Hearing loss in claudin 11-null mice

密蛋白 11 缺失小鼠的听力损失

• 批准号: 6673131
• 负责人: Alexander Gow
• 金额: $ 24.05万
• 依托单位: WAYNE STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-07-15 至 2008-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6673131
• 关键词: auditory pathways; beta galactosidase; cochlea; complementary DNA; congenital ear disorder; electron microscopy; endolymph; gene expression; genetically modified animals; hearing disorders; histology; immunocytochemistry; in situ hybridization; laboratory mouse; labyrinth; membrane proteins; northern blottings; organ of Corti; polymerase chain reaction; potassium; tight junctions; vestibular apparatus; western blottings

DESCRIPTION (provided by applicant): The long term goals of this project are to define the functions of tight junctions in Stria vascularis of the inner ear and to determine if the diversity of Claudin family members that comprise fight junctions located around Scala media have overlapping or non-overlapping properties. The knowledge gained from these studies will provide deep insights into the roles of the stria vascularis in hearing and potassium recycling to the endolymph. The specific aims are: 1) to test the hypothesis that inner ear pathology in Claudin//-null mice stems from morphological defects. Claudin 11 is expressed in the developing vestibulocochlear apparatus of embryos from E13.5 as well as in adults and evidence of gross structural changes during development and postnatally that could account for the pathology in the knockout mice will be sought. 2) to test the hypothesis that Claudin 11-null mice exhibit early hearing defects. Young adult knockout mice exhibit elevated auditory brainstem response (ABR) thresholds which indicate hearing loss, Further testing of ABRs as well as measurements of endocochlear potentials and DPOAEs at different ages will reveal if this phenotype is progressive or present at the time mice normally start to hear. 3) to optimize a transgene cassette for the expression of heterologous genes in basal cells of Stria vascularis. Basal cell-specific enhancer elements already in-hand will be located within the Claudin 11 gene and used to drive expression of a beta-galactosidase reporter gene in the cochlea, Reporter expression will be characterized with a view to expressing several claudin cDNAs in the cochlea in attempts to rescue hearing loss in Claudin 11-null mice.

描述（由申请人提供）：该项目的长期目标是定义内耳血管纹中紧密连接的功能，并确定包含位于 Scala 介质周围的战斗连接的 Claudin 家族成员的多样性是否具有重叠或不重叠的特性。从这些研究中获得的知识将深入了解血管纹在听力和钾再循环到内淋巴中的作用。具体目标是：1) 检验 Claudin//-null 小鼠内耳病理源于形态缺陷的假设。 Claudin 11 在 E13.5 胚胎发育中的前庭蜗器以及成人中表达，并且将寻找发育期间和出生后总体结构变化的证据，以解释敲除小鼠的病理学。 2）检验Claudin 11缺失小鼠表现出早期听力缺陷的假设。年轻的成年敲除小鼠表现出较高的听觉脑干反应（ABR）阈值，表明听力损失。进一步测试 ABR 以及测量不同年龄的耳蜗电位和 DPOAE 将揭示这种表型是进行性的还是在小鼠正常开始听力时出现。 3)优化用于在血管纹基底细胞中表达异源基因的转基因盒。现有的基底细胞特异性增强子元件将位于 Claudin 11 基因内，并用于驱动 β-半乳糖苷酶报告基因在耳蜗中的表达。报告基因的表达将被表征，以期在耳蜗中表达多个 Claudin cDNA，以试图挽救 Claudin 11 缺失小鼠的听力损失。