Expanding the utility of Drosophila as a model for hearing research

描述(由申请人提供)：本提案中描述的研究的目标是促进我们对用于构建听觉和其他机械感觉系统的基本生物学机制的理解。这里提出的实验有两个目标。首先，我们计划开发一种普遍适用于从昆虫到哺乳动物的听觉系统研究的技术。其次，我们将利用这些技术来研究果蝇(果蝇)听觉器官的发育和分化。这些实验将扩大果蝇作为听力研究模型的用途，同时为了解果蝇听觉器官的分化和功能提供洞察力。实验的重点是CUT基因。我们已经证明，切割突变体是聋人，他们的耳聋与听觉器官机械转导结构的缺陷和神经退化有关。到目前为止，所使用的方法仅提供了切割功能的部分图景，对切割作用的具体机制知之甚少。在这里，我建议开发和利用一种创新的、强大的结合染色质免疫沉淀(ChIP)和微阵列表达分析的方法来识别受CUT调控的基因。CUT编码一种不寻常的同源结构域转录因子，其人类同源基因CCAAT-置换蛋白(CDP)/CUTL1参与细胞周期调节并发挥肿瘤抑制作用。拟议的实验将极大地扩展我们对Cut/CDP蛋白的功能以及它们是如何做到这一点的理解。将受到这些研究重大影响的领域包括：听力、机械感觉、切割/CDP生物学和转录因子生物学。鉴于果蝇和脊椎动物听觉器官发育之间正在出现的发育遗传相似之处，我预计拟议的实验产生的信息也将与人类的听力相关。

DESCRIPTION (provided by applicant): The goal of the research described in this proposal is to advance our understanding of fundamental biological mechanisms used to construct auditory and other mechanosensory systems. The experiments proposed here have two objectives. First, we plan to develop techniques that will be generally applicable to the investigation of auditory systems from insects to mammals. Second, we will use these techniques to study development and differentiation of the Drosophila (fruitfly) auditory organ. Together these experiments will expand the utility of Drosophila as a model for hearing research while providing insight into the differentiation and function of the Drosophila auditory organ. The experiments are focused the cut gene. We have shown that cut mutants are deaf and that their deafness is correlated both with defects in the mechanotransducing structures of the auditory organ and with neurodegeneration. The approaches used to date have provided only a partial picture of cut function with little insight into specific mechanisms of cut action. Here, I propose to develop and utilize an innovative and powerful combination of chromatin immunoprecipitation (ChIP) and microarray expression analysis to identify genes regulated by Cut. cut encodes an unusual homeodomain transcription factor whose human homolog, CCAAT-displacement protein (CDP)/CUTL1, has been implicated in cell cycle regulation and functions as a tumor suppressor. The proposed experiments will significantly expand our understanding of what Cut/CDP proteins do and how they do it. Fields that will be substantially impacted by these studies include: hearing, mechanosensation in general, Cut/CDP biology, and transcription factor biology. In light of the emerging developmental genetic parallels between Drosophila and vertebrate auditory organ development, I anticipate that information generated by the proposed experiments also will be relevant to human hearing.