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1 Chemical engineering, The Pennsylvania State University, University Park, PA, USA
2 Biomedical Engineering, The City College of The City University of New York, New York, NY, USA
3 Cellular and Molecular Physiology, Penn State College of Medicine, Hershey, PA, USA
4 Opthalmology, Penn State College of Medicine, Hershey, PA, USA
5 Cellular and Molecular Physiology, Penn State College of Medicine, Hershey, PA, USA; Opthalmology, Penn State College of Medicine, Hershey, PA, USA
* To whom correspondence should be addressed. E-mail: dantonetti{at}psu.edu.
A sudden increase in transmural pressure gradient across endothelial monolayers reduces hydraulic conductivity (Lp), a phenomenon known as the sealing effect. To further characterize this endothelial adaptive response, we measured bovine aortic endothelial cell (BAEC) permeability to albumin and 70 kDa dextran, Lp, and the solvent-drag reflection coefficients (
) during the sealing process. The diffusional permeability coefficients for albumin (Pdalb = 1.33 ± 0.18 x 10-6 cm/s) and dextran (Pddex = 0.60 ± 0.16 x 10-6 cm/s) were measured before pressure application. The effective permeabilities (measured when solvent drag contributes to solute transport) of albumin and dextran (Pealb and Pedex) were measured after application of a 10-cmH2O pressure gradient; during the first 2 h of pressure application, Pealb, Pedex, and Lp were significantly reduced by 2.0 ± 0.3-fold, 2.1 ± 0.3-fold, and 3.7 ± 0.3-fold, respectively. Immunostaining of the tight junction (TJ) protein zonula occludens-1 (ZO-1) was significantly increased at cell-cell contacts after application of transmural pressure. Cytochalasin D treatment significantly elevated transport but did not inhibit the adaptive response, whereas colchicine treatment had no effect on diffusive permeability but inhibited the adaptive response. Neither cytoskeletal inhibitor altered despite significantly elevating both Lp and Pe. Our data suggest that BAECs actively adapt to elevated transmural pressure by mobilizing ZO-1 to intercellular junctions via microtubules. A mechanical (passive) component of the sealing effect appears to reduce the size of a small pore system which allows the transport of water but not dextran or albumin. Furthermore, the structures of the TJ determine transport rates but do not define the selectivity of the monolayer to solutes ().
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