DYNAMIC INTERATION AMONG PROTEINS IN HAIR CELLS

描述(申请人提供)：内耳的Corti器官将声波产生的机械力转化为电信号，通知中枢神经系统。为了完成这一任务，Corti器官中的毛细胞和支持细胞需要复杂和复杂的蛋白质网络来执行它们的功能。发现单个蛋白质及其在Corti器官蛋白质网络中的作用将丰富我们对正常和受损的耳蜗生理学的理解。这项拟议工作的目标是确定两种新发现的蛋白质的功能：CEACAM16和Marshalin。CEACAM16是我们在耳蜗中发现的一种细胞黏附蛋白。在AIM I中，我们建议研究CEACAM16在维持被膜完整性以及外毛细胞与被膜连接中的功能作用。我们还将在体外和体内环境中研究其与DFNA4耳聋基因座的关系。Marshalin是一种微管负端结合蛋白，我们发现它是一种潜在的CDH23相关蛋白。在AIM II中，我们将研究马沙林在Corti器官中的表达模式，并研究马沙林在耳蜗发育过程中毛细胞和支持细胞的功能。我们将确定Marshalin与其潜在相关蛋白之间相互作用的分子基础，包括CDH23、肌动蛋白、肌球蛋白7a和血影蛋白。这项研究将提供马沙林在毛细胞和支持细胞分化过程中交联肌动蛋白细丝和MT细丝中的作用，以及马沙林与耳聋基因座DFNB15的联系，DFNB15是最早发现的耳聋基因座之一。这项研究还将提供有关CDH23在立体纤毛形成中的作用的进一步信息。将使用各种细胞、生化和电生理方法、基因敲除和敲除小鼠模型以及体内生理学方法来追求我们的目标。从拟议的工作中得出的结论不仅将使可能识别新的耳聋基因，还将为进一步了解Corti器官的形成提供基础。要理解许多与正常和无序通讯过程的分子基础直接相关的基本生物学问题，这些知识是非常必要的。

DESCRIPTION (provided by applicant): The organ of Corti in the inner ear converts the mechanical force arising from sound waves into electrical signals to inform the central nervous system. To accomplish this task, hair cells and supporting cells in the organ of Corti need complex and sophisticated protein networks to perform their functions. Discovering the individual proteins and their roles in the protein networks of the organ of Corti will enrich our understanding of normal and impaired cochlear physiology. The goal of the proposed work is to determine functions of two newly identified proteins: CEACAM16 and Marshalin. CEACAM16 is a cell adhesion protein that we discovered in the cochlea. In AIM I, we propose to investigate functional roles of CEACAM16 for maintaining the tectorial membrane integrity and the connections between outer hair cells and the tectorial membrane. We will also investigate its relation to the DFNA4 deafness locus in both in vitro and in vivo environments. Marshalin is a microtubule-minus-end binding protein that we identified as a potential CDH23-associated protein. In AIM II, we will examine Marshalin expression patterns in the organ of Corti and investigate Marshalin's function in both hair cells and supporting cells during cochlear development. We'll determine the molecular basis of interactions between Marshalin and its potential associated proteins including CDH23, actin, myosin 7a and spectrin. The investigation will provide much-needed information regarding Marshalin's roles in cross-linking actin filaments and MT filaments during hair cells and supporting cells differentiation, as well as Marshalin's connection with deafness locus DFNB15, one of earliest identified deafness loci. The investigation will also offer further information regarding CDH23 role for stereocilia formation. Various cellular, biochemical and electrophysiological methods, knockout and knockin mouse models, as well as in vivo physiology methods will be used to pursue our goals. The conclusions derived from the proposed work will not only allow the possible identification of new deafness genes, but will also provide the foundation for further understanding of formation of the organ of Corti. Such knowledge is sorely needed to understand many fundamental biological questions that are directly relevant to the molecular bases of normal and disordered communication processes.