Molecular Development of the Endolymphatic Duct and Sac

描述(摘自申请者摘要)：内耳内淋巴的动态平衡是 对内耳的感觉传导至关重要。未能维持 内淋巴内稳态被认为会导致耳聋、前庭功能障碍 和耳鸣的病理，如梅尼埃病或某些形式的 遗传性听力障碍。内淋巴管和内淋巴囊是关键。 维持这种液体动态平衡的结构。因此，关于 领域的分子发展是非常相关的。通过专注于一只鼠标 突变(Kreisler)，该应用程序试图定义 参与诱导和分化的分子途径。至 确定KR信令的早期目标，此应用程序将测试 假设原始区早期分子标记的表达是 与10-11天相比，纯合子KR胚胎中表达下调 控制。KR基因突变对人卵巢癌细胞分化的影响 发展中的领域将通过检验以下假设来研究： 胚胎第12天到第18天的细胞特异性基因组是 与对照组相比，Kreisler纯合子基因表达下调。为了方便 直接对正在开发的领域进行实验操作，该应用程序将 建立发育中的Kreisler耳囊和耳垂的体外模型。 实验将首先检验培养的克莱斯勒耳囊 在形态和功能上发育上模仿体内系统。这 然后将使用模型来检验病毒介导的表达的假设 KR能在体外挽救WIRS的表型。来检验这一假设 KR区的后脑来源诱导分化，我们将培养Kreisler 带有野生型后脑外植体的耳囊。这样的数据将提供洞察力 进入参与KR信号转导的分子通路，并参与 菲尔兹。PI对医学和科学的明显承诺已经得到证明 通过广泛追求临床和基础科学方面的培训 内耳生物学。这些努力的成功反映在他的 也证明了他有能力完成质量基础工作的出版物 科学调查。结合优秀的学术环境 在儿童医院研究基金会，RCA将允许PI 继续进行严格的科学培训，并成功解决具体问题 申请表中概述的目标。拟议的学习计划和科学 无疑将推动PI朝着他成功的目标前进 为未来的R01而竞争，并从长远来看，成为一名成功的 独立的临床医生科学家。值得注意的是，这项提议包括 具有挑战性但可实现的目标，将为 内耳发育领域。

DESCRIPTION (from applicant's abstract): Homeostasis of inner ear endolymph is critical to sensory transduction in the inner ear. Failure to maintain endolymph homeostasis is thought to result in deafness, vestibular dysfunction and tinnitus in pathologies such as Meniere's disease or certain forms of hereditary hearing impairment. The endolymphatic duct and sac (ELDS) are key structures in maintaining this fluid homeostasis. Therefore, data on the molecular development of the ELDS are very relevant. By focusing on a mouse mutant (kreisler) with an ELDS phenotype, this application seeks to define the molecular pathways involved in induction and differentiation of the ELDS. To determine early targets of kr signaling, this application will test the hypothesis that expression of early molecular markers of the ELDS anlage is down-regulated in homozygote kr embryos at embryonic day 10-11 compared to controls. The effects of kr mutation on cellular differentiation within the developing ELDS will be studied by testing the hypothesis that expression of a battery of genes specific for cells in the embryonic day 12 to 18 ELDS is down-regulated in kreisler homozygotes compared to controls. To facilitate direct experimental manipulation of the developing ELDS, this application will develop an in vitro model of the developing kreisler otocyst and ELDS. Experiments will first test the hypothesis that cultured kreisler otocysts developmentally mimic the in vivo system morphologically and functionally. This model will then be used to test the hypothesis that virally mediated expression of kr can rescue the ELDS phenotype in vitro. To test the hypothesis that hindbrain sources of kr induce ELDS differentiation, we will culture kreisler otocysts with wild-type hindbrain explants. Such data will provide insights into the molecular pathways involved in kr signaling and in development of the ELDS. The PI's obvious commitment to medicine and science has been demonstrated by his extensive pursuit of training in the clinical and basic science facets of inner ear biology. The success of these efforts are reflected in his publications which also demonstrate his ability to accomplish quality basic science investigation. In combination with the outstanding academic environment at Children's Hospital Research Foundation, the RCA will allow the PI to continue a rigorous scientific training and successfully address the specific aims outlined in the application. The proposed program of study and the science generated will undoubtedly advance the PI toward his goals of successfully competing for a future R01, and in the long term, becoming a successful independent Clinician scientist. Significantly, this proposal includes challenging but achievable goals that will provide important knowledge to the field of inner ear development.