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

Gwënaël Pottiez 1 Emmanuel Sevin 1 Roméo Cecchelli 1 Yannis Karamanos 2 Christophe Flahaut 1
1 LBHE - Laboratoire de Physiopathologie de la Barrière Hémato-Encéphalique
2 Laboratoire de Biochimie Moléculaire et Cellulaire
Abstract : 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 coculture 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 cytoskeletonrelated 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.
Keywords : 2-D PAGE Actin-binding proteins Blood-brain barrier Cerebral endothelial cells
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Journal articles
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Submitted on : Monday, December 6, 2010 - 3:31:50 PM
Last modification on : Monday, December 6, 2010 - 3:31:50 PM

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Gwënaël Pottiez, Emmanuel Sevin, Roméo Cecchelli, Yannis Karamanos, Christophe Flahaut. Actin, gelsolin and filamin-A are dynamic actors in the cytoskeleton remodelling contributing to the blood brain barrier phenotype. Proteomics, Wiley-VCH Verlag, 2009, 9, p. 1207-1219. ⟨hal-00543716⟩

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