Abstract
A global proteomics strategy was initiated to decipher molecular mechanisms associated with the blood-brain barrier (BBB) phenotype of the brain capillary endothelial cells. The different methods implemented were shown complementarily. The main results obtained using an in vitro BBB model allowed highlighting the role of several protein actors of cytoskeleton remodelling, the involvement of the asymmetric dimethylarginine pathway in regulating endothelial function and the role of tissue non-specific alkaline phosphatase in the regulation of endothelial permeability.
Highlights
The blood-brain barrier (BBB) corresponding to the cerebral capillaries covers approximately 95% of the total area of barriers between blood and brain [1]
The morphology and functional properties of the brain capillary endothelial cells (BCEC) that form with other cells (Figure 1) the BBB are well documented: a decrease in endothelial permeability, fewer caveolae, the reinforcement of tight junctions, fewer pinocytic vesicles, an increase in the number of mitochondria and a higher transendothelial electrical resistance [2,3]
Our results revealed that the BCEC can adapt to variations in their environment and this involves the reorganization of the actin cytoskeleton contributing to the blood-brain barrier phenotype
Summary
The blood-brain barrier (BBB) corresponding to the cerebral capillaries covers approximately 95% of the total area of barriers between blood and brain [1]. The tissue complexity cannot be reproduced with in vitro models, but in turn those models are more flexible, the medium can be renewed and complemented and large quantities of well characterised cells can be obtained The latter characteristic has formed the basis of the proteomics strategy described . The in vitro BBB model developed in our laboratory utilizes bovine BCEC that were purified to homogeneity by mechanical homogenisation and filtration [6] and subsequent co-culturing with murine glial cells. These particular culture conditions induce differentiation of endothelial cells which provide an in vivo BBB phenotype [7]. In the effort to decipher molecular mechanisms for the establishment of the BBB, we have experimented with several methods and we demonstrated that the in-gel [9,10] and the off-gel [9,11,12] approaches were complementary
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