Hyperoxia increases the elastic modulus of alveolar epithelial cells through Rho kinase

Kristina R. Wilhelm, Esra Roan, Manik C. Ghosh, Kaushik Parthasarathi, Christopher Waters

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

Patients with acute lung injury are administered high concentrations of oxygen during mechanical ventilation, and while both hyperoxia and mechanical ventilation are necessary, each can independently cause additional injury. However, the precise mechanisms that lead to injury are not well understood. We hypothesized that alveolar epithelial cells may be more susceptible to injury caused by mechanical ventilation because hyperoxia causes cells to be stiffer due to increased filamentous actin (f-actin) formation via the GTPase RhoA and its effecter Rho kinase (ROCK). We examined cytoskeletal structures in cultured murine lung alveolar epithelial cells (MLE-12) under normoxic and hyperoxic (48 h) conditions. We also measured cell elasticity (E) using an atomic force microscope in the indenter mode. Hyperoxia caused increased f-actin stress fibers and bundle formation, an increase in g- and f-actin, an increase in nuclear area and a decrease in nuclear height, and cells became stiffer (higher E). Treatment with an inhibitor (Y-27632) of ROCK significantly decreased E and prevented the cytoskeletal changes, while it did not influence the nuclear height and area. Pre-exposure of cells to hyperoxia promoted detachment when cells were subsequently stretched cyclically, but the ROCK inhibitor prevented this effect. Hyperoxia caused thickening of vinculin focal adhesion plaques, and inhibition of ROCK reduced the formation of distinct focal adhesion plaques. Phosphorylation of focal adhesion kinase was significantly reduced by both hyperoxia and treatment with Y-27632. Hyperoxia caused increased cell stiffness and promoted cell detachment during stretch. These effects were ameliorated by inhibition of ROCK.

Original languageEnglish (US)
Pages (from-to)957-969
Number of pages13
JournalFEBS Journal
Volume281
Issue number3
DOIs
StatePublished - Jan 1 2014

Fingerprint

Alveolar Epithelial Cells
rho-Associated Kinases
Hyperoxia
Elastic Modulus
Actins
Elastic moduli
Focal Adhesions
Adhesion
Vinculin
Focal Adhesion Protein-Tyrosine Kinases
Artificial Respiration
Phosphorylation
Maximum likelihood estimation
GTP Phosphohydrolases
Elasticity
Microscopes
Stiffness
Wounds and Injuries
Oxygen
Fibers

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Hyperoxia increases the elastic modulus of alveolar epithelial cells through Rho kinase. / Wilhelm, Kristina R.; Roan, Esra; Ghosh, Manik C.; Parthasarathi, Kaushik; Waters, Christopher.

In: FEBS Journal, Vol. 281, No. 3, 01.01.2014, p. 957-969.

Research output: Contribution to journalArticle

Wilhelm, Kristina R. ; Roan, Esra ; Ghosh, Manik C. ; Parthasarathi, Kaushik ; Waters, Christopher. / Hyperoxia increases the elastic modulus of alveolar epithelial cells through Rho kinase. In: FEBS Journal. 2014 ; Vol. 281, No. 3. pp. 957-969.
@article{6330bcf94cca4a36bb8e35fd49708371,
title = "Hyperoxia increases the elastic modulus of alveolar epithelial cells through Rho kinase",
abstract = "Patients with acute lung injury are administered high concentrations of oxygen during mechanical ventilation, and while both hyperoxia and mechanical ventilation are necessary, each can independently cause additional injury. However, the precise mechanisms that lead to injury are not well understood. We hypothesized that alveolar epithelial cells may be more susceptible to injury caused by mechanical ventilation because hyperoxia causes cells to be stiffer due to increased filamentous actin (f-actin) formation via the GTPase RhoA and its effecter Rho kinase (ROCK). We examined cytoskeletal structures in cultured murine lung alveolar epithelial cells (MLE-12) under normoxic and hyperoxic (48 h) conditions. We also measured cell elasticity (E) using an atomic force microscope in the indenter mode. Hyperoxia caused increased f-actin stress fibers and bundle formation, an increase in g- and f-actin, an increase in nuclear area and a decrease in nuclear height, and cells became stiffer (higher E). Treatment with an inhibitor (Y-27632) of ROCK significantly decreased E and prevented the cytoskeletal changes, while it did not influence the nuclear height and area. Pre-exposure of cells to hyperoxia promoted detachment when cells were subsequently stretched cyclically, but the ROCK inhibitor prevented this effect. Hyperoxia caused thickening of vinculin focal adhesion plaques, and inhibition of ROCK reduced the formation of distinct focal adhesion plaques. Phosphorylation of focal adhesion kinase was significantly reduced by both hyperoxia and treatment with Y-27632. Hyperoxia caused increased cell stiffness and promoted cell detachment during stretch. These effects were ameliorated by inhibition of ROCK.",
author = "Wilhelm, {Kristina R.} and Esra Roan and Ghosh, {Manik C.} and Kaushik Parthasarathi and Christopher Waters",
year = "2014",
month = "1",
day = "1",
doi = "10.1111/febs.12661",
language = "English (US)",
volume = "281",
pages = "957--969",
journal = "FEBS Journal",
issn = "1742-464X",
publisher = "Wiley-Blackwell",
number = "3",

