CORE--TISSUE CHARACTERIZATION AND IMMUNOREAGENT RESOURCE

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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.