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Polarized Actin Structural Dynamics in Response to Cyclic Uniaxial Stretch

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Abstract

Endothelial cell (EC) alignment to directional flow or stretch supports anti-inflammatory functions, but mechanisms controlling polarized structural adaptation in response to physical cues remain unclear. This study aimed to determine whether factors associated with early actin edge ruffling implicated in cell polarization are prerequisite for stress fiber (SF) reorientation in response to cyclic uniaxial stretch. Time-lapse analysis of EGFP-actin in confluent ECs showed that onset of either cyclic uniaxial or equibiaxial stretch caused a non-directional increase in edge ruffling. Edge activity was concentrated in a direction perpendicular to the stretch axis after 60 min, consistent with the direction of SF alignment. Rho-kinase inhibition caused reorientation of both stretch-induced edge ruffling and SF alignment parallel to the stretch axis. Arp2/3 inhibition attenuated stretch-induced cell elongation and disrupted polarized edge dynamics and microtubule organizing center reorientation, but it had no effect on the extent of SF reorientation. Disrupting localization of p21-activated kinase did not prevent stretch-induced SF reorientation, suggesting that this Rac effector is not critical in regulating stretch-induced cytoskeletal remodeling. Overall, these results suggest that directional edge ruffling is not a primary mechanism that guides SF reorientation in response to stretch; the two events are coincident but not causal.

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Abbreviations

EC:

Endothelial cell

SF:

Stress fiber

Arp2/3:

Actin-related protein-2/3

MTOC:

Microtubule organizing center

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Acknowledgements

The authors thank A. Wayne Orr for the generous gift of Nck-binding PAK peptides and Martin Schwartz for valuable discussions. Supported by NIH Grant HL080956 and by a University of Virginia VPRGS Award.

Conflict of interest

Lawrence Huang and Brian P. Helmke declare that they have no conflicts of interest.

Statements of Human and Animal Rights and Informed Consent

No human studies were carried out by the authors for this article. No animal studies were carried out by the authors for this article.

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Author notes

  1. Lawrence Huang

    Present address: Department of Surgery, University of California, San Francisco, San Francisco, CA, 94143, USA

Authors and Affiliations

  1. Department of Biomedical Engineering, University of Virginia, P. O. Box 800759, Charlottesville, VA, 22908, USA

    Lawrence Huang & Brian P. Helmke

  2. Robert M. Berne Cardiovascular Research Center, University of Virginia, P. O. Box 800759, Charlottesville, VA, 22908, USA

    Brian P. Helmke

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  1. Lawrence Huang
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Correspondence to Brian P. Helmke.

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Associate Editor Michael R. King oversaw the review of this article.

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Huang, L., Helmke, B.P. Polarized Actin Structural Dynamics in Response to Cyclic Uniaxial Stretch. Cel. Mol. Bioeng. 8, 160–177 (2015). https://doi.org/10.1007/s12195-014-0370-7

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