title: A differential proteomic approach identifies structural and functional components that contribute to the differentiation of brain capillary endothelial cells.
abstract: When in the vicinity of astrocytes, brain capillary endothelial cells (BCECs) develop the characteristic structural and functional features of the blood-brain barrier (BBB). The latter has low cellular permeability and restricts various compounds from entering the brain. We recently reported that the cytoskeleton-related proteins actin, gelsolin and filamin-A undergo the largest quantitative changes in bovine BCECs after re-induction of BBB functions by co-culture with glial cells. In the present study, we used an in-depth, proteomic approach to quantitatively compare differences in Triton-X-100-solubilized proteins from bovine BCECs with limited or re-induced BBB functions (i.e. cultured in the absence or presence of glial cells, respectively). The 81 protein spots of differing abundance were linked to 55 distinct genes. According to the Protein ANalysis THrough Evolutionary Relationships classification system and an Ingenuity Pathway Analysis, these quantitative changes mainly affected proteins involved in (i) cell structure and motility and (ii) protein metabolism and modification. The fold-changes affecting HSPB1, moesin and ANXA5 protein levels were confirmed by western blot analysis but were not accompanied by changes in the corresponding mRNA expression levels. Our results reveal that the bovine BCECs' phenotype adaptation to variations in their environment involves the reorganization of the actin cytoskeleton.
authors: Univ Lille Nord de France, F-59000 Lille, France. gwenael.pottiez@gmail.com
authors: Gwënaël
authors: G
authors: Pottiez
authors: Sophie
authors: S
authors: Duban-Deweer
authors: Barbara
authors: B
authors: Deracinois
authors: Fabien
authors: F
authors: Gosselet
authors: Luc
authors: L
authors: Camoin
authors: Johan
authors: J
authors: Hachani
authors: Pierre-Olivier
authors: PO
authors: Couraud
authors: Roméo
authors: R
authors: Cecchelli
authors: Marie-Pierre
authors: MP
authors: Dehouck
authors: Laurence
authors: L
authors: Fenart
authors: Yannis
authors: Y
authors: Karamanos
authors: Christophe
authors: C
authors: Flahaut
mesh_terms: D000199:Actins / Q000235:genetics; D000818:Animals; D017302:Annexins / Q000235:genetics; D001812:Blood-Brain Barrier / Q000166:cytology / Q000502:physiology*; D001921:Brain / Q000098:blood supply*; D002417:Cattle; D002454:Cell Differentiation / Q000502:physiology*; D018920:Coculture Techniques; D015180:Electrophoresis, Gel, Two-Dimensional; D042783:Endothelial Cells / Q000166:cytology* / Q000502:physiology; D055551:HSP27 Heat-Shock Proteins / Q000235:genetics; D008840:Microfilament Proteins; D009457:Neuroglia / Q000166:cytology; D060066:Protein Interaction Maps; D040901:Proteomics / Q000379:methods; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D014746:Vimentin / Q000235:genetics
chemical_list: D000199:Actins; D017302:Annexins; D055551:HSP27 Heat-Shock Proteins; D008840:Microfilament Proteins; D012333:RNA, Messenger; D014746:Vimentin; C071058:moesin