Actin, gelsolin and filamin-A are dynamic actors in the cytoskeleton remodelling contributing to the blood brain barrier phenotype

The brain vascular endothelium operates as a dynamic regulatory interface to maintain the cell environment of the nervous system. In the vicinity of astrocytes, brain endothelial cells develop characteristic features conferring a strong cellular impermeability which limits the penetration of various...

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Bibliographic Details
Published in:Proteomics (Weinheim) 2009-03, Vol.9 (5), p.1207-1219
Main Authors: Pottiez, Gwënaël, Sevin, Emmanuel, Cecchelli, Roméo, Karamanos, Yannis, Flahaut, Christophe
Format: Article
Language:English
Subjects:
2-D PAGE
Actin-binding proteins
Actins - analysis
Analytical, structural and metabolic biochemistry
Animals
Astrocytes - cytology
Astrocytes - metabolism
Biochemistry, Molecular Biology
Biological and medical sciences
Blood-brain barrier
Blood-Brain Barrier - metabolism
Brain - blood supply
Brain - cytology
Cattle
Cell Differentiation
Cells, Cultured
Cerebral endothelial cells
Coculture Techniques - methods
Contractile Proteins - analysis
Cytoskeleton - metabolism
Endothelial Cells - cytology
Endothelial Cells - metabolism
Filamins
Fundamental and applied biological sciences. Psychology
Gelsolin - analysis
Life Sciences
Microfilament Proteins - analysis
Miscellaneous
Proteins
Proteome - analysis
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Summary:The brain vascular endothelium operates as a dynamic regulatory interface to maintain the cell environment of the nervous system. In the vicinity of astrocytes, brain endothelial cells develop characteristic features conferring a strong cellular impermeability which limits the penetration of various compounds. The aim of our study was to determine by differential proteomic analysis the changes occurring in bovine brain capillary endothelial cells (BBCEC) differentiated in co-culture with astrocytes compared with endothelial cells cultured alone. In order to obtain reproducible and meaningful protein profiles of in vitro blood-brain barrier models, three sample preparation procedures were carried out to provide the first 2-D comparative proteomic study of BBCEC. Our study highlights advantages and drawbacks of each procedure. The cellular proteins prepared from mechanical scraping of collagen-seeded BBCEC were strongly contaminated by serum proteins. Enzymatic dissociation of BBCEC by trypsin or collagenase solved this problem. A comparative 2-DE profile study of collagenase-harvested BBCEC revealed that cytoskeleton-related proteins (actin, gelsolin and filamin-A) show the most significant quantitative changes in the Triton soluble protein fraction from BBCEC that exhibit characteristics closest to the in vivo situation.
ISSN:1615-9853
1615-9861
DOI:10.1002/pmic.200800503