High precision measurement of electrical resistance across endothelial cell monolayers.
Effects of vasoactive agonists on endothelial permeability was assessed by measurement of transendothelial electrical resistance (TEER) of human umbilical vein endothelial cells (HUVECs) grown on porous polycarbonate supports. Because of the low values of TEER obtained in this preparation (< 5 omega cm2) a design of an Ussing type recording chamber was chosen that provided for a homogeneous electric field across the monolayer and for proper correction of series resistances. Precision current pulses and appropriate rates of sampling and averaging of the voltage signal allowed for measurement of < 0.1 omega resistance changes of the endothelium on top of a 21 omega series resistance of the support and bathing fluid layers. Histamine (10 microM) and thrombin (10 U/ml) induced an abrupt and substantial decrease of TEER, bradykinin (1 microM) was less effective, PAF (380 nM) and LTC4 (1 microM) had no effect. TEER was also reduced by the calcium ionophore A-23187 (10 microM). The technique allows for measurements of TEER in low resistance monolayer cultures with high precision and time resolution. The results obtained extend previous observations in providing quantitative data on the increase of permeability of HUVECs in response to vasoactive agonists.
Tschugguel
W
W
Department of General and Experimental Pathology, University of Vienna School of Medicine, Austria.
Zhegu
Z
Z
Gajdzik
L
L
Maier
M
M
Binder
B R
BR
Graf
J
J
D001920:Bradykinin / Q000494:pharmacology; D000001:Calcimycin / Q000494:pharmacology; D002478:Cells, Cultured; D017097:Electric Impedance; D004730:Endothelium, Vascular / Q000187:drug effects / Q000502:physiology*; D006632:Histamine / Q000494:pharmacology; D006801:Humans; D017997:Leukotriene C4 / Q000494:pharmacology; D010972:Platelet Activating Factor / Q000494:pharmacology; D013917:Thrombin / Q000494:pharmacology; D014471:Umbilical Veins / Q000166:cytology / Q000187:drug effects / Q000502:physiology
D010972:Platelet Activating Factor; D017997:Leukotriene C4; D000001:Calcimycin; D006632:Histamine; D013917:Thrombin; D001920:Bradykinin