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Elevated Level of Fibrinogen Increases Caveolae Formation; Role of Matrix Metalloproteinase-9

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Abstract

The role of the inflammatory agent fibrinogen (Fg) in increased pial venular permeability has been shown previously. It was suggested that an activation of matrix metalloproteinase-9 (MMP-9) is involved in Fg-induced enhanced transcytosis through endothelial cells (ECs). However, direct link between Fg, caveolae formation, and MMP-9 activity has never been shown. We hypothesized that at an elevated level, Fg enhances formation of functional caveolae through activation of MMP-9. Male wild-type (WT, C57BL/6J) or MMP-9 gene knockout (MMP9−/−) mice were infused with Fg (4 mg/ml, final blood concentration) or equal volume of phosphate buffered saline (PBS). After 2 h, mice were sacrificed and brains were collected for immunohistochemical analyses. Mouse brain ECs were treated with 4 mg/ml of Fg or PBS in the presence or absence of MMP-9 activity inhibitor, tissue inhibitor of metalloproteinases-4 (TIMP-4, 12 ng/ml). Formation of functional caveolae was assessed by confocal microscopy. Fg-induced increased formation of caveolae, which was defined by an increased co-localization of caveolin-1 (Cav-1) and plasmalemmal vesicle-associated protein-1 and was associated with an increased phosphorylation of Cav-1, was ameliorated in the presence of TIMP-4. These results suggest that at high levels, Fg enhances formation of functional caveolae that may involve Cav-1 signaling and MMP-9 activation.

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Acknowledgments

Grant sponsors: Contract grant sponsor: NIH; Contract Grant Numbers: P30 GM-103507 (to DL), NS-084823 (to DL and SCT), HL-071010 (to SCT), and NS-051568 (to SCT).

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Authors and Affiliations

  1. Department of Physiology and Biophysics, School of Medicine, University of Louisville, Bldg. A, Room 1115, 500 South Preston Street, Louisville, KY, 40202, USA

    Nino Muradashvili, Reeta Tyagi, Suresh C. Tyagi & David Lominadze

  2. Department of Anatomical Sciences and Neurobiology and Kentucky Spinal Cord Injury Research Center, School of Medicine, University of Louisville, Louisville, KY, USA

    Richard L. Benton

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  1. Nino Muradashvili
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  2. Richard L. Benton
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Correspondence to David Lominadze.

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Muradashvili, N., Benton, R.L., Tyagi, R. et al. Elevated Level of Fibrinogen Increases Caveolae Formation; Role of Matrix Metalloproteinase-9. Cell Biochem Biophys 69, 283–294 (2014). https://doi.org/10.1007/s12013-013-9797-z

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