“…Therefore it is likely that modulation of its synthesis, and localization at the plasma membrane is of crucial importance in the establishment and maintenance of a functional BBB [22][23][24][25][26][27].…”
Section: Discussionmentioning
confidence: 99%
“…In these conditions, RBE4.B form a barrier with BBB permeability properties [22]. As also astrocytes are known to induce and maintain BBB properties in endothelial cells [23][24][25], we set a three-cell type-culture system that includes RBE4.B brain capillary endothelial cells, astrocytes and neurons.…”
Brain capillary endothelial cells form a functional barrier between blood and brain, based on the existence of tight junctions that limit paracellular permeability. Occludin is one of the major transmembrane proteins of tight junctions and its peripheral localization gives indication of tight junction formation. We previously reported that RBE4.B cells (brain capillary endothelial cells), cultured on collagen IV, synthesize occludin and correctly localize it at the cell periphery only when cocultured with neurons. In the present study, we describe a three‐cell type‐culture system that allowed us to analyze the combined effects of neurons and astrocytes on differentiation of brain capillary endothelial cells in culture. In particular, we found that, in the presence of astrocytes, the neuron‐induced synthesis and localization of occludin is precocious as compared to cells cocultured with neurons only.
“…Therefore it is likely that modulation of its synthesis, and localization at the plasma membrane is of crucial importance in the establishment and maintenance of a functional BBB [22][23][24][25][26][27].…”
Section: Discussionmentioning
confidence: 99%
“…In these conditions, RBE4.B form a barrier with BBB permeability properties [22]. As also astrocytes are known to induce and maintain BBB properties in endothelial cells [23][24][25], we set a three-cell type-culture system that includes RBE4.B brain capillary endothelial cells, astrocytes and neurons.…”
Brain capillary endothelial cells form a functional barrier between blood and brain, based on the existence of tight junctions that limit paracellular permeability. Occludin is one of the major transmembrane proteins of tight junctions and its peripheral localization gives indication of tight junction formation. We previously reported that RBE4.B cells (brain capillary endothelial cells), cultured on collagen IV, synthesize occludin and correctly localize it at the cell periphery only when cocultured with neurons. In the present study, we describe a three‐cell type‐culture system that allowed us to analyze the combined effects of neurons and astrocytes on differentiation of brain capillary endothelial cells in culture. In particular, we found that, in the presence of astrocytes, the neuron‐induced synthesis and localization of occludin is precocious as compared to cells cocultured with neurons only.
Blood-brain barrier (BBB) is a selective and dynamic structure, formed by endothelial cells, that allows the selective passage of substances from blood to CNS and vice versa. During brain development, brain capillary endothelial cells (BCECs) gradually acquire the ability to form a selective barrier, and it is known that astrocytes
AbstractWe previously found that RBE4.B brain capillary endothelial cells (BCECs) form a layer with blood-brain barrier (BBB) properties if co-cultured with neurons for at least one week. As astrocytes are known to modulate BBB functions, we further set a culture system that included RBE4.B BCECs, neurons and astrocytes. In order to test formation of BBB, we measured the amount of 3 H-sucrose able to cross the BCEC layer in this three-cell type model of BBB. Herein we report that both neurons and astrocytes induce a decrease in the permeability of the BCEC layer to sucrose. These effects are synergic as if BCECs are cultured with both neurons and astrocytes for 5 days, permeability to sucrose decreases even more. By Western analysis, we also found that, in addition to the canonical 60 kDa occludin, anti-occludin antibodies recognize a smaller protein of 48 kDa which accumulates during rat brain development. Interestingly this latter protein is present at higher amounts in endothelial cells cultured in the presence of both astrocytes and neurons, that is in those conditions in which sucrose permeation studies indicate formation of BBB.
“…We previously found that both neurons (24,27,28) and astrocytes (23,24) influence the ability of endothelial cells to form a barrier with properties resembling those of BBB. In the present study, we used an already described BBB in vitro model (23,24) to investigate whether the serum from patients with secondary progressive multiple sclerosis (SPMS) (29) caused any alteration of the neuronal vitality and morphology, and/or any BBB damage.…”
Abstract. An important component of the pathogenic process of multiple sclerosis (MS) is the blood-brain barrier (BBB) damage. We recently set an in vitro model of BBB, based on a three-cell-type co-culture system, in which rat neurons and astrocytes synergistically induce brain capillary endothelial cells to form a monolayer with permeability properties resembling those of the physiological BBB. Herein we report that the serum from patients with secondary progressive multiple sclerosis (SPMS) has a damaging effect on isolated neurons. This finding suggests that neuronal damaging in MS could be a primary event and not only secondary to myelin damage, as generally assumed. SPMS serum affects the permeability of the BBB model, as indicated by the decrease of the transendothelial electrical resistance (TEER). Moreover, as shown by both immunofluorescence and Western blot analyses, BBB breaking is accompanied by a decrease of the synthesis as well as the peripheral localization of occludin, a structural protein of the tight junctions that are responsible for BBB properties.
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