GENES INVOLVED IN INNER EAR DEVELOPMENT

听力障碍是人类最常见的感觉障碍， 涉及到80岁以上人口的50%。 最 儿童严重听觉障碍的个案包括 内耳病理学，包括听觉器官 感觉。 内耳的正常发育和功能 取决于一个非常复杂的相互作用 不同的细胞类型，因此预计将涉及许多 基因. 这项提案的主要目的是识别和变异 参与小鼠发育和/或功能的基因 内耳，以有助于分子和遗传 描述这些复杂过程通常是如何发生的。 在 此外，我们还希望建立人内耳的小鼠模型， 这将有助于理解和潜在的功能障碍， 治疗潜在的疾病 一种诱导基因陷阱筛选法， 旨在同时标记和突变内源基因， 通过插入lacZ报告基因的小鼠胚胎干细胞 提供了一种有效的方法来鉴定表达在 老鼠的内耳 老鼠斑是从 这些基因陷阱细胞系，在一个案例中，lacZ插入 导致了显性单侧听力损失 在他的申请中， 有人建议改进和扩大基因的体外部分， 通过改变诱导条件来捕获屏幕。 的细胞系 在体外表达报告基因的细胞， 嵌合胚胎，将检查报告基因 活动 基于报告基因选择的基因陷阱细胞系 内耳发育中的基因表达， 被捕获的基因和被捕获的基因的图谱位置将被 用于生成鼠标星号。 被困基因的表达 在正常内耳发育过程中， 对β-gal活性的基因陷阱杂合体染色。 携带者 基因陷阱插入物将被交叉， 评估内耳功能和结构的变化 到载体插入。 最后，为了验证基本假设， 被捕获的基因在老鼠体内受到信号的调节， 在培养的细胞中鉴定它们， 将在胚胎中检查插入，其中相关的 信号通路受到遗传或药理学的干扰， 手段

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.