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引起的听力损失引起的暂时性阈值漂移的模型。该方案中的研究将在分子水平上表征尖端连接再生的过程。该方案的目的是研究末端连接蛋白和肌球蛋白马达在末端连接中断后的位置，确定抑制末端连接蛋白之间的相互作用如何影响末端连接断裂的再生，并探索在末端连接再生过程中动态变化的发束蛋白阵列。这项建议中的研究将检测发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，在显性阴性实验中测试截短的尖端链接蛋白抑制再生的能力，并使用质谱仪比较再生尖端链接的发束的蛋白质含量以控制发束。通过研究断裂尖端环节的再生，我们将研究机械转导复合体对应力的反应动力学。相关性：大约3600万美国成年人受到听力损失的影响。耳朵中探测声音的细胞很容易受到损害，目前这些细胞不能被身体或科学方法取代。这项研究的总体目标是了解这些细胞如何取代其功能所需的受损成分，这对所有形式的听力损失都是至关重要的。

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.