The ARP 2/3 complex mediates endothelial barrier function and recovery
Brown, Mary E.
Brinley, Brittany N.
Rizzo, Alicia N.
Garcia, Joe G.N.
Dudek, Steven M.
AffiliationUniv Arizona, Hlth Sci Ctr
MetadataShow full item record
PublisherSAGE PUBLICATIONS INC
CitationThe ARP 2/3 complex mediates endothelial barrier function and recovery 2017, 7 (1):200 Pulmonary Circulation
Rights© 2017 by Pulmonary Vascular Research Institute. Creative Commons Non Commercial CC-BY-NC: This article is distributed under the terms of the Creative Commons Attribution-NonCommercial 3.0 License.
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AbstractPulmonary endothelial cell (EC) barrier dysfunction and recovery is critical to the pathophysiology of acute respiratory distress syndrome. Cytoskeletal and subsequent cell membrane dynamics play a key mechanistic role in determination of EC barrier integrity. Here, we characterizAQe the actin related protein 2/3 (Arp 2/3) complex, a regulator of peripheral branched actin polymerization, in human pulmonary EC barrier function through studies of transendothelial electrical resistance (TER), intercellular gap formation, peripheral cytoskeletal structures and lamellipodia. Compared to control, Arp 2/3 inhibition with the small molecule inhibitor CK-666 results in a reduction of baseline barrier function (1,241 +/- 53 vs 988 +/- 64 ohm; p < 0.01), S1P-induced barrier enhancement and delayed recovery of barrier function after thrombin (143 +/- 14 vs 93 +/- 6 min; p < 0.01). Functional changes of Arp 2/3 inhibition on barrier integrity are associated temporally with increased intercellular gap area at baseline (0.456 +/- 0.02 vs 0.299 +/- 0.02; p < 0.05) and thirty minutes after thrombin (0.885 +/- 0.03 vs 0.754 +/- 0.03; p < 0.05). Immunofluorescent microscopy reveals reduced lamellipodia formation after S1P and during thrombin recovery in Arp 2/3 inhibited cells. Individual lamellipodia demonstrate reduced depth following Arp 2/3 inhibition vs vehicle at baseline (1.83 +/- 0.41 vs 2.55 +/- 0.46 mm; p < 0.05) and thirty minutes after S1P treatment (1.53 +/- 0.37 vs 2.09 +/- 0.36 mm; p < 0.05). These results establish a critical role for Arp 2/3 activity in determination of pulmonary endothelial barrier function and recovery through formation of EC lamellipodia and closure of intercellular gaps.
Note12 month embargo; First Published February 1, 2017
VersionFinal published version
SponsorsAmerican Lung Association [RG-349765]; National Institutes of Health [HL058064, HL088144]