Impact of Na+ permeation on collective migration of pulmonary arterial endothelial cells.

Collective migration of endothelial cells is important for wound healing and angiogenesis. During such migration, each constituent endothelial cell coordinates its magnitude and direction of migration with its neighbors while retaining intercellular adhesion. Ensuring coordination and cohesion invol...

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Autores principales:	Ningyong Xu, Linn Ayers, Viktoriya Pastukh, Mikhail Alexeyev, Troy Stevens, Dhananjay T Tambe
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/236f15db273a4cc98e7bc0419fc5a591
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spelling	oai:doaj.org-article:236f15db273a4cc98e7bc0419fc5a5912021-12-02T20:05:43ZImpact of Na+ permeation on collective migration of pulmonary arterial endothelial cells.1932-620310.1371/journal.pone.0250095https://doaj.org/article/236f15db273a4cc98e7bc0419fc5a5912021-01-01T00:00:00Zhttps://doi.org/10.1371/journal.pone.0250095https://doaj.org/toc/1932-6203Collective migration of endothelial cells is important for wound healing and angiogenesis. During such migration, each constituent endothelial cell coordinates its magnitude and direction of migration with its neighbors while retaining intercellular adhesion. Ensuring coordination and cohesion involves a variety of intra- and inter-cellular signaling processes. However, the role of permeation of extracellular Na+ in collective cell migration remains unclear. Here, we examined the effect of Na+ permeation in collective migration of pulmonary artery endothelial cell (PAEC) monolayers triggered by either a scratch injury or a barrier removal over 24 hours. In the scratch assay, PAEC monolayers migrated in two approximately linear phases. In the first phase, wound closure started with fast speed which then rapidly reduced within 5 hours after scratching. In the second phase, wound closure maintained at slow and stable speed from 6 to 24 hours. In the absence of extracellular Na+, the wound closure distance was reduced by >50%. Fewer cells at the leading edge protruded prominent lamellipodia. Beside transient gaps, some sustained interendothelial gaps also formed and progressively increased in size over time, and some fused with adjacent gaps. In the absence of both Na+ and scratch injury, PAEC monolayer migrated even more slowly, and interendothelial gaps obviously increased in size towards the end. Pharmacological inhibition of the epithelial Na+ channel (ENaC) using amiloride reduced wound closure distance by 30%. Inhibition of both the ENaC and the Na+/Ca2+ exchanger (NCX) using benzamil further reduced wound closure distance in the second phase and caused accumulation of floating particles in the media. Surprisingly, pharmacological inhibition of the Ca2+ release-activated Ca2+ (CRAC) channel protein 1 (Orai1) using GSK-7975A, the transient receptor potential channel protein 1 and 4 (TRPC1/4) using Pico145, or both Orai1 and TRPC1/4 using combined GSK-7975A and Pico145 treatment did not affect wound closure distance dramatically. Nevertheless, the combined treatment appeared to cause accumulation of floating particles. Note that GSK-7975A also inhibits small inward Ca2+ currents via Orai2 and Orai3 channels, whereas Pico145 also blocks TRPC4, TRPC5, and TRPC1/5 channels. By contrast, gene silence of Orai1 by shRNAs led to a 25% reduction of wound closure in the first 6 hours but had no effect afterwards. However, in the absence of extracellular Na+ or cellular injury, Orai1 did not affect PAEC collective migration. Overall, the data reveal that Na+ permeation into cells contributes to PAEC monolayer collective migration by increasing lamellipodial formation, reducing accumulation of floating particles, and improving intercellular adhesion.Ningyong XuLinn AyersViktoriya PastukhMikhail AlexeyevTroy StevensDhananjay T TambePublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 16, Iss 4, p e0250095 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Ningyong Xu Linn Ayers Viktoriya Pastukh Mikhail Alexeyev Troy Stevens Dhananjay T Tambe Impact of Na+ permeation on collective migration of pulmonary arterial endothelial cells.
