title: TNAP and EHD1 are over-expressed in bovine brain capillary endothelial cells after the re-induction of blood-brain barrier properties.
abstract: Although the physiological properties of the blood-brain barrier (BBB) are relatively well known, the phenotype of the component brain capillary endothelial cells (BCECs) has yet to be described in detail. Likewise, the molecular mechanisms that govern the establishment and maintenance of the BBB are largely unknown. Proteomics can be used to assess quantitative changes in protein levels and identify proteins involved in the molecular pathways responsible for cellular differentiation. Using the well-established in vitro BBB model developed in our laboratory, we performed a differential nano-LC MALDI-TOF/TOF-MS study of Triton X-100-soluble protein species from bovine BCECs displaying either limited BBB functions or BBB functions re-induced by glial cells. Due to the heterogeneity of the crude extract, we increased identification yields by applying a repeatable, reproducible fractionation process based on the proteins' relative hydrophobicity. We present proteomic and biochemical evidence to show that tissue non-specific alkaline phosphatase (TNAP) and Eps15 homology domain-containing protein 1(EDH1) are over-expressed by bovine BCECs after the re-induction of BBB properties. We discuss the impact of these findings on current knowledge of endothelial and BBB permeability.
authors.affiliation: Université Lille Nord de France, France.
authors.forename: Barbara
authors.initials: B
authors.lastname: Deracinois
authors.forename: Sophie
authors.initials: S
authors.lastname: Duban-Deweer
authors.forename: Gwënaël
authors.initials: G
authors.lastname: Pottiez
authors.forename: Roméo
authors.initials: R
authors.lastname: Cecchelli
authors.forename: Yannis
authors.initials: Y
authors.lastname: Karamanos
authors.forename: Christophe
authors.initials: C
authors.lastname: Flahaut
mesh_terms: D000469:Alkaline Phosphatase / Q000235:genetics* / Q000378:metabolism; D000818:Animals; D001812:Blood-Brain Barrier / Q000737:chemistry / Q000378:metabolism*; D001921:Brain / Q000378:metabolism*; D002417:Cattle; D002478:Cells, Cultured; D042783:Endothelial Cells / Q000737:chemistry / Q000378:metabolism*; D004789:Enzyme Activation / Q000187:drug effects; D015870:Gene Expression; D007978:Levamisole / Q000494:pharmacology; D009457:Neuroglia / Q000378:metabolism; D040901:Proteomics; D051381:Rats; D033921:Vesicular Transport Proteins / Q000235:genetics* / Q000378:metabolism
chemical_list: C120700:EHD1 protein, human; D033921:Vesicular Transport Proteins; D007978:Levamisole; D000469:Alkaline Phosphatase