GENES INVOLVED IN INNER EAR DEVELOPMENT

听力障碍是人类最常见的感觉障碍， 到80岁时，涉及多达50%的人口。多数 儿童严重听力障碍的病例涉及 内耳病理学内耳的病理学，包含听觉器官 轰动一时。内耳的正常发育和功能 取决于非常复杂的相互作用 不同类型的细胞群，因此预计将涉及许多 基因。这项提议的广泛目标是识别和变异 与小鼠发育和/或功能有关的基因 内耳以分子和遗传学的方式做出贡献 对这些复杂过程通常如何发生的描述。在……里面 此外，我们希望产生人类内耳的小鼠模型 功能障碍将有助于理解和潜在地 治疗潜在的病理。一种诱导基因陷阱筛选方法 被设计成同时标记和突变内源基因 插入LacZ报告基因的小鼠胚胎干细胞 提供了一种有效的方法来识别在 老鼠的内耳。老鼠的污渍是从 这些基因捕获细胞系，在一个案例中，LacZ插入 导致显性单侧听力损失。在他的应用中， 有人提议提炼和扩大该基因的体外部分。 通过改变诱导条件来捕获筛分。这些细胞系 在体外表达报告基因的人将被用来创造 嵌合胚胎将被检测报告基因 活动。基于报告基因选择的基因陷阱细胞系 发育中内耳的基因表达及分子鉴定 捕获的基因和捕获的基因的映射位置将是 用来产生老鼠星。捕获基因的表达 在正常的内耳发育过程中会伴随着 对基因陷阱杂合子进行β-半乳糖活性染色。承运商： 基因陷阱的插入将被交叉和分离 评估内耳功能和结构的变化 向量插入。最后，为了检验潜在的假设 捕获的基因在小鼠体内受到信号的调节 在培养的细胞中鉴定它们，表达TRAP基因 植入将在相关的胚胎中进行检查 信号通路受到遗传或药物的干扰 意思是。

Hearing impairment is the most common human sensory disorder, involving as much as 50% of the population by age 80. Most cases of profound hearing impairment in children involve pathology of the inner ear, which contains the organs of auditory sensation. The normal development and function of the inner ear depends upon complex interactions between an extraordinarily diverse group of cell types, and thus is expected to involve many genes. The broad aim of this proposal is to identify and mutate genes involved in the development and/or function of the mouse inner ear in order to contribute to the molecular and genetic description of how these complex processes normally occur. In addition, we hope to generate mouse models of human inner ear dysfunction that will be helpful in understanding and potentially treating the underlying pathology. An induction gene trap screen designed to simultaneously mark and mutate endogenous genes in mouse embryonic stem cells by insertion of a lacZ reporter gene has provided an effective means of identifying genes expressed in the mouse inner ear. Mouse stains have been developed from these gene traps cell lines, and in one case, the lacZ insertion gave rise to dominant unilateral hearing loss. In t his appliation, it is proposed to refine and expand the in vitro portion of the gene trap screen by altering the induction conditions. The cell lines that express the reporter gene in vitro will be used to create chimeric embroyos which will be examined for reporter gene activity. Gene trap cell lines selected on the basis of reporter gene expression in the developing inner ear, molecular identify of the trapped gene, and map position of the trapped gene, will be used to generate mouse starins. Expression of the trapped genes will be followed throughout normal inner ear development by staining gene trap heterozygotes for beta-gal activity. Carriers of the gene trap insertions will be intercrossed and the offsrping assessed for changes in inner ear funtion and structure attributable to the vector insertion. Finally, to test the underlying hypothesis that the trapped genes are regulated in mice by the signals used to identify them in cultured cells, expression of the gene trap insertions will be examined in embryos in which the relevant signaling pathway has been perturbed by genetic or pharmacologic means.