Modern Studies of the Human Temporal Bone

描述（由申请人提供）：人类颞骨的研究对我们对听力和平衡障碍的病理生理学的理解做出了重大贡献。然而，尽管之前已经努力阐明了颞骨疾病发病机制的潜在机制，但仍然非常需要从明确的临床听觉和前庭功能障碍患者中创建和研究颞骨标本，这些患者已经接受了定量的听觉和前庭功能测试，并由经验丰富的神经学家进行检查。我们建议，颞骨科学的进步，以阐明听骨前庭疾病背后的分子和细胞病理生理，需要一个系统的、前瞻性的临床病理相关方法，加上快速固定和组织处理方案，旨在抗原和形态保存。以及应用最现代、最准确、最可靠的方法来定量和鉴定蛋白质和mRNA基因表达的病理变化。为了实现这一目标，我们建议在目标1中通过招募捐赠者，获取详细的临床病史和对所有承诺的颞骨进行测试来扩大国家颞骨听力和平衡资源注册，在目标2中进一步开发新的人类颞骨处理技术，以最大限度地提高形态学，抗原和核酸保存，在目标3中应用免疫组织化学，无偏体视学的现代技术，显微解剖人类颞骨标本的原位杂交和RNA提取。此外，将进行传统的组织病理学关联。将分析DNA的易感基因以及DMA与RNA表达之间的相关性。在目标4中，我们计划培养颞骨科学和神经学方面的临床科学家和博士后。在目标5中，我们计划系统地对图像进行分类和数字化，以允许数据共享，并计划与颞骨研究联盟和其他颞骨研究实验室充分合作。我们将与Kumar Alagramam博士合作开发利用激光捕获显微解剖提取RNA的技术，并与Allen Ryan博士合作在福尔马林固定石蜡包埋切片上应用原位杂交技术。

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.