Histamine alters endothelial barrier function at cell-cell and cell-matrix sites.
To determine how histamine regulates endothelial barrier function through an integrative cytoskeletal network, we mathematically modeled the resistance across an endothelial cell-covered electrode as a function of cell-cell, cell-matrix, and transcellular resistances. Based on this approach, histamine initiated a rapid decrease in transendothelial resistance predominantly through decreases in cell-cell resistance in confluent cultured human umbilical vein endothelial cells (HUVECs). Restoration of resistance was characterized by initially increasing cell-matrix resistance, with later increases in cell-cell resistance. Thus histamine disrupts barrier function by specifically disrupting cell-cell adhesion and restores barrier function in part through direct effects on cell-matrix adhesion. To validate the precision of our technique, histamine increased the resistance in subconfluent HUVECs in which there was no cell-cell contact. Exposure of confluent monolayers to an antibody against cadherin-5 caused a predominant decrease in cell-cell resistance, whereas the resistance was unaffected by the antibody to cadherin-5 in subconfluent cells. Furthermore, we observed an increase predominantly in cell-cell resistance in ECV304 cells that were transfected with a plasmid containing a glucocorticoid-inducible promoter controlling expression of E-cadherin. Transmission electron microscopy confirmed tens of nanometer displacements between adjacent cells at a time point in which histamine maximally decreased cell-cell resistance.
Moy
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AB
Department of Internal Medicine, University of Iowa, Iowa City, Iowa 52242, USA. alan-moy@uiowa.edu
Winter
M
M
Kamath
A
A
Blackwell
K
K
Reyes
G
G
Giaever
I
I
Keese
C
C
Shasby
D M
DM
D015820:Cadherins / Q000032:analysis / Q000235:genetics; D002448:Cell Adhesion / Q000187:drug effects; D002450:Cell Communication / Q000187:drug effects* / Q000502:physiology; D002478:Cells, Cultured; D017097:Electric Impedance; D004847:Epithelial Cells / Q000737:chemistry / Q000187:drug effects* / Q000648:ultrastructure*; D005109:Extracellular Matrix / Q000187:drug effects / Q000502:physiology; D005786:Gene Expression Regulation / Q000502:physiology; D006632:Histamine / Q000494:pharmacology*; D006801:Humans; D007365:Intercellular Junctions / Q000737:chemistry / Q000187:drug effects* / Q000648:ultrastructure; D008839:Microelectrodes; D008854:Microscopy, Electron; D008954:Models, Biological; D011401:Promoter Regions, Genetic / Q000502:physiology; D014162:Transfection; D014471:Umbilical Veins / Q000166:cytology
D015820:Cadherins; D006632:Histamine