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Multiplexed, high-throughput measurements of cell contraction and endothelial barrier function
Vascular leak, protein exudation, and edema formation are events commonly triggered by inflammation and facilitated by gaps that form between adjacent endothelial cells (ECs) of the vasculature. In such paracellular gap formation, the role of EC contraction is widely implicated, and even therapeutic...
Autores principales: | , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
2018
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6309267/ https://www.ncbi.nlm.nih.gov/pubmed/30310180 http://dx.doi.org/10.1038/s41374-018-0136-2 |
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author | Rokhzan, Ramin Ghosh, Chandra C. Schaible, Niccole Notbohm, Jacob Yoshie, Haruka Ehrlicher, Allen J. Higgins, Sarah J. Zhang, Ruyuan Haller, Hermann Hardin, Corey C. David, Sascha Parikh, Samir M. Krishnan, Ramaswamy |
author_facet | Rokhzan, Ramin Ghosh, Chandra C. Schaible, Niccole Notbohm, Jacob Yoshie, Haruka Ehrlicher, Allen J. Higgins, Sarah J. Zhang, Ruyuan Haller, Hermann Hardin, Corey C. David, Sascha Parikh, Samir M. Krishnan, Ramaswamy |
author_sort | Rokhzan, Ramin |
collection | PubMed |
description | Vascular leak, protein exudation, and edema formation are events commonly triggered by inflammation and facilitated by gaps that form between adjacent endothelial cells (ECs) of the vasculature. In such paracellular gap formation, the role of EC contraction is widely implicated, and even therapeutically targeted. However, related measurement approaches remain slow, tedious, and complex to perform. Here, we have developed a multiplexed, high-throughput screen to simultaneously quantify paracellular gaps, EC contractile forces, and visualize F-actin stress fibers, and VE-Cadherin. As proof-of-principle, we examined barrier-protective mechanisms of the Rho-associated kinase inhibitor, Y-27632, and the canonical agonist of the Tie2 receptor, Angiopoietin-1 (Angpt-1). Y-27632 reduced EC contraction and actin stress fiber formation whereas Angpt-1 did not. Yet both agents reduced thrombin-, LPS-, and TNFα-induced paracellular gap formation. This unexpected result suggests that Angpt-1 can achieve barrier defense without reducing EC contraction, a mechanism that has not been previously described. This insight was enabled by the multiplex nature of the force-based platform. The high-throughput format we describe should accelerate both mechanistic studies and the screening of pharmacological modulators of endothelial barrier function. |
format | Online Article Text |
id | pubmed-6309267 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2018 |
record_format | MEDLINE/PubMed |
spelling | pubmed-63092672019-04-11 Multiplexed, high-throughput measurements of cell contraction and endothelial barrier function Rokhzan, Ramin Ghosh, Chandra C. Schaible, Niccole Notbohm, Jacob Yoshie, Haruka Ehrlicher, Allen J. Higgins, Sarah J. Zhang, Ruyuan Haller, Hermann Hardin, Corey C. David, Sascha Parikh, Samir M. Krishnan, Ramaswamy Lab Invest Article Vascular leak, protein exudation, and edema formation are events commonly triggered by inflammation and facilitated by gaps that form between adjacent endothelial cells (ECs) of the vasculature. In such paracellular gap formation, the role of EC contraction is widely implicated, and even therapeutically targeted. However, related measurement approaches remain slow, tedious, and complex to perform. Here, we have developed a multiplexed, high-throughput screen to simultaneously quantify paracellular gaps, EC contractile forces, and visualize F-actin stress fibers, and VE-Cadherin. As proof-of-principle, we examined barrier-protective mechanisms of the Rho-associated kinase inhibitor, Y-27632, and the canonical agonist of the Tie2 receptor, Angiopoietin-1 (Angpt-1). Y-27632 reduced EC contraction and actin stress fiber formation whereas Angpt-1 did not. Yet both agents reduced thrombin-, LPS-, and TNFα-induced paracellular gap formation. This unexpected result suggests that Angpt-1 can achieve barrier defense without reducing EC contraction, a mechanism that has not been previously described. This insight was enabled by the multiplex nature of the force-based platform. The high-throughput format we describe should accelerate both mechanistic studies and the screening of pharmacological modulators of endothelial barrier function. 2018-10-11 2019-01 /pmc/articles/PMC6309267/ /pubmed/30310180 http://dx.doi.org/10.1038/s41374-018-0136-2 Text en Users may view, print, copy, and download text and data-mine the content in such documents, for the purposes of academic research, subject always to the full Conditions of use:http://www.nature.com/authors/editorial_policies/license.html#terms |
spellingShingle | Article Rokhzan, Ramin Ghosh, Chandra C. Schaible, Niccole Notbohm, Jacob Yoshie, Haruka Ehrlicher, Allen J. Higgins, Sarah J. Zhang, Ruyuan Haller, Hermann Hardin, Corey C. David, Sascha Parikh, Samir M. Krishnan, Ramaswamy Multiplexed, high-throughput measurements of cell contraction and endothelial barrier function |
title | Multiplexed, high-throughput measurements of cell contraction and endothelial barrier function |
title_full | Multiplexed, high-throughput measurements of cell contraction and endothelial barrier function |
title_fullStr | Multiplexed, high-throughput measurements of cell contraction and endothelial barrier function |
title_full_unstemmed | Multiplexed, high-throughput measurements of cell contraction and endothelial barrier function |
title_short | Multiplexed, high-throughput measurements of cell contraction and endothelial barrier function |
title_sort | multiplexed, high-throughput measurements of cell contraction and endothelial barrier function |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6309267/ https://www.ncbi.nlm.nih.gov/pubmed/30310180 http://dx.doi.org/10.1038/s41374-018-0136-2 |
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