}

TY - JOUR

T1 - Hyperoxia increases the elastic modulus of alveolar epithelial cells through Rho kinase

AU - Wilhelm, Kristina R.

AU - Roan, Esra

AU - Ghosh, Manik C.

AU - Parthasarathi, Kaushik

AU - Waters, Christopher

PY - 2014/1/1

Y1 - 2014/1/1

N2 - Patients with acute lung injury are administered high concentrations of oxygen during mechanical ventilation, and while both hyperoxia and mechanical ventilation are necessary, each can independently cause additional injury. However, the precise mechanisms that lead to injury are not well understood. We hypothesized that alveolar epithelial cells may be more susceptible to injury caused by mechanical ventilation because hyperoxia causes cells to be stiffer due to increased filamentous actin (f-actin) formation via the GTPase RhoA and its effecter Rho kinase (ROCK). We examined cytoskeletal structures in cultured murine lung alveolar epithelial cells (MLE-12) under normoxic and hyperoxic (48 h) conditions. We also measured cell elasticity (E) using an atomic force microscope in the indenter mode. Hyperoxia caused increased f-actin stress fibers and bundle formation, an increase in g- and f-actin, an increase in nuclear area and a decrease in nuclear height, and cells became stiffer (higher E). Treatment with an inhibitor (Y-27632) of ROCK significantly decreased E and prevented the cytoskeletal changes, while it did not influence the nuclear height and area. Pre-exposure of cells to hyperoxia promoted detachment when cells were subsequently stretched cyclically, but the ROCK inhibitor prevented this effect. Hyperoxia caused thickening of vinculin focal adhesion plaques, and inhibition of ROCK reduced the formation of distinct focal adhesion plaques. Phosphorylation of focal adhesion kinase was significantly reduced by both hyperoxia and treatment with Y-27632. Hyperoxia caused increased cell stiffness and promoted cell detachment during stretch. These effects were ameliorated by inhibition of ROCK.

AB - Patients with acute lung injury are administered high concentrations of oxygen during mechanical ventilation, and while both hyperoxia and mechanical ventilation are necessary, each can independently cause additional injury. However, the precise mechanisms that lead to injury are not well understood. We hypothesized that alveolar epithelial cells may be more susceptible to injury caused by mechanical ventilation because hyperoxia causes cells to be stiffer due to increased filamentous actin (f-actin) formation via the GTPase RhoA and its effecter Rho kinase (ROCK). We examined cytoskeletal structures in cultured murine lung alveolar epithelial cells (MLE-12) under normoxic and hyperoxic (48 h) conditions. We also measured cell elasticity (E) using an atomic force microscope in the indenter mode. Hyperoxia caused increased f-actin stress fibers and bundle formation, an increase in g- and f-actin, an increase in nuclear area and a decrease in nuclear height, and cells became stiffer (higher E). Treatment with an inhibitor (Y-27632) of ROCK significantly decreased E and prevented the cytoskeletal changes, while it did not influence the nuclear height and area. Pre-exposure of cells to hyperoxia promoted detachment when cells were subsequently stretched cyclically, but the ROCK inhibitor prevented this effect. Hyperoxia caused thickening of vinculin focal adhesion plaques, and inhibition of ROCK reduced the formation of distinct focal adhesion plaques. Phosphorylation of focal adhesion kinase was significantly reduced by both hyperoxia and treatment with Y-27632. Hyperoxia caused increased cell stiffness and promoted cell detachment during stretch. These effects were ameliorated by inhibition of ROCK.

UR - http://www.scopus.com/inward/record.url?scp=84897071312&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84897071312&partnerID=8YFLogxK

U2 - 10.1111/febs.12661

DO - 10.1111/febs.12661

M3 - Article

VL - 281

SP - 957

EP - 969

JO - FEBS Journal

JF - FEBS Journal

SN - 1742-464X

IS - 3

ER -