CORE--TISSUE CHARACTERIZATION AND IMMUNOREAGENT RESOURCE

核心——组织表征和免疫试剂资源

• 批准号: 7069764
• 负责人: JOANNE E MURPHY-ULLRICH
• 金额: $ 16.05万
• 依托单位: UNIVERSITY OF ALABAMA AT BIRMINGHAM
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-09-30 至 2010-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7069764
• 关键词: animal tissue; autosomal recessive trait; biomedical facility; fluorescence resonance energy transfer; histochemistry /cytochemistry; immunocytochemistry; immunologic substance development /preparation; in situ hybridization; laser capture microdissection; light microscopy; molecular /cellular imaging; monoclonal antibody; morphometry; polycystic kidney; polymerase chain reaction; tissue /cell preparation; transmission electron microscopy

Understanding the molecular mechanisms underlying mouse models of autosomal recessive PKD (ARPKD) will require expert morphological and immunohistochemical analysis of renal, biliary, and hepatic tissues, access to state of the art molecular analyses for assessment of ARPKD cells, and generation of novel immunoreagents to probe ARPKD cell regulation. The goal of this Core is to provide a comprehensive array of services for molecular analysis of ARPKD tissues and cells through the coordination of three well-established UAB Core facilities in a single unified platform in a cost-effective manner to maximize accessibility, to optimize protocols, and to provide training for the local and extended ARPKD research base. The Tissue Characterization and Immunoreagent Resource (TCIR) Core will provide a centralized and controlled facility for processing of mouse tissues for morphologic and histochemical analysis, optimization of immunohistochemical protocols, and quantitative morphometric analysis. In addition, P30 investigators will have access to advanced technologies such as Laser Capture Microdissection (LCM), fluorescence resonance energy transfer (FRET) microscopy, and in situ hybridization. The Epitope Recognition and Immunoreagent Component (ERIC) of the TCIR Core will assist P30 investigators in the development of novel monoclonal reagents (hybridoma and antibody phage display technology) with relevance to the study of PKD and also serve as a production and distribution center for commonly utilized monoclonal antibodies. The overall objectives of the TCIR are: SPECIFIC AIM 1 - To provide accurate, reproducible, and standardized expert light microscopic pathologic interpretation of animal tissues provided by investigators. SPECIFIC AIM 2 - To provide for the processing and data accumulation to P30 investigators of animal tissues requiring sophisticated special techniques including morphometry, quantitative PCR, in-situ hybridization, immunohistochemistry, transmission electron microscopy, laser capture microdissection, and FRET live cell imaging. SPECIFIC AIM 3 -Provide PKD investigators with expertise in hybridoma and antibody phage display technology to facilitate production and distribution of novel or established monoclonal reagents related to the study of ARPKD. SPECIFIC AIM 4 -To enhance cost effectiveness by centralized purchasing and inventory management of consumable supplies and to efficiently perform quality control and quality assurance activities required by the local and extended research base.

了解常染色体隐性遗传性 PKD 小鼠模型的分子机制 (ARPKD) 将需要对肾、胆道和肝组织进行专家形态学和免疫组织化学分析，获得最先进的分子分析来评估 ARPKD 细胞，并生成新型免疫试剂来探测 ARPKD 细胞调节。该核心的目标是通过在一个统一平台上协调三个完善的 UAB 核心设施，以经济高效的方式最大限度地提高可访问性，优化方案，为 ARPKD 组织和细胞的分子分析提供全面的服务，并为当地和扩展的 ARPKD 研究基地提供培训。组织表征和免疫试剂资源 (TCIR) 核心将为小鼠组织处理提供集中和受控的设施，用于形态学和组织化学分析、免疫组织化学方案优化和定量形态分析。此外，P30 调查人员将能够使用激光捕获等先进技术 显微切割 (LCM)、荧光共振能量转移 (FRET) 显微镜和原位 杂交。 TCIR 核心的表位识别和免疫试剂组件 (ERIC) 将协助 P30 研究人员开发与 PKD 研究相关的新型单克隆试剂（杂交瘤和抗体噬菌体展示技术），并作为常见试剂的生产和分销中心。使用单克隆抗体。 TCIR 的总体目标是： 具体目标 1 - 为研究人员提供的动物组织提供准确、可重复和标准化的专家光学显微镜病理解释。具体目标 2 - 为 P30 研究人员提供需要复杂特殊技术的动物组织处理和数据积累，包括形态测量、定量 PCR、原位杂交、免疫组织化学、透射电子显微镜、激光捕获显微切割和 FRET 活细胞成像。具体目标 3 - 为 PKD 研究人员提供杂交瘤和抗体噬菌体展示技术方面的专业知识，以促进与 ARPKD 研究相关的新型或已建立的单克隆试剂的生产和分销。 具体目标 4 - 通过消耗品的集中采购和库存管理来提高成本效益，并有效地执行本地和扩展研究基地所需的质量控制和质量保证活动。