided by applicant): Hair cell tip link breakage and regeneration is a model for the temporary threshold shift resulting from low-frequency noise-induced hearing loss. The research in this proposal will characterize the process of tip link regeneration at a molecular level. The aims of this proposal are to characterize the location of tip link proteins and myosin motors after tip link disruption, determine how inhibiting the interaction of tip link proteins affects regeneration of broken tip links, and to explore the array of hair bundle proteins that are dynamic during tip link regeneration. The research in this proposal will examine immunohistochemical localization of proteins in the hair bundle, analyze the presence of tip links by scanning electron microscopy, test the ability of truncated tip link proteins to inhibit regeneration in dominant negative experiments, and compare the protein content of hair bundles that are regenerating tip links to control hair bundles using mass spectrometry. By studying regeneration of broken tip links, the dynamics of the mechanotransduction complex in response to stress will be examined. RELEVANCE: Approximately 36 million American adults are affected by hearing loss. Cells in the ear that detect sound are susceptible to damage, and these cells can not be replaced by the body or by scientific methods at this point. The overall aim of this research is to understand how these cells replace damaged components necessary for their function, which is of utmost importance to all forms of hearing loss.