CORE--ELECTRON MICROSCOPY

核心--电子显微镜

• 批准号: 6443314
• 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: 2001
• 资助国家: 美国
• 起止时间: 2001-04-01 至 2002-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6443314
• 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.

该项目将使用转基因和基因打靶技术来培育小鼠品系，其中对骨骼发育和生长至关重要的几个基因的功能被破坏。对这些小鼠及其野生型同窝小鼠的骨骼组织进行超微结构分析，以帮助表征这些干扰并描绘所涉及基因产物的正常功能。该分析将重点关注骨骺软骨、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。将采用既定协议。波特兰的电子显微镜 (EM) 核心将进行透射电镜，并在适当时对小鼠骨骼组织进行扫描电镜和免疫（金）电镜研究，并对骨骼基质的纤维成分进行旋转阴影研究。在某些情况下，即传输 EM，小鼠骨骼组织将在休斯顿固定并运往波特兰。在其他情况下，即免疫 EM，小鼠将被转移到波特兰，以便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，并且有多种识别小鼠骨骼蛋白表位的抗体可供使用。研究人员 William Horton 博士和 Doug Keene 博士对该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，两位研究人员与该项目中的其他 PI 进行了密切而富有成效的合作，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于程序项目的整体性能至关重要。