Integrated Project funded by the
European Community,
Framework Programme 6

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Core Technologies

1. Protein chemistry / Cell biology: Proteins encoded by disease genes can be characterized in terms of structure and function by state-of-the-art techniques in protein chemistry/cell biology. Proteins in question will be produced in expression systems, and probed for interactions with established interaction partners using fluorescence depolarization and BIAcore analysis. By yeast two-hybrid screening and affinity chromatography accompanied by mass spectormetry, novel molecular partakers will be identified and correlations between loss of function in animal models and molecular interactions in vitro will be shown. Proteins will be introduced into cell lines or organ cultures and tested for molecular interactions using biochemical techniques, confocal microscopy and fluorescence renosance energy transfer (FRET). Most partners have access to local state-of-the-art technologies in cell biology and protein chemistry.


2. Renal (Patho)physiology: Detailed phenotypic analysis of animal models with renal diseases will be essential to uncover new pathophysiological mechanisms of dissease processes. We have at our disposal sophisticated methods for characterization of renal (dys)function in many laboratory animal models (e.g., blood pressure, hemodynamics, glomerular and renal vascular functio, magnetic resonance imaging). In addition, morphological and pathophysiological analysis willl be performed on specimens including light-, confocal- and electron microscopy, in situ hybridization, immunohistochemistry, as well as electrophysiological and perfusion studies in cell and organ cultures. A large repertoire of morphological tchniques will be provided by Erik i. Christensen. Jürg Biber and Friedrich Luft will provide technologies for in depth renal phenotyping of rat and mouse mouse models.


3. Proteome / Transcriptome Analysis: Knowledge of qualitative and quantitative gene expression profiles is central to understanding the role of gene products in renal development and disease. We will apply novel expression profileing techniques to systematically study the expression of genes during developmental processes and in a state of disease. These techniques include automated high-throughput in situ hybridization on whole mount embryos and kidney sections, microarray analysis and two-dimensional protein gel electopphoresis combine d with mass spectrometry. Transciptome activities will be located in four dedicated centers (Max Delbrueck Center in Berlin, Max Planck Institute in Hannover, ETH Zurich, and MRC Human Genetics Unit in Edinburgh). Stephan Ohlmeier (University of Oulu) is fully dedicated to proteomics and will carry oyt a significant part of this work.


4. Genotyping / Mutation Detection: Several of the partners have efficient, relatively high throughput genotyping/mutation detection facilities (e.g., Gene Mapping Center at the Max Delbrueck Center). These will be made available to the other partners who wish to map loci or to screen available models fo mutations in candidate genes.


5. Diagnosis and Therapy: Our ultimate goal will be a better understanding of genetic mechanisms controlling renal development and disease, leading to improved methods of diagnosis and therapy. In collaboration with SME partners we will develop new diagnostic/research tools essential to research and patient care. These products include reagents (cell lines, models), diagnostic tools (antibodies, custom microarrays, transcriptome/proteome maps) and small molecule compounds for pharmacological intervention (agonists, antagonists). Drug screening services will be provided by RecetIcon ApS, Aarhus. Diagnostic tools will be developed through sub-contracting at WITA Proteomics, Berlin and AROS Applied Biotechnology, Aarhus.