Modern Studies of the Human Temporal Bone

描述(申请人提供)：人类颞骨研究对我们理解听力和平衡障碍的病理生理学有很大贡献。然而，尽管以前曾努力阐明颞骨疾病的潜在机制，但仍有巨大的需要创建和研究来自明确临床听力和前庭疾病患者的颞骨标本，这些患者接受了定量的听觉和前庭功能测试，并由经验丰富的神经科医生进行检查。我们认为，要发展颞骨科学来阐明听前庭疾病背后的分子和细胞病理生理学，需要一种系统的、前瞻性的临床病理相关方法，结合以抗原性和形态保存为目的的快速固定和组织处理方案，以及应用最现代、准确和可靠的方法来量化和鉴定蛋白质和mRNA基因表达的病理变化。为了实现这一目标，我们在目标1中建议通过招募捐赠者和获得详细的临床病史并对所有承诺捐赠者的颞骨进行测试来扩大国家颞骨听力与平衡资源注册表，目标2进一步开发新的人类颞骨处理技术，以最大限度地保存形态、抗原和核保存，目标3应用免疫组织化学、无偏倚体视学、原位杂交和显微解剖人颞骨标本的RNA提取等现代技术。此外，还将进行传统的组织病理学相关研究。将分析DNA中的易感基因以及DMA和RNA表达之间的相关性。在目标4中，我们计划培训临床科学家和博士后学生有关颞骨科学和神经学的知识。在目标5中，我们计划系统地对图像进行分类和数字化，以实现数据共享，并计划与颞骨研究联盟充分合作，并与其他颞骨研究实验室合作。我们将与Kumar Alagramam博士合作开发使用激光捕获显微切割技术提取RNA，并与Allen Ryan博士合作在福尔马林固定的石蜡包埋切片上应用原位杂交。

DESCRIPTION (provided by applicant): Human temporal bone studies have contributed significantly to our understanding of the pathophysiology of hearing and balance disorders. However, despite previous efforts to elucidate the underlying mechanisms of involved in the pathogenesis of temporal bone disorders, there is a tremendous need to create and study temporal bone specimens from patients with well-defined clincial auditory and vestibular disorders who have undergone quantitative auditory and vestibular function testing, and examination by experienced neurotologists. We propose that the advancement of temporal bone science to elucidate the molecular and cellular pathophysiology behind audiovestibular diseases requires a systematic, prospective clinicopathological correlative approach, coupled with rapid fixation and tissue-processing protocols aimed at antigenic and morphological preservation, and the application of the most modern accurate and reliable methodology for quantification and identification of pathological changes in protein and mRNA gene expression. To accomplish this, we propose in aim 1 to expand the National Temporal Bone Hearing and Balance Resource Registry by recruiting donors and obtaining detailed clinical histories and testing on all those pledging temporal bones, in aim 2 to further develop novel human temporal bone processing techniques to maximize morphologic and antigenic and nucleic acie preservation, in aim 3 to apply the modern techniques of immunohistochemistry, unbiased stereology, in situ hybridization and extraction of RNA on microdissected human temporal bone specimens. Additionally, traditional histopathological correlations will be made. DNA will be analyzed for susceptibility genes and correlations between DMA and RNA expression. In aim 4, we plan to train clinician-scientists and postdoctoral students in temporal bone science and neurotology. In aim 5, we plan to systematically categorize and digitalize images to allow data sharing, and plan to cooperate fully with the Temporal Bone Research Consortium and to collaborate with other temporal bone research laboratories. We will collaborate with Dr. Kumar Alagramam in the development of RNA extraction using laser capture microdissection, and with Dr. Allen Ryan in the application of in situ hybridization on formalin-fixed paraffin-embedded sections.