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Preliminary Study on the Role of Lipid Raft’s Peculiar Proteins in Vesicular Transportation

Author: CaoHuiYa
Tutor: XuTao
School: Huazhong University of Science and Technology
Course: Biophysics
Keywords: Lipid rafts Nest Caveolin-1 Flotillin-1 TOPO vector Total internal reflection fluorescence microscopy imaging technology
CLC: Q25
Type: Master's thesis
Year: 2005
Downloads: 216
Quote: 0
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Abstract


The cell material transport is essentially a cell within a large and complex logistics system, the exception of this system will lead to the occurrence of the corresponding disease. Vesicles important means of transport is a regulation of the logistics system, lipid rafts is a micro-area containing a special lipid and protein within the membrane lipid bilayer, they have an important role in the logistics system. The topics introduced single-molecule research techniques and total internal reflection fluorescence imaging, real-time dynamic studies of dynamic spatial orientation relationship between the two types of lipid rafts protein and vesicle, and analysis of the dynamic characteristics of a lipid raft protein. This study helps us to further reveal the mechanism of lipid rafts; and has important theoretical and practical significance for the future, we further understanding of lipid rafts. This paper first constructed expression plasmid containing fluorescently labeled caveolin and flotillin two types of lipid rafts specific protein, a preliminary study of the relationship between lipid rafts and vesicle trafficking. This experiment, we specifically using total internal reflection fluorescence microscopy imaging techniques studied in the mouse adrenal pheochromocytoma cell line (PC12) of caveolin and NPY in the SNAP25 the intracellular dynamic positioning relationship and interaction as well as flotillin and NPY SNAP25 the the intracellular dynamic positioning relations and interactions. The results show that: the expression of caveolin whether at rest or in high potassium stimulation with the NPY large vesicles or with SNAP25 coexistence. Same flotillin the NPY large vesicles and SNAP25 coexistence and flotillin itself was a patchy distribution under TIRFM flotillin expression soon distributed in the cell membrane. Positioning relations from the point of view, the lipid raft marker protein no obvious evidence involved in vesicle trafficking. This is not the same as previously chemically extracted PC12 cells and found that constitute the SNARE protein SNAP-25 in the cholesterol-rich lipid rafts conclusion. Different reasons: on the one hand, may be caused by different experimental methods, on the other hand may be because the large vesicles transport does not require lipid rafts mediated. In addition, PC12 cells transfected caveolin-1-EGFP, using total internal reflection fluorescence microscopy imaging techniques vesicle movement records to carry caveolin-1 in 60mM K ~ before and after stimulation with a significant difference. At the same time, we also analyze the dynamic characteristics of the the caveolin mark a vesicle.

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CLC: > Biological Sciences > Cell Biology > Cell physiology
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