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Endothelial Cell Membrane Sensitivity to Shear Stress is Lipid Domain Dependent

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Abstract

Blood flow-associated shear stress causes physiological and pathophysiological biochemical processes in endothelial cells that may be initiated by alterations in plasma membrane lipid domains characterized as liquid-ordered (lo), such as rafts or caveolae, or liquid-disordered (ld). To test for domain–dependent shear sensitivity, we used time-correlated single photon counting instrumentation to assess the photophysics and dynamics of the domain-selective lipid analogues DiI-C12 and DiI-C18 in endothelial cells subjected to physiological fluid shear stress. Under static conditions, DiI-C12 fluorescence lifetime was less than DiI-C18 lifetime and the diffusion coefficient of DiI-C12 was greater than the DiI-C18 diffusion coefficient, confirming that DiI-C12 probes ld, a more fluid membrane environment, and DiI-C18 probes the lo phase. Domains probed by DiI-C12 exhibited an early (10 s) and transient decrease of fluorescence lifetime after the onset of shear while domains probed by DiI-C18 exhibited a delayed decrease of fluorescence lifetime that was sustained for the 2 min the cells were subjected to flow. The diffusion coefficient of DiI-C18 increased after shear imposition, while that of DiI-C12 remained constant. Determination of the number of molecules (N) in the control volume suggested that DiI-C12-labeled domains increased in N immediately after step-shear, while N for DiI-C18-stained membrane transiently decreased. These results demonstrate that membrane microdomains are differentially sensitive to fluid shear stress.

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Acknowledgments

This work was supported by grants to PJB from the National Heart Lung and Blood Institute (R01 HL 07754201-A1) and the National Science Foundation (BES 0238910).

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  1. Department of Bioengineering, The Pennsylvania State University, 205 Hallowell Building, University Park, PA, 16802, USA

    Tristan Tabouillot, Hari S. Muddana & Peter J. Butler

  2. Department of Bioengineering, The Pennsylvania State University, 230 Hallowell Building, University Park, PA, 16802, USA

    Peter J. Butler

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  1. Tristan Tabouillot
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Associate Editor Edward Guo oversaw the review of this article.

Tristan Tabouillot and Hari S. Muddana contributed equally to this work.

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Tabouillot, T., Muddana, H.S. & Butler, P.J. Endothelial Cell Membrane Sensitivity to Shear Stress is Lipid Domain Dependent. Cel. Mol. Bioeng. 4, 169–181 (2011). https://doi.org/10.1007/s12195-010-0136-9

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