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A DYNAMIC MODEL OF THE BLOOD-BRAIN BARRIER
Abstract number: P-S-414
Colgan1 O.C., Cummins2 P.M., Kerrigan1 S.W., Cox1 D.
11Molecular and Cellular Therapeutics, Royal College of Surgeons in Ireland 22Vascular Health Research Centre, Dublin City University, Dublin, Ireland
How-to-cite Colgan OC, Cummins PM, Kerrigan SW, Cox D. A DYNAMIC MODEL OF THE BLOOD-BRAIN BARRIER. J Thromb Haemost 2007; 5 Supplement 2: P-S-414
Abstract
Introduction: In vivo interactions between endothelial cells (ECs) and blood-borne componenets (i.e. platelets, red blood cells, bacteria etc.) occur under dynamic conditions due to shear forces caused by blood flowing through vessels. ECs in vitro are frequently cultured under static conditions, however, several studies have shown that ECs exposed to shear express a phenotype, which more closely reflects the in vivo situation than their unsheared counterparts. In the microvasculature, such as that of the blood-brain barrier (BBB), shear is the primary haemodynamic force. Tight junctions (TJs) between adjacent ECs give rise to the BBB with expression and localization of Occludin (Occ) and ZO-1 being integral to TJ formation and function. We sought to elucidate if exposure Bovine Brain Microvascular Cells (BBMvECs) to shear results in the expression of EC phenotype similar to in vivo situation.
Methods: Confluent BBMvEC on 6-well plates were subjected to 10 dynes/cm2 non-pulsatile laminar shear for 24 h using an orbital shaker. Cells were harvested for Western blot/Real-Time PCR and immunocytochemical analysis. ZO-1 and Occ were immunoprecipitated to determine their co-association. EC barrier was monitored by tracer studies using FD40 FITC labeled dextran.
Results: Following shear, Occ and ZO-1 protein levels increased to 1.70.2 and 1.20.1 fold respectively, concomitant with a significant increase in mRNA (Occ 2.70.3; ZO-1 1.30.0 fold) and a significant increase in Occ/ZO-1 association (1.90.1 fold). After 24 h, f-actin realigned in the direction of shear, accompanied by increased localization of Occ and ZO-1 to the membrane. EC permeability to FITC dextran decreased by 622% following shear.
Conclusions: These results clearly suggest that shear regulates occludin and ZO-1 expression and localization, and by definition, barrier function in BBMvEC. This dynamic model will provide a system to study the BBB and the pathogenesis of neurological disorders.
To cite this abstract use the following format:
Journal of Thrombosis and Haemostasis 2007; Volume 5, Supplement 2: abstract number
Session Details
| Date: |
01/08/2007
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| Time: |
00:00-00:00
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| Session name: |
XXIst ISTH Congress |
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| Location: |
Oxford, UK |
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