CORE--ELECTRON MICROSCOPY

核心--电子显微镜

• 批准号: 6336300
• 负责人: WILLIAM A HORTON
• 金额: $ 18.82万
• 依托单位: UNIVERSITY OF TX MD ANDERSON CAN CTR
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-08-15 至 2001-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6336300
• 关键词: animal tissue; bioimaging /biomedical imaging; biomedical facility; electron microscopy; histology; immunoelectron microscopy; laboratory mouse; scanning transmission electron microscopy; skeletal system; transmission electron microscopy

Transgenic and gene targeting technologies will be used in this program project to generate mice strains in which the functions of several genes key to skeletal development and growth are disrupted. Ultrastructural analysis of skeletal tissues from these mice and their wild type littermates will be performed to help characterize these disturbances and delineate the normal functions of the gene products involved. The analyze will focus on the ultrastructure of epiphyseal cartilage, growth plate and bone cells and the structure and organization of matrices assembled in these locations. Established protocols will be employed. The electron microscopy (EM) core in Portland will carry out transmission EM and, when appropriate, scanning EM and immuno(gold) EM studies on the murine skeletal tissues and rotary shadowing of fibrillar constituents of the skeletal matrices. In some instances, i.e., transmission EM, mouse skeletal tissues will be fixed in Houston and shipped to Portland. In other, i.e., immuno EM, mice will be transferred to Portland so that the tissues of interest can be exposed to antibody before fixation. All of these protocols are currently in operation in the facility, and a wide range of antibodies that recognize murine skeletal protein epitopes are available. The investigators, Dr. William Horton and Doug Keene, have a long standing interest and special expertise in this field, and the EM facility in Portland is particularly suited for the proposed studies. Moreover, both investigators have collaborated closely and productively with other PIs in the program project demonstrating how mice, mouse embryos, murine skeletal tissues and information can be easily transferred from one location to the other as well as the effectiveness of this working arrangement. The ultrastructural analyzes to be provided by this core are essential to the overall performance on the program project.

转基因和基因靶向技术将用于本项目中，以产生几种骨骼发育和生长关键基因功能被破坏的小鼠品系。将对这些小鼠及其野生型幼崽的骨骼组织进行超微结构分析，以帮助表征这些紊乱并描绘相关基因产物的正常功能。分析将集中在骨骺软骨、生长板和骨细胞的超微结构以及在这些位置组装的基质的结构和组织。将采用既定的协议。波特兰的电子显微镜（EM）核心将对小鼠骨骼组织进行透射电子显微镜研究，并在适当的时候进行扫描电子显微镜和免疫（金）电子显微镜研究，并对骨骼基质的纤维成分进行旋转阴影。在某些情况下，例如，传播EM，小鼠骨骼组织将在休斯敦固定并运往波特兰。在另一种情况下，即免疫电镜，小鼠将被转移到波特兰，使感兴趣的组织可以在固定前暴露于抗体。所有这些方案目前都在该设施中运行，并且有广泛的识别小鼠骨骼蛋白表位的抗体可用。研究人员William Horton博士和Doug Keene博士在这一领域有着长期的兴趣和特殊的专业知识，波特兰的EM设施特别适合拟议的研究。此外，两位研究人员在项目项目中与其他pi密切合作，展示了小鼠，小鼠胚胎，小鼠骨骼组织和信息如何轻松地从一个位置转移到另一个位置，以及这种工作安排的有效性。该核心提供的超微结构分析对项目的整体性能至关重要。