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Three‐dimensional Monolayer Stresses of Neutrophil Diapedesis
Author(s) -
Yeh YiTing,
Serrano Ricardo,
Li YiShuan,
Alamo Juan Carlos,
Chien Shu,
Lasheras Juan Carlos
Publication year - 2016
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.30.1_supplement.723.12
Subject(s) - adherens junction , microbiology and biotechnology , endothelium , biophysics , endothelial stem cell , cell junction , monolayer , tight junction , chemistry , inflammation , cell , biology , immunology , cadherin , biochemistry , in vitro , endocrinology
Neutrophil extravasation is a key step for immune defense and inflammation. Upon activation, neutrophils adhere firmly to the vascular endothelial cells (ECs), breach cell‐cell junctions, transmigrate across endothelium, and cross the basement membrane (BM) in the process of diapedesis. The biochemical mechanisms for diapedesis at EC junctional sites are well defined, but the biomechanical mechanisms remain largely undefined. The movement of neutrophils across the endothelium junction involves 3‐dimensional (3‐D) interactions, and hence it is important to understand the 3‐D cellular forces generated by neutrophil‐EC interaction in order to comprehend the biomechanical regulation of the diapedesis process. We hypothesize that the neutrophil‐EC interaction induces 3‐D cellular stresses to breach the cell junction, and serves as a crucial determinant for diapedesis. We have developed a 3‐D monolayer stress microscopy method (3‐D MSM) to measure and analyze the spatiotemporal evolution of 3‐D monolayer stresses during diapedesis. Our results show significant 3‐D deformations of the EC monolayer during neutrophil diapedesis that induce a mechanical disruption of the cell‐cell junctions, quantified with 3‐D MSM. The increase in barrier function induced by Angiopoietin‐1 and FGF treatments was associated with an augmentation of 3‐D displacements during diapedesis, whereas the negative modulation of junction integrity by thrombin treatment resulted in the reduction of 3‐D displacements. These results indicate that alterations of EC junction integrity lead to changes of diapedesis efficiency. By using the knockdown technique, we identified the adherens junction protein VE‐Cadherin as a key regulator for modulating EC junction integrity and the 3‐D displacements during diapedesis. In summary, we have developed a new approach to study mechanical regulation during diapedesis and provided new insights into the biomechanical mechanisms that control mechanical stress dynamics between neutrophils and the endothelium junction.