Endothelial cells form a continuous surface that communicates with all parts of the body, yet they exhibit functional and structural specializations in various segments and locations of the vasculature. For instance, endothelial cells that line the capillaries of the brain are relatively thick, exhbit low permeability to macromolecular and high electrical resistance. Endothelial cells that line the capillaries of the intestine are thin and atenuated, have high permeability to macromolecules, and low electrical resistance. Even within a single organ endothelical cell specialization is evident. Arteriolar and true capillary endothelial cells lack histamine receptors while post-capillary venules possess these receptors. Most importantly, from the standpoint of inflammation, post capillary venules are the sites of leukocyte emigration. The research outlined in this proposal is aimed at studying the surface properties of endothelial cells and the role(s) of their surfaces in regulating leukocyte emigration. We describe a method for growing endothelial cells on amniotic membrane such that the endothelial cell monolayers exhibit silver stained intercellular junctions, are impermeable to macromolecular tracers, and show resistance to the passage of electrical current. We believe these cultures are excellent models of vessel walls. The studies proposed will use this """"""""model"""""""" vessel wall to investigate the structure of the surfaces of endothelial cells derived from different anatomic locations, alterations in the surface properties of endothelial cells cultured on natural (amnion) and artificial (plastic) substrates, the structure of endothelial cell intercellular junctions, and the mechanisms by which leukocytes open these junctions and emigrate into the extravascular compartment.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
5R01HL032210-05
Application #
3343533
Study Section
Pathology A Study Section (PTHA)
Project Start
1984-07-01
Project End
1989-06-30
Budget Start
1988-07-01
Budget End
1989-06-30
Support Year
5
Fiscal Year
1988
Total Cost
Indirect Cost
Name
Columbia University (N.Y.)
Department
Type
Schools of Medicine
DUNS #
064931884
City
New York
State
NY
Country
United States
Zip Code
10027
Hixenbaugh, E A; Goeckeler, Z M; Papaiya, N N et al. (1997) Stimulated neutrophils induce myosin light chain phosphorylation and isometric tension in endothelial cells. Am J Physiol 273:H981-8
Loike, J D; Hickman, S; Kuang, K et al. (1996) Sodium-glucose cotransporters display sodium- and phlorizin-dependent water permeability. Am J Physiol 271:C1774-9
Loike, J D; el Khoury, J; Cao, L et al. (1995) Fibrin regulates neutrophil migration in response to interleukin 8, leukotriene B4, tumor necrosis factor, and formyl-methionyl-leucyl-phenylalanine. J Exp Med 181:1763-72
el Khoury, J; Thomas, C A; Loike, J D et al. (1994) Macrophages adhere to glucose-modified basement membrane collagen IV via their scavenger receptors. J Biol Chem 269:10197-200
Heltianu, C; Dobrila, L; Stanescu, M et al. (1994) A method for selective radiolabeling of lung endothelium plasmalemmal vesicles, in situ. Eur J Cell Biol 64:61-70
Huang, A J; Manning, J E; Bandak, T M et al. (1993) Endothelial cell cytosolic free calcium regulates neutrophil migration across monolayers of endothelial cells. J Cell Biol 120:1371-80
Loike, J D; Cao, L; Brett, J et al. (1992) Hypoxia induces glucose transporter expression in endothelial cells. Am J Physiol 263:C326-33
Ogawa, S; Koga, S; Kuwabara, K et al. (1992) Hypoxia-induced increased permeability of endothelial monolayers occurs through lowering of cellular cAMP levels. Am J Physiol 262:C546-54
Huang, A J; Silverstein, S C; Malawista, S E (1991) Cryopreserved cytoplasts from human neutrophils migrate across monolayers of human endothelial cells in response to a chemoattractant gradient. J Leukoc Biol 50:624-7
Luckett, P M; Fischbarg, J; Bhattacharya, J et al. (1989) Hydraulic conductivity of endothelial cell monolayers cultured on human amnion. Am J Physiol 256:H1675-83

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