This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Two proteins of great interest in the research on the degenerative autoimmune disease Multiple Sclerosis are Myelin OligodendroGlial (MOG) and Proteo Lipid Protein (PLP). Transport of raft-like structures may be involved in maintaining the myelin layers. OLN-93 oligodendroglial cells in all stages of development are used to image with one-photon confocal, FCS, FRAP, FRET and TIRF microscopy the transiently transfected MOG-eGFP and PLP-eGFP constructs and currently also stably transfected MOG-eGFP constructs. MOG is located in the uncompacted outer perimeter of the cells and in the processes (extensions). PLP forms part of the compacted myelin. Since the cells move about single point FCS analysis analysis is hampered by this motion. Current visit plans to explore and get training in the application of the state-of-the-art and very promising RICS analysis of already collected timeseries and to obtain advice how to optimize image collection procedures. To this end also images collected on MDCK and CHO cells related to current research on cell stress (Metabolic Inhibition) effects on cytosolic and mitochondrial ion concentrations, pH, and mitochondrial potential are being analyzed for comparison with results obtained with electrophysiological methods. This research is of prime importance to and is part of the strategy at Hasselt University to develop within a European framework biosensors based on new materials like functionalized plastics and diamond surfaces.
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