PROTEINS OF ORGAN OF CORTI AND SURROUNDING FLUIDS

柯蒂氏器官和周围液体的蛋白质

• 批准号: 3218095
• 负责人: RUEDIGER R THALMANN
• 金额: $ 32.34万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1992
• 资助国家: 美国
• 起止时间: 1992-09-01 至 1995-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3218095
• 关键词: SDS polyacrylamide gel electrophoresis; cerebrospinal fluid; developmental neurobiology; endolymph; enzyme linked immunosorbent assay; gene expression; immunoaffinity chromatography; immunocytochemistry; molecular cloning; organ culture; organ of Corti; perilymph; polymerase chain reaction; protein purification; protein sequence; protein structure function

The ear converts acoustic impulses into signals interpretable by higher centers. The delicate acoustic instrument, the organ of Corti (0C) is made of living cells with all their susceptibilities. Most dysfunctions of the ear are mediated chemically, and for medical reasons, it is Imperative that these biochemical mysteries be elucidated. This proposal considers the biochemistry and molecular biology of two proteins (OCPI and OCP2) of unknown function present in the 0C at exceedingly high concentrations. The large amounts, expression restricted to the inner ear, and major increase in expression at the time the cochlea begins to function imply an important role in audition. This dictates an aggressive interdisciplinary approach to elucidate their function. We will apply standard techniques of protein chemistry adapted to the micro level. Analysis of primary amino acid sequence will exploit recent technological advances. Cellular and subcellular localization by immunohistochemical techniques will provide indirect information about their function. Standard cloning techniques will provide fundamental insights into structure and function of the OCPs. More importantly, the proposed mRNA expression experiments use OCPs as a tool to obtain important information about inner ear development. Cloning human 0CP genes provides them as candidate genes for inner ear disease. A large body of preliminary information on these projects makes them high priority and low risk. The 0C resides in tissue culture, as it were, surrounded by fluids of fundamentally different chemical makeup and electrical polarization. A precise definition of the composition of these fluids is of paramount importance. We have designed sophisticated techniques to avoid artifacts in the determination of the chemical composition of perilymph. We will determine valid concentrations of numerous small molecules and establish comprehensive protein profiles. Most artifacts are due to influx of cerebrospinal fluid (CSF) whenever the otic capsule is perforated, either for the sampling of specimens or insertion of electrodes. The influx of CSF may damage the 0C, since it results in very low levels of amino acids, low protein, and other potential sequelae. This situation also exists in perilymph fistulae, and may contribute to the disturbance of hearing. Our studies are also geared to the study of perilymph- or disease-specific proteins, and will critically evaluate the claim that perilymph of scala tympani is derived in bulk from CSF.

耳朵将声音脉冲转换成信号， 中心. 精巧的声学乐器，科尔蒂管风琴（0C）， 由具有所有生物学特性的活细胞组成。 大多数功能障碍 的耳朵是介导的化学，并为医学原因，它是 我们迫切需要解开这些生化之谜。 这项建议 考虑了两种蛋白质（OCPI）的生物化学和分子生物学 和OCP 2）的未知功能，存在于OC中，在极高的 浓度的 大量的，表达局限于内部 耳朵，并且在耳蜗开始时表达大幅增加 功能意味着在听觉中的重要作用。 这就要求 积极的跨学科方法来阐明其功能。 我们 将应用蛋白质化学的标准技术， 水平 一级氨基酸序列的分析将利用最近的 技术进步。 细胞和亚细胞定位， 免疫组织化学技术将提供间接信息， 它们的功能。 标准的克隆技术将提供基本的 OCP的结构和功能。 更重要的是 提出的mRNA表达实验使用OCP作为工具， 关于内耳发育的重要信息。 克隆人0CP 基因提供了它们作为内耳疾病的候选基因。 大 关于这些项目的初步信息使他们很高 优先和低风险。 可以说，0C存在于组织培养中， 周围都是化学成分完全不同的液体 电极化 一个精确的定义的组成 这些流体是至关重要的。 我们设计了复杂的 技术，以避免在确定的化学品 外淋巴液的组成。 我们将确定有效浓度 许多小分子和建立全面的蛋白质谱。 大多数伪影是由于脑脊液（CSF）的流入， 耳囊穿孔，用于样本取样或 插入电极。 CSF的流入可能会损害OC，因为它 导致非常低的氨基酸水平，低蛋白质，和其他 潜在的后遗症 这种情况也存在于外淋巴瘘， 并可能导致听力障碍。 我们的研究也 适用于外淋巴或疾病特异性蛋白质的研究，并将 批判性地评价声阶外淋巴来源于 从脑脊液中大量提取