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Title: | Diffusion of sphingomyelin and myelin oligodendrocyte glycoprotein in the membrane of OLN-93 oligodendroglial cells studied by fluorescence correlation spectroscopy | Authors: | GIELEN, Ellen Vercammen, J Sykora, J Humpolickova, J VAN DE VEN, Martin Benda, A. HELLINGS, Niels Hof, M Engelborghs, Y STEELS, Paul AMELOOT, Marcel |
Issue Date: | 2005 | Publisher: | ELSEVIER FRANCE-EDITIONS SCIENTIFIQUES MEDICALES ELSEVIER | Source: | COMPTES RENDUS BIOLOGIES, 328(12). p. 1057-1064 | Abstract: | Evidence has been accumulated that the plasma membrane of various mammalian cell types is heterogeneous in structure and may contain lipid microdomains (lipid rafts). This study focuses on the membrane organization of living oligodendrocytes, which are the myelin-producing cells of the central nervous system. Fluorescence correlation spectroscopy (FCS) was used to monitor the lateral diffusion of a lipid and of a protein in the oligodendroglial cell line OLN-93. The lipid was fluorescently labelled sphingomyelin (Bodipy FL-C5 SM). The protein was the myelin oligodendrocyte glycoprotein (MOG). In order to monitor the lateral diffusion of MOG, OLN-93 cells were transfected with a MOG-EGFP (enhanced green fluorescent protein) fusion plasmid. The measurements were performed at room temperature. FCS data were analyzed for two-dimensional (2D) diffusion according to three models which all included a triplet fraction: (a) 2D I component (2D1C), (b) 2D anomalous diffusion (2D1C alpha), and (c) 2D 2 components (2D2C). Preliminary results indicate that for the lipid case, the best fits are obtained with 2D2C. In the case of MOG-EGFP, 2D2C and 2D1C alpha give fits of similar quality. The parameter estimates obtained with 2D1C alpha, however, have a lower standard deviation. The anomaly parameter for MOG-EGFP is 0.59 +/- 0.01. | Notes: | Hasselt Univ, Biomed Res Inst, tUL, B-3590 Diepenbeek, Belgium. Katholieke Univ Leuven, Dept Biochem, B-3001 Heverlee, Belgium. Acad Sci Czech Republ, J Heyrovsky Inst Phys Chem, CR-18223 Prague, Czech Republic.Ameloot, M, Hasselt Univ, Biomed Res Inst, tUL, Univ Campus Bldg D&A, B-3590 Diepenbeek, Belgium.marcel.ameloot@uhasselt.be | Keywords: | OLN-93; FCS; MOG; sphingomyelin; rafts; oligodendrocyte; anomalous diffusion | Document URI: | http://hdl.handle.net/1942/2108 | ISSN: | 1631-0691 | e-ISSN: | 1768-3238 | DOI: | 10.1016/j.crvi.2005.09.008 | ISI #: | 000233831300004 | Category: | A1 | Type: | Journal Contribution | Validations: | ecoom 2006 |
Appears in Collections: | Research publications |
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