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Endothelial cytoskeletal elements are critical for flow-mediated dilation in human coronary arterioles

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Abstract

Mitochondrial H2O2 contributes to flow-mediated dilation (FMD) in human coronary arterioles (HCA). We examined the hypothesis that the endothelial cytoskeleton plays a critical role in transducing endothelial wall shear stress into a stimulus for releasing mitochondrial ROS. Phallacidin together with α-, β-tubulin antibodies and Mito-Tracker Red showed the proximity of F-actin, microtubules and mitochondria in endothelial cells. Cytochalasin D (CytoD) and nocodazole (Noc) disrupted endothelial F-actin and microtubules in HCA, respectively, concurrent with a reduction in the generation of cytosolic O2 •− and H2O2 (hydroethidine and dichlorodihydrofluorescein fluorescence) and mitochondrial superoxide (mitoSox) during flow (control: 3.5 ± 1.6, Cyto D: 0.51 ± 0.2, Noc: 0.81 ± 0.6). FMD, but not the dilation to bradykinin or papaverine, was reduced by Cyto D (26 ± 10% vs. 56 ± 3%) or Noc (26 ± 11% vs. 58 ± 7%). These results suggest that cytoskeletal elements are a critical component of the signaling mechanism linking endothelial shear stress and mitochondrial release of ROS in the human coronary microcirculation.

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Acknowledgments

This work was supported by NIH RO1 HL067968 to YL, P01 HL68769 to DG, and a Veterans Administration Merit Award to DG. The authors wish to thank the Division of Cardiothoracic Surgery at the Medical College of Wisconsin, the Cardiothoracic Surgery division at the Zablocki VA Medical Center in Milwaukee, the Cardiothoracic Surgery Group of Milwaukee, the Cardiovascular Surgery Associates of Milwaukee, the Midwest Heart Surgery Institute, and the Wisconsin Heart Group for providing surgical specimens.

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Authors and Affiliations

  1. Department of Medicine, Medical College of Wisconsin, Milwaukee, WI, USA

    Aaron H. Bubolz, David X. Zhang & David D. Gutterman

  2. The Cardiovascular Center, Milwaukee, WI, USA

    Aaron H. Bubolz, David X. Zhang & David D. Gutterman

  3. The Medical College of Wisconsin, The Veterans Administration Medical Center, 8701 Watertown Plank Road, Milwaukee, WI, 53226, USA

    Aaron H. Bubolz, David X. Zhang & David D. Gutterman

  4. The National Center for Research Resources, National Institutes of Health, Bethesda, MD, USA

    Yanping Liu

  5. The Heart and Vessel Diseases Center, Beijing Friendship Hospital, Capital University of Medical Sciences, Beijing, People’s Republic of China

    Hongwei Li

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  1. Yanping Liu
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Correspondence to David D. Gutterman.

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Liu, Y., Li, H., Bubolz, A.H. et al. Endothelial cytoskeletal elements are critical for flow-mediated dilation in human coronary arterioles. Med Biol Eng Comput 46, 469–478 (2008). https://doi.org/10.1007/s11517-008-0331-1

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