Modern Studies of the Human Temporal Bone

人类颞骨的现代研究

• 批准号: 7477061
• 负责人: Akira Ishiyama
• 金额: $ 39.16万
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-09-20 至 2011-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7477061
• 关键词: Acoustic Nerve; Antigens; Audiology; Auditory; Autopsy; Base Sequence; Biological; Biological Markers; Biological Preservation; Bone Diseases; Caliber; Cell Count; Cells; Classification; Clinical; Clinical assessments; Collagen; Connexins; Consent; Consult; Count; Coupled; DNA; Data; Databases; Development; Disease; Doctor of Medicine; Electron Microscopy; Epithelial; Equilibrium; Fluorescence; Formalin; Functional disorder; Future; Ganglia; Gene Expression; Genes; Genetic Transcription; Guidelines; Hair Cells; Hearing; Histopathology; Homeostasis; Human; Image; Imagery; Immunohistochemistry; In Situ Hybridization; Label; Laboratories; Laboratory Research; Labyrinth; Lasers; Ligaments; Light; Localized; Measures; Medical Records; Messenger RNA; Methodology; Methods; Microdissection; Modems; Molecular; Molecular Biology; Myxoid cyst; Nerve Fibers; Neuroanatomy; Neurology; Neurons; Neurosciences; Nucleic Acids; Nucleic acid sequencing; Organ; Otolaryngology; Paraffin Embedding; Pathogenesis; Pathology; Patients; Pattern; Principal Investigator; Process; Prospective Studies; Proteins; Protocols documentation; Questionnaires; RNA; Recording of previous events; Records; Recruitment Activity; Registries; Research; Research Personnel; Research Proposals; Resources; Science; Scientist; Sodium Channel; Specimen; Stria Vascularis; Structural Protein; Students; Supporting Cell; Susceptibility Gene; System; Techniques; Temporal bone structure; Testing; Tissues; Training; Training Programs; Update; Vestibular Diseases; Vestibular Function Tests; Vestibular Hair Cells; base; cell type; design; experience; improved; interest; laser capture microdissection; member; morphometry; neuroepithelium; novel; programs; prospective; protein expression; protocol development; sample fixation; spiral ganglion; tissue processing; water channel

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.

描述（由申请人提供）：人类颞骨研究对我们理解听力和平衡障碍的病理生理学做出了重大贡献。然而，尽管以前的努力，以阐明参与颞骨疾病的发病机制，有一个巨大的需求，以创建和研究颞骨标本，从患者明确的临床听觉和前庭疾病谁经历了定量的听觉和前庭功能测试，并由经验丰富的neurotologists检查。我们认为，颞骨科学的进步，以阐明听前庭疾病背后的分子和细胞病理生理学，需要一个系统的，前瞻性的临床病理学相关的方法，加上快速固定和组织处理协议，旨在抗原和形态保存，以及应用最现代化的准确和可靠的方法来定量和鉴定蛋白质和mRNA的病理变化基因表达。为了实现这一目标，我们在目标1中提出，通过招募捐赠者并获得详细的临床病史和对所有那些承诺颞骨的测试来扩大国家颞骨听力和平衡资源登记处，在目标2中提出，进一步开发新的人类颞骨处理技术，以最大限度地保留形态和抗原以及核酸，在目标3中提出，应用免疫组织化学的现代技术，无偏体视学，原位杂交和提取RNA的显微解剖的人颞骨标本。此外，还将进行传统的组织病理学相关性分析。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，我们计划在颞骨科学和神经耳科学方面培养临床科学家和博士后学生。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的RNA提取，并与艾伦瑞安博士合作在福尔马林固定石蜡包埋切片上应用原位杂交。