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PMID: 1518814 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Electrical method for detection of endothelial cell shape change in real time: assessment of endothelial barrier function.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 89 ·No. 17 ·1992-09-01 ·Pages 7919-23

Tiruppathi C, Malik AB, Del Vecchio PJ, Keese CR, Giaever I

Abstract

We have developed an electrical method to study endothelial cell shape changes in real time in order to examine the mechanisms of alterations in the endothelial barrier function. Endothelial shape changes were quantified by using a monolayer of endothelial cells grown on a small (10(-3) cm2) evaporated gold electrode and measuring the changes in electrical impedance. Bovine pulmonary microvessel endothelial cells and bovine pulmonary artery endothelial cells were used to study the effects of alpha-thrombin on cell-shape dynamics by the impedance measurement. alpha-Thrombin produced a dose-dependent decrease in impedance that occurred within 0.5 min in both cell types, indicative of retraction of endothelial cells and widening of interendothelial junctions because of "rounding up" of the cells. The alpha-thrombin-induced decrease in impedance persisted for approximately 2 hr, after which the value recovered to basal levels. Pretreatment of endothelial cells with the protein kinase C inhibitor, calphostin C, or with 8-bromoadenosine 3',5'-cyclic monophosphate prevented the decreased impedance, suggesting that the endothelial cell change is modulated by activation of second-messenger pathways. The alpha-thrombin-induced decrease in impedance was in agreement with the previously observed increases in transendothelial albumin permeability and evidence of formation of intercellular gaps after alpha-thrombin challenge. The impedance measurement may be a valuable in vitro method for the assessment of mechanisms of decreased endothelial barrier function occurring with inflammatory mediators. Since the rapidly occurring changes in endothelial cell shape in response to mediators such as thrombin are mediated activation of second-messenger pathways, the ability to monitor endothelial cell dynamics in real time may provide insights into the signal-transduction events mediating the increased endothelial permeability.

MeSH Terms
Animals Cattle Cells, Cultured Electrophysiology Endothelium, Vascular/cytology,physiology In Vitro Techniques Membrane Potentials Permeability Thrombin/pharmacology
Chemicals
Thrombin
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Tiruppathi C
Department of Physiology and Cell Biology, Albany Medical College, NY 12208.
Malik A B
Del Vecchio P J
Keese C R
Giaever I
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34 references, click to expand
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.89.17.7919
Published
1992-09-01
Pages
7919-23
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC49826
Subset
IM
Grants
NHLBI NIH HHS · HL27016 · United States
NHLBI NIH HHS · HL45638 · United States
NHLBI NIH HHS · HL46350 · United States
External Links
PubMed source DOI: 10.1073/pnas.89.17.7919 PMC Full Text View journal details
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