Molecular Development of the Endolymphatic Duct and Sac

描述（来自申请人摘要）：内耳内淋巴的稳态是 对内耳的感觉传导至关重要。 未能遵守本 内淋巴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 内淋巴管和囊（ELDS）是关键 维持这种液体平衡的结构。 因此， ELDS的分子发展是非常相关的。 通过关注一只老鼠 突变体（kreisler），本申请试图定义具有ELDS表型的 参与ELDS诱导和分化的分子途径。 到 确定kr信号的早期目标，此应用程序将测试 假设ELDS原基早期分子标记的表达 在胚胎第10-11天的纯合子kr胚胎中下调， 对照 kr基因突变对人乳腺癌细胞分化的影响 发展ELDS将通过测试的假设，即表达一个 一组对胚胎第12至18天的ELDS细胞特异的基因， 与对照相比，在Kreisler纯合子中下调。 以促进 直接实验操作的发展ELDS，这种应用将 建立发育中的Kreisler耳囊和ELDS的体外模型。 实验将首先检验培养的克氏耳囊 在形态和功能上发育模仿体内系统。 这 然后将使用模型来检验病毒介导的表达 kr在体外可挽救ELDS表型。 为了验证这个假设， kr的后脑来源诱导ELDS分化，我们将培养kreisler 耳囊肿与野生型后脑外植体。 这些数据将提供洞察力 进入参与kr信号传导的分子途径， ELDS。 PI对医学和科学的明显承诺已经被证明 通过他在临床和基础科学方面的广泛追求， 内耳生物学 这些努力的成功反映在他的 出版物，也证明了他的能力，完成质量的基本 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 继续严格的科学训练，并成功地解决具体的 在申请中列出的目标。 拟议的研究计划和科学 毫无疑问，这将使PI朝着成功的目标前进。 竞争未来的R 01，从长远来看，成为一个成功的 独立临床科学家。 值得注意的是，该提案包括 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.