DIFFUSION OF LIPIDS AND PROTEINS IN THE MEMBRANES OF OLIGODENDROGLIAL CELLS

该子项目是利用NIH/NCRR资助的中心赠款提供的资源的许多研究子项目之一。子项目和研究者（PI）可能从另一个NIH来源获得主要资金，因此可以在其他CRISP条目中表示。所列机构为中心机构，不一定为研究者机构。在退行性自身免疫性疾病多发性硬化的研究中，两种非常感兴趣的蛋白质是髓鞘少突胶质细胞（MOG）和蛋白脂质蛋白（PLP）。筏状结构的运输可能参与维持髓鞘层。将所有发育阶段的OLN-93少突胶质细胞用于用单光子共聚焦、FCS、FRAP、FRET和TIRF显微镜对瞬时转染的MOG-eGFP和PLP-eGFP构建体以及目前还稳定转染的MOG-eGFP构建体成像。MOG位于细胞未压实的外周和突起（延伸部分）中。PLP构成致密髓鞘的一部分。由于细胞在单点上移动，FCS分析受到这种运动的阻碍。目前的访问计划探索和获得培训，应用最先进的和非常有前途的RICS分析已经收集的时间序列，并获得如何优化图像收集程序的建议。为此，还分析了在MDCK和CHO细胞上收集的与细胞应激（代谢抑制）对细胞溶质和线粒体离子浓度、pH值和线粒体电位影响的当前研究相关的图像，以与电生理学方法获得的结果进行比较。 这项研究对于哈瑟尔特大学在欧洲框架内开发基于功能化塑料和金刚石表面等新材料的生物传感器战略至关重要，也是该战略的一部分。

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.