PAX-2 AND K-CADHERIN AND INNER EAR DEVELOPMENT

PAX-2 和 K-钙粘蛋白与内耳发育

• 批准号: 2518048
• 负责人: JANI E LEWIS
• 金额: $ 2.22万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-09-01 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/2518048
• 关键词: antisense nucleic acid; cadherins; chick embryo; developmental genetics; developmental neurobiology; ganglions; gene induction /repression; innervation; labyrinth; neurogenetics; organ culture; otocyst /otolith; transcription factor

The long term goals of this proposal are to examine the molecular and cellular bases of very early events in inner ear development in order to understand the mechanisms that give rise to congenital deafness. The specific aims of this proposal focus on examining the importance of the developmentally regulated transcription factor, Pax-2, one of the earliest genes expressed in the otic placode and its potential downstream target K-cadherin (cCad6B in the chick). The first specific aim is designed to set up a blueprint of the mRNA and protein expression patterns of Pax-2 and K-cadherin from otic placode invagination (stage 8) through development of the three dimensional inner ear structure (stage 38). The second and third specific aims further examine the correlation between Pax-2 and K-cadherin expression. In both aims the localization of Pax2 and K-cadherin mRNA and protein will be examined in otocysts which have been either surgically manipulated (in specific aim 2) or exposed to factors causing overexpression or downregulation of the Pax-2 gene product (in specific aim 3). Both these sets of experiments will be performed in ovo and are expected to reveal the important interactions between the cellular environment and Pax-2 expression in addition to looking at the correlation between Pax-2 and K-cadherin expression. The fourth specific aim examines the importance of K-cadherin function for formation of the statoacoustic ganglion. These experiments involve functional blocking of K-cadherin activity and will be performed both in vitro, on organ cultures, and in vivo.

该提案的长期目标是检查分子和 内耳发育中非常早期事件的细胞基础，以便 了解引起先天性耳聋的机制。这 该提案的具体目标侧重于审查 发育调节转录因子 Pax-2，其中之一 耳基板中表达的最早基因及其潜在的下游 目标 K-钙粘蛋白（小鸡中的 cCad6B）。第一个具体目标是 旨在建立 mRNA 和蛋白质表达的蓝图 耳基板内陷中 Pax-2 和 K-cadherin 的模式（阶段 8）通过三维内耳结构的发展 （第 38 阶段）。第二个和第三个具体目标进一步审查了 Pax-2 和 K-钙粘蛋白表达之间的相关性。在这两个目标中 Pax2 和 K-cadherin mRNA 和蛋白质的定位将在 经过手术操作的耳囊（特定目的 2）或暴露于导致过度表达或下调的因素 Pax-2 基因产物（具体目标 3）。这两组实验 将在 ovo 中进行，预计将揭示重要的 细胞环境与 Pax-2 表达之间的相互作用 除了查看 Pax-2 和 K-钙粘蛋白之间的相关性之外 表达。第四个具体目标考察 K-钙粘蛋白的重要性 静声神经节的形成的功能。这些实验 涉及 K-钙粘蛋白活性的功能性阻断，并将进行 体外、器官培养和体内。