description	Collective migration of endothelial cells is important for wound healing and angiogenesis. During such migration, each constituent endothelial cell coordinates its magnitude and direction of migration with its neighbors while retaining intercellular adhesion. Ensuring coordination and cohesion involves a variety of intra- and inter-cellular signaling processes. However, the role of permeation of extracellular Na+ in collective cell migration remains unclear. Here, we examined the effect of Na+ permeation in collective migration of pulmonary artery endothelial cell (PAEC) monolayers triggered by either a scratch injury or a barrier removal over 24 hours. In the scratch assay, PAEC monolayers migrated in two approximately linear phases. In the first phase, wound closure started with fast speed which then rapidly reduced within 5 hours after scratching. In the second phase, wound closure maintained at slow and stable speed from 6 to 24 hours. In the absence of extracellular Na+, the wound closure distance was reduced by >50%. Fewer cells at the leading edge protruded prominent lamellipodia. Beside transient gaps, some sustained interendothelial gaps also formed and progressively increased in size over time, and some fused with adjacent gaps. In the absence of both Na+ and scratch injury, PAEC monolayer migrated even more slowly, and interendothelial gaps obviously increased in size towards the end. Pharmacological inhibition of the epithelial Na+ channel (ENaC) using amiloride reduced wound closure distance by 30%. Inhibition of both the ENaC and the Na+/Ca2+ exchanger (NCX) using benzamil further reduced wound closure distance in the second phase and caused accumulation of floating particles in the media. Surprisingly, pharmacological inhibition of the Ca2+ release-activated Ca2+ (CRAC) channel protein 1 (Orai1) using GSK-7975A, the transient receptor potential channel protein 1 and 4 (TRPC1/4) using Pico145, or both Orai1 and TRPC1/4 using combined GSK-7975A and Pico145 treatment did not affect wound closure distance dramatically. Nevertheless, the combined treatment appeared to cause accumulation of floating particles. Note that GSK-7975A also inhibits small inward Ca2+ currents via Orai2 and Orai3 channels, whereas Pico145 also blocks TRPC4, TRPC5, and TRPC1/5 channels. By contrast, gene silence of Orai1 by shRNAs led to a 25% reduction of wound closure in the first 6 hours but had no effect afterwards. However, in the absence of extracellular Na+ or cellular injury, Orai1 did not affect PAEC collective migration. Overall, the data reveal that Na+ permeation into cells contributes to PAEC monolayer collective migration by increasing lamellipodial formation, reducing accumulation of floating particles, and improving intercellular adhesion.
format	article
author	Ningyong Xu Linn Ayers Viktoriya Pastukh Mikhail Alexeyev Troy Stevens Dhananjay T Tambe
author_facet	Ningyong Xu Linn Ayers Viktoriya Pastukh Mikhail Alexeyev Troy Stevens Dhananjay T Tambe
author_sort	Ningyong Xu
title	Impact of Na+ permeation on collective migration of pulmonary arterial endothelial cells.
title_short	Impact of Na+ permeation on collective migration of pulmonary arterial endothelial cells.
title_full	Impact of Na+ permeation on collective migration of pulmonary arterial endothelial cells.
title_fullStr	Impact of Na+ permeation on collective migration of pulmonary arterial endothelial cells.
title_full_unstemmed	Impact of Na+ permeation on collective migration of pulmonary arterial endothelial cells.
title_sort	impact of na+ permeation on collective migration of pulmonary arterial endothelial cells.
publisher	Public Library of Science (PLoS)
publishDate	2021
url	https://doaj.org/article/236f15db273a4cc98e7bc0419fc5a591
work_keys_str_mv	AT ningyongxu impactofnapermeationoncollectivemigrationofpulmonaryarterialendothelialcells AT linnayers impactofnapermeationoncollectivemigrationofpulmonaryarterialendothelialcells AT viktoriyapastukh impactofnapermeationoncollectivemigrationofpulmonaryarterialendothelialcells AT mikhailalexeyev impactofnapermeationoncollectivemigrationofpulmonaryarterialendothelialcells AT troystevens impactofnapermeationoncollectivemigrationofpulmonaryarterialendothelialcells AT dhananjayttambe impactofnapermeationoncollectivemigrationofpulmonaryarterialendothelialcells
_version_	1718375409916051456

Impact of Na+ permeation on collective migration of pulmonary arterial endothelial cells.

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