DYNAMIC INTERATION AMONG PROTEINS IN HAIR CELLS

描述（由申请人提供）：内耳中的Corti器官将声波产生的机械力转化为电信号，以通知中枢神经系统。为了完成这项任务，Corti器官中的毛细胞和支持细胞需要复杂而复杂的蛋白质网络来执行其功能。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耳蜗生理学的理解。这项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粘附蛋白。在AIM I中，我们建议研究CEACAM16在维持顶盖膜完整性和外毛细胞与顶盖膜之间的连接方面的功能作用。我们还将研究其与DFNA 4耳聋位点在体外和体内环境中的关系。Marshalin是一种微管负末端结合蛋白，我们将其鉴定为潜在的CDH23相关蛋白。在AIM II中，我们将研究Marshalin在Corti器官中的表达模式，并研究Marshalin在耳蜗发育过程中在毛细胞和支持细胞中的功能。我们将确定Marshalin与其潜在相关蛋白（包括CDH23、肌动蛋白、肌球蛋白7a和血影蛋白）之间相互作用的分子基础。这项研究将提供有关Marshalin在毛细胞和支持细胞分化过程中交联肌动蛋白丝和MT丝的作用以及Marshalin与最早确定的耳聋基因座DFNB 15的联系的急需信息。本研究也将提供更多关于CDH23在静纤毛形成中作用的信息。各种细胞，生物化学和电生理学方法，敲除和敲入小鼠模型，以及体内生理学方法将被用来追求我们的目标。从拟议的工作中得出的结论不仅可以识别新的耳聋基因，而且还将为进一步了解Corti器官的形成提供基础。这些知识对于理解许多与正常和无序通信过程的分子基础直接相关的基本生物学问题是非常必要的。

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.